The 6th International Triticeae Symposium (6th ITS)
May 31 – June 5, 2009
International Conference Hall II & III, Clock Tower Centennial Hall, Kyoto University, Kyoto, Japan
Taihachi Kawahara
Chair of Local Organizing Committee, Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University, Japan
The Sixth International Triticeae Symposium (6th ITS) was held at the Clock Tower Centennial Hall of Kyoto University in Kyoto, Japan from 31 May to 5 June 2009. The symposium was organized under the joint auspices of the Local Organizing Committee, International Organizing Committee and the National Institute of Agrobiological Sciences (NIAS), Japan. It was supported by the Kyoto University Foundation and the Japanese Society of Breeding. Total of 118 researchers and students, from 20 countries including Australia, Azerbaijan, Canada, China, Czech Republic, Germany, Georgia, Iran, Italy, Japan, Kazakhstan, Mexico, Poland, Russia, Slovak Republic, Spain, Sweden, Turkey, UK, and USA, participated in this symposium (Fig. 1). There were 48 oral presentations including 8 plenary lectures and 49 posters (Fig. 2). Like its predecessors, the Sixth Symposium brought together researchers and students in many different disciplines who had common interest in the one group of grasses, the Triticeae.
Local Organizing Committee
Taihachi Kawahara (Chair of LOC, PGPI, Kyoto University)
Kazuhiro Sato (Secretary of LOC, RIB, Okayama University)
Tomohiro Ban (Kihara Institute for Biological Research, Yokohama City University)
Katsuyuki Kakeda (Graduate School of Bioresource, Mie University)
Masahiro Hishii (Kihara Institute for Biological Research, Yokohama City University)
Takao Komatsuda (National Institute of Agrobiological Science)
Hideho Miura (Obihiro University of Agriculture and Veterinary Medicine)
Tsuneo Sasanuma (Faculty of Agriculture, Yamagata University)
Shigeo Takumi (Graduate School of Agricultural Science, Kobe University)
Hisashi Tsujimoto (Faculty of Agriculture, Tottori University)
International Organizing Committee
Roland von Bothmer (Chair of IOC, Swedish University of Agricultural Science, Sweden)
Mary Barkworth (Vice-chair of IOC, Intermountain Herbarium, Utah State University, USA)
Bradley Shaun Bushman (USDA-ARS Forage and Range Research Lab., USA)
Vojtech Holubec (Department of Gene Bank, Reserch Institute of Crop Production, Czech Republic)
Taihachi Kawahara (Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University, Japan)
Helmut Knupffer (Leibniz Institute of Plant Genetics and Crop plant Research, Germany)
Kazuhiro Sato (Research Institute for Bioresource, Okayama University, Japan)
National Institute of Agrobiological Sciences (NIAS), Tsukuba, Ibaraki, Japan
Teruo Ishige (President, NIAS)
Takuji Sasaki (Vice-President, NIAS)
Hirokazu Handa (Head, Research Planning Section, NIAS)
Takao Komatsuda (Plant Genome Research Unit, NIAS)
PROGRAM
Sunday, May 31, 2009
16:00-20:00 Registration
16:00-20:00 Poster mounting
18:00-20:00 Welcome Reception
Monday, June 1, 2009
09:30-10:00 Opening Session
Chair: K. Sato
Kawahara T., Chair, Local Organizing Committee (Kyoto University, Japan)
Bothmer R. von, Chair, International Organizing Committee (Swedish University of Agricultural Sciences, Sweden)
Ishige, T. (President, National Institute of Agrobiological Sciences, Japan)
10:00-11:00 Plenary Lectures I
Chair: R. von Bothmer
10:00-10:30 Heslop-Harrison J.S. (Leicester University, UK)
Triticeae cytogenomics: results, implications and applications
10:30-11:00 Endo T. (Kyoto University, Japan)
Dissection of barley genome by the gametocidal system
11:00-11:30 Coffee/Tea Break
11:30-13:00 Systematics and Phylogeny I
Chair: R. von Bothmer
11:30-12:00 Blattner F.R. (Leibniz Institute of Plant Genetics and Crop Research, Germany)
Phylogenetic and population level analyses to understand evolutionary processes in Hordeum (Poaceae)
12:00-12:20 Sun G. (Saint Mary's University, Canada)
Molecular evolution and origin of tetraploid Elymus species
12:20-12:40 Goncharov N.P. (Siberian Branch of the Russian Academy of Sciences, Russia)
Taxomony and molecular phylogeny of natural and artificial wheat species
12:40-13:00 Muramatsu M. (Okayama University, Japan)
Wild Triticeae species indigenous to Japan, their features and results of hybridization studies involving wheat and barley cultivars
13:00-14:30 Lunch & Poster View
14:30-15:30 Plenary Lectures II
Chair: J.S. Heslop-Harrison
14:30-15:00 MacKay M. (Bioversity International, Italy)
A strategy to enhance the effective and efficient conservation and use of ex situ plant genetic resources
15:00-15:30 Sasaki T. (National Institute of Agrobiological Sciences, Japan)
Rice as a model for Triticeae genome analysis
15:30-17:30 Systematics and Phylogeny II
Chair: J.S. Heslop-Harrison
15:30-16:00 Barkworth M. (Utah State University, USA)
Identifying genomic groups in the perennial Triticeae by their morphology
16:00-16:30 Coffee/Tea Break
16:30-16:50 Taketa S. (Okayama University, Japan)
Molecular cytogenetic investigation on the origin of two tetraploid Hordeum species, H. secalinum and H. capense
16:50-17:10 Rahiminejad M.R. (University of Isfahan, Iran)
The relationships among the A genome bearing Triticum species as evidenced by SSRs in Iran
17:10-17:30 Kakeda K. (Mie University, Japan)
Molecular phylogeny of the genus Hordeum using thioredoxin-like gene sequences
17:30-19:30 Poster Session
17:30-19:30 Happy Hour
Tuesday, June 2, 2009
09:30-11:00 Domestication and Evolution I
Chair: V. Holubec
09:30-10:00 Mori N. (Kobe University, Japan)
Genetic diversity, evolution and domestication of wheat and barley in the Fertile Crescent
10:00-10:20 Murai K. (Fukui Prefectural University, Japan)
Adaptation of flowering-time in tetraploid wheat by selection of flowering-time genes under domestication
10:20-10:40 Wang N. (National Institute of Agrobiological Sciences, Japan)
Molecular evolution of cleistogamy in barley
10:40-11:00 Nishida H. (Okayama University, Japan)
Structural variation in 5’ upstream region of photoperiodic response genes, Ppd-A1 and Ppd-B1, in wheat
11:00-11:30 Coffee/Tea Break
11:30-13:00 Domestication and Evolution II
Chair: F.R. Blattner
11:30-12:00 Komatsuda T. (National Institute of Agrobiological Sciences, Japan)
Domestication and spike architecture of barley (Hordeum vulgare)
12:00-12:20 Ball T.B. (Brigham Young University, USA)
Typologic and morphometric analysis of phytoliths produced by wheat and barley
12:20-12:40 Takumi S. (Kobe University, Japan)
Natural variation in morphological traits in central Eurasian wild wheat progenitor Aegilops tauschii Coss.
12:40-13:00 Saisho D. (Okayama University, Japan)
Evolutionary process of six-rowed spike in domesticated barley
13:00-14:30 Lunch & Poster View
14:30-16:00 Biodiversity and Genetic Resources I
Chair: M. Barkworth
14:30-15:00 Knüpffer H. (Leibniz Institute of Plant Genetics and Crop Plant Research, Germany)
Genetic resources of Triticeae - cultivated species and genebank collections
15:00-15:20 Holubec V. (Crop Research Institute, Czech Republic)
Annual Triticeae genebank collection
15:20-15:40 Tomita M. (Tottori University, Japan)
Quantitative variation of Revolver transposon-like gene in synthetic wheat and its structural relationship with LARD element
15:40-16:00 Saeidi H. (University of Isfahan, Iran)
Biodiversity of the D-genome species Aegilops tauschii in Iran
16:00-16:30 Coffee/Tea Break
16:30-18:00 Biodiversity and Genetic Resources II
Chair: T. Schnurbusch
16:30-17:00 Yen C. (Sichuan Agricultural University, China)
The tribe Triticeae Dumort. (Poaceae)
17:00-17:20 Kishii M. (Yokohama City University, Japan)
Synthetic wheat production for wheat breeding
17:20-17:40 Garg M. (Tottori University, Japan)
Exploration of Triticeae resource for wheat end product quality improvement
17:40-18:00 Cheng J. (Guizhou University, China)
Natural variation in grain selenium concentration derived from Israeli wild barley, Hordeum spontaneum
18:00-18:20 Yanagisawa T. (National Agricultural Research Center for Western Region, Japan)
Recent breeding objectives of hulled and hull-less barley for food in Japan
Wednesday, June 3, 2009
09:30-20:30 Symposium Excursion
Thursday, June 4, 2009
09:30-11:00 Genomics and Breeding I
Chair: G. Muehlbauer
09:30-10:00 Colmer T.D. (The University of Western Australia, Australia)
Waterlogging and salinity tolerance in wild Hordeum species: physiological basis and prospects for use in wheat improvement
10:00-10:20 Subbarao G.V. (Japan International Research Center for Agricultural Sciences, Japan)
Biological nitrification inhibition (BNI) potential in Triticeae
10:20-10:40 Schnurbusch T. (Leibniz Institute of Plant Genetics and Crop Plant Research, Germany)
Reduced transcript levels at the Bot3 locus in barley (Hordeum vulgare L.) confer increased tolerance to high boron supply
10:40-11:00 Chen G. (National Institute of Agrobiological Sciences, Japan)
Genetic targeting of drought sensitive gene eibi1 of wild barley (Hordeum spontaneum)
11:00-11:30 Cof
11:30-13:00 Genomics and Breeding II
Chair: S.R. Larson
11:30-12:00 Kumlehn J. (Leibniz Institute of Plant Genetics and Crop Plant Research, Germany)
Genetic engineering in cereals: Current technologies for the elucidation of gene functions
12:00-12:20 Sato K. (Okayama University, Japan)
Map based cloning of dormancy QTL in barley
12:20-12:40 Dou Q.W. (North West Plateau Institute of Biology, China)
Diversity of Triticeae as forage crops
12:40-13:00 Konovalov F. (Vavilov Institute of General Genetics, Russia)
Revealing genetic diversity in closely related A-genome diploid wheat species (Triticum boeoticum, T. monococcum, T. urartu) by retrotransposon display
13:00-14:30 Lunch
13:00-14:30 Poster View
14:30-16:00 Genomics and Breeding III
Chair: T.D. Colmer,
14:30-15:00 Sreenivasulu N. (Leibniz Institute of Plant Genetics and Crop Plant Research, Germany)
Revealing seed storage metabolism from genetical genomic and systems biology approaches
15:00-15:20 Turuspekov Y. (Montana State University, USA)
Hardness locus sequence variation in association with grain quality in spring barley (Hordeum vulgare L.)
15:20-15:40 Tsujimoto H. (Tottori University, Japan)
Mutual interchange of genetic variation and genomic information between wild and cultivated species in Triticeae
15:40-16:00 Prieto P. (Consejo Superior de Investigaciones Científicas, Spain)
Development and cytogenetic analysis of Hordeum chilense chromosome 4 introgression lines into durum wheat
16:00-16:30 Coffee/Tea Break
16:30-17:40 Genomics and Breeding IV
Chair: J. Kumlehn
16:30-17:00 Matsumoto T. (National Institute of Agrobiological Sciences, Japan)
Transcriptional landscape of malting barley, Haruna Nijo - using a custom-made oligo array from cDNA sequences
17:00-17:20 Shavrukov Y. (University of Adelaide, Australia)
Salinity tolerance and sodium exclusion in genus Triticum
17:20-17:40 Malik A.I. (The University of Western Australia, Australia)
Submergence tolerance in Hordeum marinum
18:30-21:00 Symposium Dinner
Friday, June 5, 2009
09:30-12:00 Plenary lectures III
Chair: H. Knüpffer
09:30-10:00 Larson S.R. (Utah State University, USA)
Gene, genomic, and trait discovery research in perennial Triticeae grasses
10:00-10:30 Tsunewaki K. (Kyoto University, Japan)
Plasmon analysis in wheat
10:30-11:00 Coffee/Tea Break
11:00-11:30 Muehlbauer G. (University of Minnesota, USA)
The barley coordinated agricultural project (CAP): integrating genomics with breeding
11:30-12:00 Takeda K. (Okayama University, Japan)
Features of East Asian barley and their genetic analyses
12:00-12:30 Business session & Closing remark
Poster List
1. Bushman BS, Barkworth ME. DNA markers: another tool in the toolbox
2. Jakob SS, Blattner FR. Phylogenetic and phylogeographic analyses of Hordeum murinum (Poaceae)
3. Gerus DE, Agafonov AV. Levels of study of StH-genomic Elymus species of Asian Russia and North-Eastern Kazakhstan in connection with a problem of “species-phantoms”
4. Hasheminejad N, Saeidi H, Yoosofi M, Rahiminejad MR. Taxonomy and inter-specific relationships of Agropyron Grant. in Iran
5. Zhang H-Q, Fan X, Huang Y, Sha L-N, Zhou Y-H. Genome constitution of Hystrix komarovii (Poaceae: Triticeae)
6. Ohta S, Fujita Y, Maesaka Y, Hattori M, Iwasaki R. A biosystematic study in Aegilops neglecta – Ae. columnaris species complex
7. Rollo J, Jacobs SWL, Rashid A, Barkworth, ME. Using discriminant analysis to identify genomic groups within the perennial Triticeae
8. Knüpffer H. Geographical distribution patterns of morphological characters in cultivated barley (Hordeum vulgare L.) inferred from botanical varieties
9. Mori N, Watatani H, Ishii T, Kondo Y, Kawahara T, Nakamura C. Intraspecific variation of chloroplast DNA in Aegilops speltoides
10. Ohta A, Kawahara T, Yamane K. Morphological variations of spike and the geographical distribution of subsection Emarginata species, genus Triticum-Aegilops, close wild relatives of wheat
11. Pourkheirandish M, Komatsuda T. The regulatory network underlying the six-rowed spike in barley
12. Sakuma S, Pourkheirandish M, Matsumoto T, Koba T, Komatsuda T. The barley vrs1 gene evolved from duplication of a well-conserved HD-Zip I-class homeobox gene in the Poaceae
13. Morihiro H, Takumi S. Intraspecific variation in leaf shape-related traits in a wild einkorn wheat species Triticum urartu Thum.
14. Tanno K, Bothmer von R, Yamane K, Takeda K, Komatsuda T. Allopolyploidy of the Hordeum murinum complex indicated by a nucleotide sequence of cMWG699
15. Turuspekov Y, Abugalieva S. The variation of SSR profiles in wild and cultivated barley
16. Aliyeva AJ, Aminov NKH. A novel source of germplasm for the development of branched ear wheat
17. An X, Wang D, Yan Y. Isolation and molecular characterization of three novel HMW glutenin subunits from Aegilops tauschii
18. Taguchi J, Kiribuchi-Otobe C, Matsunaka H, Ban T. Analysis grain characteristics of tetraploid wheat gene pool to diversify genetic background of durum wheat
19. Bordbar F, Rahiminejad MR, Saeidi H, Blattner FR. Study of diversity and relationships of the D genome species of Aegilops–Triticum from Iran
20. Gregová E, Medvecká E, Šramková Z, Mihálik D. Estimation of quality of Triticum durum Desf. wheat on the basis of gliadin and glutenin characterisation
21. Mihálik D, Šramková Z, Medvecká E, Horevaj V, Šliková S. Genetic variability in bread wheat (Triticum aestivum L.) of Slovakia based on polymorphism for high molecular weight glutenin subunits
22. Miyazaki T, Ban T. Multiplex Quantitative analysis for trichothecene genes expression of Fusarium graminearum causing head blight on wheat spikes
23. Igartua E, Molina-Cano JL, Gracia MP, Casas AM, Moralejo M, Ciudad FJ, Lasa JM. New tools for the accessibility of the Spanish barley core collection
24. Mosulishvili M, Maisaia I, Shanshiashvili T, Akhalkatsi M. Triticum species in Georgia: diversity, conservation, and taxa of special interest
25. Geraybeyova N, Sadiqov H, Rahimova O, Sadiqova S, Karimov A, Mammadova N, Babayeva S, Abbasov M. Assessment of genetic diversity among Azerbaijan barley genotypes (H. vulgare L.) based on Hordein alleles
26. Sharma S, Röder MS. Study of sequence polymorphism and genetic diversity of sucrose-phosphate synthase genes in bread wheat and its A, B and D genome progenitors
27. Šliková S, Šramková Z, Gregová E, Mihálik D. Composition of high-molecular-weight glutenin subunits in European wheats
28. Zaharieva M, Dreisigacker S, Crossa J, Payne T, Misra S, Hanchinal RR, Mujahid MY, Trethowan R. Genetic diversity within Triticum turgidum L. subsp. dicoccon (Schrank) Thell. (cultivated emmer) and its utilization in wheat breeding
29. Abugalieva S, Abugalieva A, Quarrie S, Turuspekov Y. Identification and mapping of QTLs for grain protein content in common wheat
30. Aydin Y, Cabuk E, Mert Z, Akan K, Bolat N, Cakmak M, Uncuoglu AA. Investigations on yellow rust disease resistance by useful genes and markers in gene-rich regions on wheat chromosomes
31. Bińka-Wyrwa A, Orczyk W, Nadolska-Orczyk A. Regulation of transformation efficiency in polyploid cereals by type and number of selection cassettes
32. Buwan R, Takahashi H, Kato K, Sato Y-i, Komatsuda T, Nakamura I. Sequence variation of the 20th exon within PolA1 gene among Triticeae species
33. Cagirgan AMI, Ullrich SE, Ozbas MO. High frequency and a wide spectrum of mutations in ‘BARONESSE’ barley fields
34. Miroshnichenko D, Poroshin G, Dolgov S. Characterization of growth and yield of transgenic wheat plants overexpressing vacuolar Na+/H+ antiporter genes
35. Kikuchi R, Kawahigashi H, Ando T, Tonooka T, Handa H. The flowering pathway under short day in barley
36. Martín AC. The potential of Hordeum chilense cytoplasm in the development of CMS systems in Triticeae crops
37. Martinek P, Dobrovolskaya O, Röder MS, Börner A. Agronomic traits and genetic determination of winter wheat lines (Triticum aestivum L.) with multirow spike
38. Martinek P. Breeding Triticale (X Triticosecale Wittmack) for improved breadmaking quality
39. Miroshnichenko D, Poroshin G, Dolgov S. Gene flow from genetically modified to cultivated wheat plants
40. Mishina K, Manickavelu A, Sato H, Katsumata M, Sassa H, Koba T. Observation of pollen tube growth and molecular mapping of Kr genes in common wheat-rye hybridization
41. Nadolska-Orczyk A, Zalewski W, Galuszka P, Orczyk W. Posttranscriptional silencing of CKX genes, regulating cytokinin level in barley by RNA interference
42. Niwa S, Kikuchi R, Handa H, Ban T. Genetic variability of MRP gene constituting ‘Qfhs.kibr-2DS’ QTL to reduce Fusarium mycotoxin accumulation among hexaploid wheats
43. Tanaka H, Arakawa T, Tsujimoto H. Alien glutenin subunits expressed in common wheat endosperm affect on the composition
44. Tonooka T, Aoki E, Yoshioka T, Taketa S, Kiribuchi-Otobe C. Characterization of a β-glucanless mutant in barley
45. Kidou S, Yokota S, Yoshida K. Virus-induced gene silencing of P23k in barley leaf reveals morphological changes involved in secondary wall formation
46. Zhang LQ, Liu DC, Zheng YL, Yen Y. Spontaneous amphidiploidization via unreduced gametes is a universal phenomenon for Triticum turgidum - Aegilops tauschii hybrids
47. Mizuno N, Yamasaki M, Matsuoka Y, Kawahara T, Takumi S. Population structure of central Eurasian wild wheat progenitor Aegilops tauschii Coss.
48. XF Zhang, DC Liu, WL Yang, JZ Sun, DW Wang, HQ Ling and AM Zhang. Molecular markers for systematic characterization of low molecular weight glutenin subunits in common wheat (Triticum aestivum L.)
49. Nishinaka M, Okumoto Y, Kato K, Kawahara T, Tanisaka T. Genetic diversity of high molecular weight glutenine subunits in wheat landraces
ABSTRACTS & TITLES
1. Oral Presentation
1-1.
Triticeae cytogenomics: results, implications and applications
Heslop-Harrison JS (Pat)1, Contento A1, Graybosch RA2, Ali N1, Kuhn G CS1, Saeidi H1,3, Kalpande H1,4, Schwarzacher T1
1Department of Biology, University of Leicester, LE1 7RH, UK
2USDA-ARS, University of Nebraska, Lincoln, NE 68583, USA
3Faculty of Science, University of Isfahan, Isfahan, Iran
4Marathwada Agriculture University, Maharashtra, India
The study of the genomics of the Triticeae has been a story of continuous progress over nearly a century. After the definition of species and polyploids, the seminal work of Kihara and Sears showed the basic number of x=7 and developed the concepts of the genome. Following these studies, relationships and the genetics of characters including disease resistance and quantitative traits were investigated, where aneuploids and synthetic hybrids were invaluable, leading up to our current state of knowledge about genes and DNA sequences. These academic advances have been paralleled by exploitation of the knowledge in plant breeding. In this talk, I will discuss our cytogenomics research in the Triticeae, studying genome evolution at a large scale. Tandemly repeated satellite DNA sequence arrays that make a substantial parts of Triticeae genomes, and include some of the most conserved sequence motifs as well as abundant variable sequences which are not conserved even between closely related genomes. I will report our increasing understanding of the chromatin code – the changes in methylation and histones which are responsible for epigenetic effects, and may play an important role in nuclear architecture and genome interactions in polyploids. Finally, I will discuss the applications of this work in the use of hybrids to interchange genetic variation and widen the genepool available to plant breeders. Related publications and information are available from www.molcyt.com.
1-2.
Dissection of barley genome by the gametocidal system
Endo TR
Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Barley is one of the major cereals in the world. The analysis of genomes is important in modern breeding, but the barley genome is too huge and complicated that it is still difficult to arrange sequenced pieces of the genome in order. It is desirable to have the genome divided into small pieces in separate plant lines. Each of the barley chromosomes has been added to common wheat and such barley chromosome addition lines are useful in locating the positions of genes and DNA markers on a chromosome. It has been proved that the added barley chromosomes in common wheat can be fragmented genetically by the gametocidal system. This system involves unique chromosomes, called gametocidal (Gc) chromosomes, that were derived from some wild species of the genus Aegilops related to common wheat. When the Gc chromosome exist in common wheat in monosomic condition, two types of gamete are produced, one carrying the Gc chromosome, the other without the Gc chromosome, and chromosome breakage occurs only in the latter gamete. Such Gc-induced chromosomal breakage leads to either the sterility of gametes or the production of fertile gametes carrying chromosomal mutations. Although no molecular mechanism of the gametocidal system is known, this system has been applied successfully to the generation of common wheat lines carrying segments of barley chromosomes. Thus, it has become possible to dissect the barley genome. In this talk I will describe the progress in the dissection of the barley genome.
1-3.
Phylogenetic and population level analyses to understand evolutionary processes in Hordeum (Poaceae)
Blattner FR
Leibniz Institute of Plant Genetics and Crop Research (IPK), D-06466 Gatersleben, Germany
Hordeum consists of 31 species, distributed in temperate and arid regions of the Northern Hemisphere, South Africa and southern South America. About 50% of the species thrive in this latter region, making it the diversity center of the genus. To analyze evolutionary processes in the genus and to explain the high species number in the Americas, phylogenetic and phylogeographic analyses together with molecular dating approaches, reconstruction of historical biogeography, and ecological niche modeling were conducted for species and species groups of the genus.
The genus originated about 12 million years (My) ago in Western Asia and colonized its extant distribution area by at least seven intercontinental long-distance dispersals. About 4 My ago it reached South America, where a rapid radiation took place during the last 2 My. While high speciation rates are characteristic for the Americas, Eurasian Hordeum lost most of its genetic and species diversity through extinction during the Pleistocene. Eurasian species were mostly restricted to small refugial populations during ice-age cold cycles, while particularly in southernmost Patagonia large populations survived the ice-age without spatial or genetic restrictions. Therefore, speciation in Eurasia involved mostly severe genetic bottlenecks, while South America was characterized by vicariance-driven speciation. These differences are clearly reflected in chloroplast diversity, resulting in specific patterns in the Old and New World.
1-4.
Molecular evolution and origin of tetraploid Elymus species
Sun G
Biology Department, Saint Mary's University, Halifax, NS, B3H 3C3 Canada
It is well known that Elymus arose through hybridization between representatives of different genera and several different polyhaplomic genomes have been described. Cytogenetically, five basic genomes (St, H, Y, P, and W) in different combinations have been found in the genus. The vast majority of species are tetraploids and they are characterized by having the StY genome or the StH genome. It is not known where the Y genome originated, although it is a common in Elymus from Central and East Asia. Phylogeny and origin of tetraploid Elymus is far from clear. It has been hypothesized from isozymic and cytological studies of Elymus species that the Old and New World taxa may be of separate origin of the H genome in the StH genome species. To test this hypothesis, and estimate the phylogenetic relationship of polyploid Elymus species within the Triticeae, single copy of nuclear gene, the second largest subunit of RNA polymerase II (RPB2), was analyzed with Elymus species containing StH or StY genomes and diploid species. Our data indicated that the Eurasian and American StH genome species have independent alloploid origins with different H-genome donors, and provides some insight on the origin of Y genome and its relationship to other genomes in Elymus.
1-5.
Taxonomy and molecular phylogeny of natural and artificial wheat species
Goncharov NP, Golovnina KA, Kondratenko EYA
Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 10 Lavrentyev ave., Novosibirsk 630090, Russia
The effective use of wheat genetic resources is connected with their conservation strategy and mechanism of their utilization. Producing of wheat amphiploids with genomes of related species is an effective way for this purpose. The problem of such amphiploid preservation has been under investigation during last hundred years. Artificial amphiploids should be given names and places in the genus Triticum taxonomy for preservation in genebanks. In this connection the carefulness in working with genus Triticum classification is very important not only to find the solution of problems in wheat origin and its phylogeny, but also to collect and estimate wheat biodiversity preservation. The inheritance of domesticated and taxonomically important characters has been studied in present work. An attempt to integrate the results of different comparative-genetic analyses of wheats and their molecular taxonomy has been made. The correspondence of earlier evolutionary specifications to the phylogenetic relationships within the genus Triticum species has been estimated using chloroplast and nuclear DNA sequence data. The results of the provided molecular analysis indicated close relationship of all hexaploid species. Three different variants of investigated genes have been detected in diploid A genome Triticum. The detailed analysis showed that one of these variants was a progenitor for all A genomes of all polyploid Triticum species except hexaploid Timopheevii group species and the artificial ones.
Presence of competing wheat classifications and using illegitimate species names are causing confusion in different groups of research community. The possibility of using different classifications of genus Tritucum L. for molecular-biological, genetic and phylogenetic investigations, for collecting and identifying wheat accessions and breeding practice is discussed.
1-6.
Wild Triticeae species indigenous to Japan, their features and results of hybridization studies involving wheat and barley cultivars
Muramatsu M
Ezucho 3-6, Okayama 700-0028, Japan
Studies on the wild Triticeae species indigenous to Japan have revealed some biological phenomena; the main results are summarized here: (1) Observations made for many years indicate that three species, Elymus tsukushiensis, E. humidus, and E. ciliaris are especially tolerant of the humid monsoon climate. (2) From the cross pollination of these three species with species of Triticum, Aegilops, Secale and Hordeum, it was possible to obtain F1s by using embryo rescue. In crosses of E. tsukushiensis and E. humidus as female by Hordeum, hybrid plants had many sectors and polyhaploid plants of the female parents often resulted. In the combination involving Triticum, an amphiploid line was produced with colchicine treatment when E. ciliaris was the parent, whereas with E. tsukushiensis it was not possible to obtain amphiploids in spite of repeated treatments. The same behavior occurred in hybrids E. tsukusiensis x rye. Attempts to double the chromosome number of E. tsukushiensis and E. ciliaris have never succeeded. It appears that the formation of polyploids may be under genetic control. The term, “definitomodis genetic conditions” is proposed to designate such control. The term was suggested by the late Dr. G. Redei. (3) For the number of spikelets per spike node, which is 1~2 in Elymus and 1 in Triticum, the genes involved may be on the chromosome of the homoeologous group 2. Nullisomics of that group in hexaploid wheat cv. Chinese Spring often have two spikelets resembling Elymus, indicating dosage effect of the duplicated genes; that is, the decrease from six to four doses may induce the Elymus-like spikelet phenotype.
1-7.
A strategy to enhance the effective and efficient conservation and use of ex situ plant genetic resources
Mackay MC1, Guarino L2, Street KA3
1Bioversity International, Via dei Tre Denari 472/a, 00057 Maccarese, Rome, Italy
2Global Crop Diversity Trust, C/o FAO, Viale delle Terme di Caracalla, 00153 Rome, Italy
3International Centre for Agricultural Research in the Dry Areas (ICARDA), P.O. Box 5466, Aleppo, Syrian Arab Republic
The conservation and use of plant genetic resources (PGR) has a history dating back to the first domestication of plants by humans. In the 20th Century significant efforts were made to collect and conserve these resources in ex situ genebanks. This was done initially at an institutional level but by the end of the Century there was a whole range of conservation and use paradigms, including private, state, national, regional and international collections. Each genebank had its own information management system, mandate, standards and modus operandi. Despite this diverse array of arrangements for conserving PGR, they still need to be studied and properly documented to ensure their effective use. To facilitate this on a global level standards are required: standards of data quality and quantity for documentation; standards to uniquely identifying genotypes; standards to identify duplication; standards for aggregating information across genebanks. The ongoing development and deployment of such standards is leading PGR towards a workable global structure that will provide access to, and practical ways to identify and use, the millions of accessions stored in the world’s genebanks. This paper will describe how the jigsaw puzzle of this global system is being put together.
1-8.
Rice as a model for Triticeae genome analysis
Sasaki T, Matsumoto T
National Institute of Agrobiological Sciences, 1-2, Kannondai 2-chome, Tsukuba, Japan
Rice genome research has generated major advances in plant science including a wide array of genetic resources and genomic information. Utilization of the rice genome sequence information is now a two-faceted strategy aimed at increasing world food production. The first target focuses on complete understanding of rice biology based on the genome composition and function to elucidate the components involved in grain yield, resistance to biotic/abiotic stress, and adaptability to extreme environmental conditions. The second target focuses on using the rice genome as a model for understanding other cereals crops through comparative approaches. The completion of the rice genome sequence in 2004 led to elucidation and isolation of many important genes that support rice molecular breeding and functional characterization of genes from other cereal species. Rice, and the Triticeae diverged from a common ancestor about 40 MY ago. Both genomes however retain the same order of genes in corresponding genomic blocks. This synteny between rice and the Triticeae has been clarified by cross-genetic mapping and further indicates conservation of gene order across regions of the chromosomes. In some cases, rice gene homologs could be identified based on synteny irrespective of its function in Triticeae. The syntenic relationships in cereal crops extend to the sequence level as revealed by comparative analysis of specific regions including the regulatory sequence of corresponding genes. Although the genome size of Triticeae is several times larger than rice due to high-degree of insertion of transposable elements, macrocolinearity and microcolinearity in both genomes must be exploited to accelerate genome analysis. The high-quality rice genome sequence and subsequent achievements in rice genomics should provide a model towards a comprehensive characterization of Triticeae genome structure and function. The availability of Triticeae genome information will provide additional tool for crop improvement to ensure a stable food supply.
1-9.
Identifying genomic groups in the perennial Triticeae by their morphology
Rollo J1, Jacobs SWL2, Rashid A3, Barkworth ME1
1Intermountain Herbarium, Dept. of Biology, Utah State University, Logan, Utah, 84322-5305, USA
2National Herbarium of New South Wales, Mrs Macquaries Road, Sydney, New South Wales, 2000, Australia
3University of Peshawar Botanic Garden, University of Peshawar, Peshawar, Northwest Frontier Province, Pakistan
Our goal was to determine whether morphology could be used to predict the genomic constitution of perennial Triticeae with solitary spikelets. We scored 77 characters (58 quantitative, 19 qualitative) on 219 specimens. The specimens represented 78 taxa and 13 different genomic groups. Discriminant analysis of the quantitative characters indicated that it is possible to use morphology to place perennial Triticeae with solitary spikelets in their genomic group. Which characters are most important depends on the sample size of the groups in the analyses. The qualitative characters proved to be of little value. The data have been used to initiate development of an online, multi-access key to the groups and to aid in development of a dichotomous key to the groups based on our findings. These have been tested on specimens that were not included in the analyses.
1-10.
Molecular cytogenetic investigation on the origin of two tetraploid Hordeum species, H. secalinum and H. capense
Taketa S1, Nakauchi Y2, Bothmer von R3
1Research Institute for Bioresources, Okayama University, Kurashiki 710-0046, Japan
2Faculty of Agriculture, Kagawa University, Miki 761-0795, Japan
3Department of Crop Science, Swedish University of Agricultural Sciences, SE-230 53 Alnarp, Sweden
We previously reported that the two tetraploid Hordeum species, H. secalinum and H. capense are allotetraploids carrying the Xa genome of H. marinum and the I genome of an unidentified diploid species (Taketa et al. Hereditas 130: 185-188, 1999). In the present study, intraspecific variation in each tetraploid species was investigated with regard to intergenomic translocations and chromosomal distribution of rDNA sites. Genomic in situ hybridization revealed that three H. secalinum accessions examined did not carry intergenomic translocations, but that two of three H. capsense accessions carried a pair of intergenomic translocations. Multicolour fluorescent in situ hybridization was applied to analyse chromosomal distribution of rDNA sites. In H. secalinum, two rDNA patterns were found and they differed in the presence or absence of an additional 5S rDNA site in the long arm of a submetacentric chromosome of the Xa-genome origin. The additional 5S rDNA site was also found in all H. capense accessions examined. The additional 5S rDNA site is characteristic of H. marinum subsp. gussoneanum, but this site is absent in H. marinum subsp. marinum. Polymorphisms in 5S rDNA site infer that H. secalinum has two lineages, one having subsp. gussonuanum and the other having subsp. marinum, as the Xa-genome donor. We suppose that H. capense is originated from a limited number of H. secalinum accessions that were introduced through migration of European people to South Africa.
1-11.
The relationships among the A genome bearing Triticum species as evidenced by SSRs in Iran
Ehtemam MH1, Rahiminejad MR2, Saeidi H2, Sayed Tabatabaei BE1, Krattinger S3, Keller B3
1Department of Agriculture, Isfahan University of Technology, Isfahan, 84156-83111, Iran
2Department of Biology, University of Isfahan, Iran
3Institute of Plant Biology, University of Zurich, Switzerland
Microsatellites are polymorphic, multi-allelic and co-dominantly inherited molecular markers, which have become the marker of choice in plant genetics and breeding studies. Genetic relationships of 55 genotypes belonging to 8 Triticum of A genome bearing species (T. monococcum, T. boeoticum, T. urartu, T. durum, T. turgidum, T. dicoccum, T. dicocoides and T. aestivum) collected from Iran and some accessions from other areas were examined using SSR markers. Thirty-two polymorphic primers were used to detect the biodiversity and relationships among the A genome bearing species. A total of 411 alleles were revealed from which 349 were polymorphic. Among the species studied T. durum showed the maximum number of polymorphic loci (186) and T. dicocoides the minimum (31). The observed (HO) and expected heterozygosity (HE) varied from 0 to 0.98 (average of 0.49) and 0 to 0.92 (average of 0.79), respectively. UPGMA cluster analysis based on Nei 1972 coefficient classified the genotypes examined into three main groups which were corresponded to three ploidy levels. Analysis of Molecular Variance (AMOVA) results indicated that most of the genetic variance occurred among populations between the assumed groups, although there was significant variance within the populations (75.5%). A significant correlation was not detected between the poloidy levels of the species studied (p < 0.05). Results of analysis showed that lower genetic distance was observed in diploid groups (0.1) comparing with tetra- and hexaploids populations (0.14-0.36). T. urartu and T. turgidum showed the highest genetic distance (0.36) as similar as genetic distance of T. dicoccum and T. aestivum (0.36). The results of AMOVA analysis were in accordance with cluster analysis and showed that genotypes were not differentiated based on their geographical regions.
1-12.
Molecular phylogeny of the genus Hordeum using thioredoxin-like gene sequences
Kakeda K1, Taketa S2, Komatsuda T3
1Graduate School of Bioresources, Mie Univ., Tsu514-8507, Japan
2Research Institute for Bioresources, Okayama Univ., Kurashiki 710-0046, Japan
3National Institute for Agrobiological Sciences, Tsukuba 305-8602, Japan
Phylogenetic relationship in the genus Hordeum was investigated based on nucleotide sequences of the thioredoxin-like (HTL) gene. HTL gene was originally isolated in diploid H. bulbosum as a single copy gene closely linked to the self-incompatibility (S) locus. We amplified PCR fragments homologous to HTL from 11 species including 16 taxa (25 accessions), which cover mainly diploid accessions together with several tetraploid accessions in H. marinum and H. murinum. We determined nucleotide sequences of the variable region (949 to 1270 bp) that mainly includes 5’-UTR and 2 introns. Phylogenetic analysis based on these sequence data clearly demonstrated a divergence of 4 basic genomes H (H. vulgare and H. bulbosum), X (H. marinum), Y (H. murinum) and I (other species) in the genus Hordeum. Phylogenetic clustering also inferred 2 clades separating one containing H and Y, and the other containing X and I genomes, although the bootstrap value of the latter was lower than that of the former. In the I genome, 4 American species (H. brachyantherum, H. chilense, H. pubiflorum, H. pusillum) were confirmed to be closely related each other and divergent from Asian species (H. brevisubulatum, H. bogdanii, H. roshevitzii). Two diploid cytotypes of H. marinum (ssp. marinum and ssp. gussoneanum) were suggested to be involved in the formation of the tetraploid cytotype (ssp. gussoneanum). Two tetraploid cytotypes of H. murinum (ssp. murinum and ssp. leporinum) shared 2 distinct sequences, one related to that of the diploid cytotype (ssp. glaucum) and the other unique to them.
1-13.
Allelic diversity at chloroplast microsatellite loci among polyploid wheat species
Mori N
Laboratory of Plant Genetics, Graduate School of Agricultural Science, Kobe University, Kobe, Japan
The domestication of wheat and barley was the most important step in the emergence of farming communities that later led to the ancient civilisations of Mesopotamia. Several lines of evidence indicate that emmer wheat (Triticum turgidum subsp. dicoccum, genome: AABB, 2n = 28) was the earliest domesticated wheat derived from wild emmer (T. turgidum subsp. dicoccoides, genome: AABB) and that the domestication occurred within the Fertile Crescent of southwest Asia. Chloroplast DNA fingerprinting of wild and domesticated emmer wheat revealed that two distinct maternal lineages were involved in their domestication and thus suggested that domestication of emmer wheat occurred independently at least two times. Further survey in common wheat revealed that only one of the two maternal lineages of emmer wheat might have transmitted to common wheat through allopolyploidy evolution.
Based of these results the process and geography of wheat domestication will be discussed.
1-14.
Adaptation of flowering-time in tetraploid wheat by selection of flowering-time genes under domestication
Murai K
Fukui Prefectural University, 4-1-1 Matsuoka-kenjojima, Eiheiji-cho, Fukui 910-1195, Japan
There are three major gens responsible for variation in vernalization requirement in temperate cereals such as wheat and barley, VRN1, VRN2, and VRN3. Vernalization up-regulates VRN1 and VRN3, which activate transition from vegetative to reproductive phase (termed flowering). VRN2 is a repressor of flowering, and down-regulated by vernalization. VRN1 is a homolog of Arabidopsis APETALA1/FRUITFULL MADS-box genes, and VRN2 encodes a putative zinc finger and a CCT domain transcription factor. Genetic and molecular studies suggested that the VRN2 protein suppresses VRN1 expression. VRN3 is a homolog of Arabidopsis FLOWERING LOCUS T, which proteins function as the florigen that moves from the leaves into the shoot apex to determine floral meristem identity, leading to the floral organ formation. We performed expression, mutant and transgenic studies to clarify the genetic network of these flowering-time genes, VRN1, VRN2, and VRN3. Expression analysis of a VRN1 deletion mutant suggested that VRN3 is up-regulated by VRN1. Furthermore, transgenic analysis indicated that VRN3 suppresses VRN2 expression. Based on these results, we recently present the VRN1-VRN3-VRN2 triangle model for the regulation of floral transition in wheat, in which VRN1 is upstream of VRN3 with a positive feedback loop through VRN2. In this paper, I discuss the molecular mechanism of adaptation of flowering-time in tetraploid wheats on the basis of the VRN1-VRN3-VRN2 triangle model. Especially, I would like to focus on selection of flowering-time genes under domestication.
1-15.
Molecular control of cleistogamy in barley
Wang N1, Nair S1, Turuspekov Y1, Pourkheirandish M1, Sinsuwongwat S1, Sameri M1, H2, Honda I3, Watanabe Y3, Stein N4, Wicker T5, Tagiri A1, Nagamura Y1, Matsumoto T1, Komatsuda T1
1National Institute of Agrobiological Sciences, Plant Genome Research Unit, 1-2-1 Kannondai, Tsukuba, Ibaraki 305 8602, Japan
2Institute of the Society for Techno-innovation of Agriculture, Forestry and Fisheries (STAFF), 446-1 Ippaizuka, Kamiyokoba, Tsukuba, Ibaraki, 305-0854, Japan
3National Institute of Crop Science (NICS) Tsukuba, Kannondai 2-1-18, Ibaraki 305-8518, Japan
4Group Genome Diversity, Dep.Genebank Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Corrensstr. 3, D-06466 Gatersleben, Germany
5Institute of Plant Biology, University of Zürich, Zürich CH8008, Switzerland
Open/close flowering is considered to be one of the most important traits for alternative breeding programs in both barley and wheat, such as for increase outcrossing rate or conversely restict gene flow of GM. In open flowers lodicules force leamma and parlea apart by the swelling itself, however, some natural variants that tightly closed throughout the anthesis exist in cultivated barley. In this report, the closed flowering is defined cleistogamy in the strict sense and the isolation of Cly1, which lies in a genomic region of chromosome 2HL, showing some colinearity with a part of rice chromosome 4. The colinearity is interrupted by a micro-inversion. A fine mapping allowed Cly1 to be located within a 0.7cM interval defined by two ESTs, and the locus co-segregated with an ortholog of a rice transcription factor. A barley EST homologous with this ortholog was used to screen a barley BAC library. The development of additional markers narrowed the location of Cly1 to a 7kbp region which contained only a single ORF. The expression of this sequence (presumed to be Cly1) was specific to the lodicule, and transcripts were abundant in the immature spikes. The Cly1 coding sequence of a core collection of 274 cultivars was resequenced, and this produced a clear correlation between cleistogamy and the presence of a synonymous SNP located in a conserved domain within the C-terminal region. In the presence of the recessive allele, transcription was suppressed, suggesting that the development of the lodicule fails when the Cly1 protein accumulates.
1-16.
Structural variation in 5’ upstream region of photoperiodic response genes, Ppd-A1 and Ppd-B1, in wheat
Nishida H, Yoshida T, Akashi Y, Kato K
Graduate School of Natural Science & Technology, Okayama University, 1-1-1, Tsushima-Naka, Kita-ku, Okayama 700-8530, Japan
Three dominant alleles Ppd-A1, Ppd-B1, and Ppd-D1 confer photoperiod-insensitivity, while their recessive alleles confer photoperiod-sensitivity in wheat. To date, no allelic variation have been found to explain the difference between photoperiod-insensitive and -sensitive alleles in Ppd-A1 and Ppd-B1, whereas the large deletion in 5’ upstream region of Ppd-D1 allele, that does not exist in ppd-D1 allele, is considered to be an causal structural variation. In this study, Ppd-1 homoeoalleles from two Japanese cultivars “Chihokukomugi” (genotype: Ppd-A1 ppd-B1 ppd-D1) and “Abukumawase” (genotype: ppd-A1 Ppd-B1 Ppd-D1) were analyzed to identify such the allelic variation. As for Ppd-A1, a 1085 bp deletion was detected in the 5’ upstream region of Ppd-A1 allele, while it was not in ppd-A1 allele. This deletion covers most part of the deletions in Ppd-D1 allele and in tetraploid’s Ppd-A1 allele reported before. As for Ppd-B1, a 308 bp insertion was detected in the 5’ upstream region of Ppd-B1 allele, while it was not in ppd-B1 allele. This insertion site exists within the deletion region of Ppd-A1 and Ppd-D1. Furthermore, the approx. 100 bp region around the insertion site is conserved among species, barley, rice, and Brachypodium, suggesting that this sequence is involved in the control of Ppd-1 expression. In addition, no important allelic variations were detected in the coding region of Ppd-A1 and Ppd-B1. Therefore, it was strongly suggested that these structural variations explain the difference between photoperiod-sensitive and -insensitive alleles.
1-17.
Domestication and spike architecture of barley (Hordeum vulgare)
Komatsuda T1, Sakuma S1,2, Koba T2, Pourkheirandish M1, Matsumoto T1, Gottwald S3, Hensel G3, Kumlehn J3, Stein N3
1National Institute of Agrobiological Sciences (NIAS), Plant Genome Research Unit, Kan-non-dai 2-1-2, Tsukuba, Ibaraki 305 8602, Japan
2Graduate School of Horticulture, Chiba University, Matsudo, Chiba 271-8510, Japan
3Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany
The barley spike is composed of triplets (each with one central and two lateral spikelets) arranged alternately at the rachis nodes. This arrangement of spikelets is common in Hordeum species but unique in Triticeae. All three spikelets of the six-rowed barley cultivars are fully fertile and able to develop into grains, but the lateral spikelets of two-rowed barley are reduced in size and sterile. Wild barley (H. vulgare ssp. spontaneum) is two-rowed and its arrow-like triple spikelets, a product of disarticulation of the rachis, are result of an adaptive specialization that took place under native conditions. Some 10,000 years ago, early farmers generated prototypes of cultivated barley with non-brittle rachis. Later, six-rowed spikes that stably produced three times the usual grain number were produced during domestication. The six-rowed spike 1 (vrs1) gene was isolated by a map-based approach. The vrs1 locus included HvHox1 encoding a homeodomain-leucine zipper I–class protein, a potential transcription factor common in plants. Analysis of a plenty of Swedish mutant lines suggested mutational events at regulatory regions of Vrs1 in addition to a variety of structural changes of encoded polypeptides by codon changes, altered splicing, new stop codons and frame shift. Reverse genetics using TILLING population was successful in the detection of new mutant alleles. In six-rowed barley cultivars, analogous mutational events were detected, which indicate a multiple origin of six-rowed barley. Vrs1 is strongly transcribed in lateral spikelet primordia in the triple mound stage as shown by in situ hybridization analysis. Down-regulation of Vrs1 gene expression by inverted-repeat RNAi in transgenic barley lines provided evidence of the biological function of Vrs1 on the determination of spike row number. Evolutionary patterns of Vrs1 in cereals will be presented.
1-18.
Typologic and morphometric analysis of phytoliths produced by wheat and barley
Ball TB
370 A JSB, Brigham Young University, Provo, UT 84602, USA
Solid deposits of amorphous hydrated silica are formed at specific intracellular and extracellular locations in many plant taxa, including all taxa in Triticeae. These deposits of silica are called phytoliths, literally meaning “plant-rocks”. Many plants produce phytoliths with morphological characteristics unique to the taxon. When plant tissue decomposes, any phytoliths formed are released into the surrounding environment thus becoming microfossils of the plants that produced them. Analysis of microfossil phytoliths can provide information to researchers in a wide variety of disciplines, including, archaeobotany, paleoecology, phytogeography and systematics. This paper reviews current methodologies and results of typologic and morphometric analysis of wheat and barley phytoliths. It further presents paradigms for distinguishing between phytoliths produced by these taxa at the genus and species level.
1-19.
Natural variation in morphological traits in central Eurasian wild wheat progenitor Aegilops tauschii Coss.
Takumi S1, Morihiro H1, Nishioka E1, Kawahara T2, Matsuoka Y3
1Graduate School of Agricultural Science, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan
2Graduate School of Agriculture, Kyoto University, Muko 617-0001, Japan
3Fukui Prefectural University, Matsuoka, Eiheiji, Yoshida, Fukui 910-1195, Japan
Aegilops tauschii Coss. (syn Ae. squarrosa L.) is a wild autogamous diploid wheat species. It has a wide natural species range in central Eurasia, spreading from northern Syria and Turkey to western China. Ae. tauschii is also known as the D genome progenitor of hexaploid bread wheat. The genealogical and geographic structure of variation in morphological traits was analysed for Ae. tauschii using a diverse array of 203 sample accessions that represented the entire species range. Our previous studies reported that significant longitudinal and latitudinal clines were detected for spikelet size, and that there exist significant longitudinal and latitudinal clines in flowering time. Totally 12 traits including anther- and pistil-shape and internode length were used in this study. Large natural variation was found for the all examined traits in the Ae. tauschii accessions. Geographically, significant latitudinal clines were detected for anther size, internode length and spike length. Anther tended to be small in the eastern region. Internode also tended to be short, whereas spike to be long in the eastern region. Based on these results, we discuss the patterns of intraspecific divergence and morphological diversification in the course of Ae. tauschii’s long dispersal from Transcaucasus to China.
1-20.
Evolutionary process of six-rowed spike in domesticated barley
Saisho D1, Pourkheirandish M2, Komatsuda T2
1Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan
2National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba 305-8602, Japan
The origin of six-rowed spike was one of the most seminal evolutionary events in domesticated barley. It has revealed that six-rowed spike morphology was caused by recessive mutation in Vrs1 locus, which encoded homeodomain-lecine zipper I homeobox gene (HvHox1). In order to investigate the evolutionary process of the six-rowed barley, we performed comprehensive molecular polymorphic analysis using wild and domesticated barley accessions collected from all over the world. Approximately 2 kb spanning the entire region of HvHox1 gene was sequenced in 136 wild barley core-collection distributed from ICARDA, 267 world representative collection of domesticated barley from RIB, Okayama University, 41 North African accessions from NIAS and 13 Western Asian accessions from USDA. Of the 224 six-rowed types found in our sample panel, we identified four types of vrs1 alleles, designed to vrs1.a1, vrs1.a2, vrs1.a3 and vrs1.a5. The frequencies of each allele in this panel were 72%, 10%, 8% and 9%, respectively. A haplotype genealogy was constructed using the diallelic substitution nucleotide polymorphisms observed in HvHox1 gene. Each of the four types of vrs1 alleles was assigned to significantly separate lineages on this phylogenic tree. The assessments of geographic distribution of each vrs1 alleles showed the contrasting patterns among the alleles. The two alleles (vrs1.a1 and vrs1.a3) were spread to the worldwide range from Europe to East Asia. In contrast, the residual two alleles were locally distributed to either ‘Occidental’ regions (vrs1.a2) or ‘Oriental’ regions (vrs1.a5). These evidence indicated that the recessive vrs1 mutation events responsible for six-rowed barley were repeatedly occurred in process of the barley domestication and that the repetition of the migrations to westward and / or to eastward in the Old World could generate the geographic distribution patterns of vrs1 alleles revealed in this study.
1-21.
Genetic Resources of Triticeae - cultivated species and genebank collections
Knüpffer H
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany
Of the ca. 350 species and ca. 30 genera estimated for the Triticeae, 111 species of 19 genera are either cultivated or useful wild species. An overview is given of these species and their main uses. More than 1,250,000 accessions of Triticeae are maintained in the world’s genebanks, thus comprising one-fifth of the estimated world germplasm holdings. Based on a survey of online information resources, Triticeae accessions belonging to 35 genera (among them 12 hybrid genera) and almost 300 species are documented to occur in almost 300 genebank collections. Summaries of the world holdings per genus, species, regions where the genebanks are located, and the largest collections of the major genera are provided. For the larger genera (in terms of total number of genebank accessions), i.e., Triticum, Hordeum, xTriticosecale, Aegilops, Secale, Elymus and Agropyron, the worldwide germplasm collections are surveyed with more detail. Existing international or regional cooperation programmes, germplasm databases and cultivar registers with pedigree information for these genera are briefly described. For the major cereal crops, core collections and genetic stocks collections are also mentioned. Due to its growing importance as a model plant in genomics research, the genus Brachypodium closely related to the Triticeae is also included in the surveys. The presentation aims at providing background information for plant breeders and crop plant researchers about the germplasm available in ex situ genebank collections, to make this wealth of material more easily accessible.
1-22.
Annual wild Triticeae Gene Bank collection
Holubec V
Dept. of Gene Bank, Crop Research Institute, Drnovska 507, 16106 Praha 6 Ruzyne, Czech Republic
Gene bankers have a greater responsibility for material than bankers have for money. No crisis may decrease the value of PGR except for improper maintenance. PGR must be saved in perpetuity and, ex situ should back up possible, simultaneous in-situ conservation. The collection must be built with the aim of gathering the largest genetic diversity possible. The Gene Bank collection, with the exception of conservation, must serve a broad spectrum of users. A proper determination of material is a prerequisite and any evaluation represents added value to the collection. The Prague Gene Bank wild Triticeae collection has been gathered since 1985. There are nearly 1800 accessions, belonging to 23 genera, and 133 species, of which the most important annuals represent 9 genera, 46 species and 1300 accessions. Infestation by pests and diseases was observed while collecting in wild populations. Nursery evaluation was directed mainly to powdery mildew and rusts, cereal aphids and viral infection. Infestation by aphids in field conditions was very selective towards accessions but also species and genomes. Aegilops S-genome species were the most infested by all observed aphids (Metopolophium dirhodum, Rhopalosiphum padi and Sitobion avenae). Triticum boeoticum, T. monococcum and T. timopheevii showed a high level of field resistance to viral diseases (WDV and BYDV) compared with T. urartu and T. dicoccoides. When the plants were infested by leafhopper (Psammotettix alienus) artificially, all plants were eventually diseased. The vectors showed a high level of non-preference to certain accessions. The accessions of Crithopsis delilaena and Heteranthelium piliferum showed the highest level of resistance to rusts, aphids and viral infection.
1-23.
Quantitative variation of Revolver transposon-like gene in synthetic wheat and its structural relationship with LARD element
Tomita M1, Noguchi T1, Kawahara T2
1Molecular Genetics Laboratory, Faculty of Agriculture, Tottori University, Tottori, Japan
2Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University, Muko, Japan
Revolver is a new multi-gene family dispersed like transposons in the Triticeae genomes. Revolver encompasses 3041 bp and has 20 bp of terminal-inverted repeated sequences at both ends and contains a transcriptionally active gene encoding a DNA binding-like protein. It is like Class II transposable elements. Revolver showed considerable quantitative variation through the evolution of the wheat-related species. The highest copy number of Revolver was 19,000 contained in Secale cereale (RR) and the lowest was 2,000 in hexaploid wheat Triticum aestivum (AABBDD). Next, copy numbers were determined in artificially synthetic hexaploid wheat lines between Aegilops squarrosa (DD) and tetraploid wheat species, T. dicoccoides, T. dicoccum, T. carthlicum, T. turgidum, and T. durum (AABB). Eleven lines out of 23 synthetic wheat lines showed significantly lower copies than the sum of their parental plants and 10 lines were equal to the sum, suggesting that the polyploidy was negative stress causing loss of Revolver. The members of Revolver family showed also structural variation especially in length. Revolver did not share any similarity with autonomous transposable elements. On the other hand, the LARD LTRs, which were regarded as solo LTRs of the non-autonomous retrotransposon LARD in barley, showed 60% homology to both 5' and 3' ends of some variants of Revolver. As to the region of 2 kb between the ends, LARD LTRs lacked the region of Revolver from the first exon to the middle of the first intron and resulted in non-coding sequences. LARD LTR was situated as a structural part of a LTR retrotransposon, while Revolver is a multi-gene family consisting of the exon-intron structure. Evolutionally relationship between Revolver and LARD was discussed.
1-24.
Biodiversity of the D-genome species Aegilops tauschii in Iran
Saeidi H1,2, Rahiminejad MR1, Heslop-Harrison JS2
1Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran
2Department of Biology, University of Leicester, Leicester, LE1 7RH, UK
The biodiversity and phylogeography of Ae. tauschii in Iran was assessed using morphological, molecular cytogenetic, SSR, inter-retroelement insertional polymorphism (IRAP) and AFLP markers. From the taxonomic point of view, all of the infraspecific taxa of the species were found in Iran. A high level of genetic diversity between populations was revealed by all molecular markers, but different markers revealed different level of diversity. SSR markers showed high levels of diversity without significant correlation with the taxonomic groupings. These markers are suitable for studying diversity within or between close populations. In addition to the high genetic diversity, IRAPs showed a phylogeographic pattern within the subspecies and varieties. The genetic diversity of the species significantly decreases from north to east and west of the Country, suggesting the patterns of spread and centre of diversity of the subspecies. AFLPs indicated the presence of two subgene-pools of the Ae. tauschii in Iran which was expected by other molecular markers but it was not clearly demonstrated. There was no notable difference between diversity within the subspecies (subsp. strangulata and subsp. tauschii) based on all molecular data, suggesting occurrence of high gene flow between the two subspecies in this region. In molecular cytogenetic studies, the in situ hybridization of tandem repeats (dpTa1) showed their distribution follows subspecific taxonomy. Considering the needs for introducing new characteristics and alleles for wheat improvement purposes, Ae. tauschii Iranian gene-pool is assumed to be of high importance for more investigation in the future.
1-25.
The tribe Triticeae Dumort. (Poaceae)
Yen C, Yang JL
Triticeae Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China
The tribe Triticeae is a group of taxa of Poaceae, which including very important cereal crops and very important forage grasses. Many useful and special genes are kept in the gene pool of these grasses, which are good for crop and forage improvement. So, these taxa have very high economic value. The taxonomy of Triticeae is a tool for organism recognition, an understanding of phylogenetic relationships among organisms, and also a guide for germplasm utilization. The traditional morphological taxonomy was basically based on the comparative studies of morphological characters and geographic distribution. Morphological characters are phenotypes of an organism, which are produced by functional reaction(s) between or among dominant gene(s) and environmental condition(s). External morphological characters can not reflect internal recessive inheritance. Similar environmental conditions produce convergence natural selection and different environmental conditions produced divergence natural selection. Therefore, traditional morphological taxonomy cannot avoid some mistakes in its determinations. Only cytogentic or molecular genomic analysis can avoid these mistakes. Genomic analysis can exactly determine genomic relationships and differences of species. According to recent investigations of genomic constitutions of tribe Tritceae, we recognized 30 genera in this tribe. The taxonomical changes and genomic constitution of these genera are listed in this paper.
1-26.
Synthetic wheat production for wheat breeding
Kishii M1, Mujeeb-Kazi A2
1Yokohama City University, Kihara Biological Research Institute, Maioka-cho, Totsuka-ku, Yokohama, 244- Japan
2National Institute of Biotechnology & Genetic Engineering (NIBGE), Faisalabad, Pakistan
Synthetic hexaploid wheats (SH) are a new diversity source for wheat improvement. Since 1980s, more than 1,200 have been produced by randomly crossing the wild D genome diploid donor species (Aegilops tauschii, 2n=2x=14) of various ecological origins and 51 durum wheat lines (Triticum turgidum; 2n=4x=28, AABB). This huge genetic resource captures genetic diversity of both parents and addresses a maxima of biotic and abiotic stresses. From the large number of synthetic wheats produced suitable combinations have been identified for all three rusts, spot blotch, karnal bunt, Septoria tritici, barley yellow dwarf virus, powdery mildew, drought, salinity, waterlogging, heat tolerance, improved bread making quality and some micronutrients. Few targeted synthetics were also produced by utilizing T. dicoccum and T. dicoccoides accessions. Consistent with the contributions of the ‘D genome’ exploiting the diversity of the ‘A and B genomes’ also received attention generating AAAABB and AABBBB hexaploids. The first SH production phase (upto 2004) ended by generating winter habit synthetics. Distribution of the above SH germplasm to other researchers led to the development of the ITMI (International Triticeae Mapping Initiative) population and the wheat microsatellite (SSR) map.
From a global genebank survey conducted after 2004, it turned out that 1/3 to 1/2 of the Ae. tauschii accessional holdings had been utilized for synthetic wheat production. The survey generated information on some missing geographical origin areas of Ae. tauschii. Thus in order to produce further new synthetic wheats (post 2004) and fully exploit the Ae. tauschii diversity range in a trait targeted manner, parental phenotypic and molecular characterization studies were initiated to further increase the SH number. In this later targeted approach, emmer wheat (T. dicoccum) accessions were screened for heat and drought tolerance for transfers to bread wheat. Using identified character positive accessions for these complex traits in bread wheat improvement poses the question whether genetic expression at the hexaploid level could occur. Suppression of genetic expressivity is a recognized constraint around practical utilization of SH wheats and warrants attention.
1-27.
Exploration of Triticeae resource for wheat end product quality improvement
Garg M, Tanaka H, Tsujimoto H
Laboratory of Plant Genetics and Breeding Sciences, Tottori University, Japan
Wild species of wheat are useful source of genetic variation for crop improvement. They have been utilized for improving the tolerance of wheat to different biotic and abiotic stresses. However, their potential for wheat quality has not been much investigated. In this study, we used 177 disomic addition lines belonging to 17 wild species of wheat. These lines were screened initially by poly-acrylamide gel electrophoresis for identification of addition lines carrying seed storage proteins like high-molecular-weight glutenin subunits (HMW-GSs), low-molecular-weight glutenin subunits (LMW-GSs) and gliadins from wild species. The loci of HMW-GSs, LMW-GSs and gliadins were observed on homoeologous group 1 chromosomes of wild species of wheat. Several new alleles of HMW-GSs were identified and named. Dough strength of addition lines was evaluated for 3 consecutive years and 11 addition lines with strong dough were selected. Rheological parameters of 6 addition lines revealed better quality for bread-making. Among these selected addition lines, Agropyron intermedium proteins showed best rheological characteristics followed by Ag. elongatum and Ae. searsii. Cloning and sequencing of HMW-GS genes of wild species from selected addition lines showed great diversity among these genes. Wild species whose HMW-GSs gene aligned with those of D-genome of wheat showed much better quality characteristics. Substitution lines of chromosome 1D that eventually appeared from addition lines showed very bad characteristic for bread-making quality. Preferential elimination of chromosome 1D was also observed during transfer of these alien chromosomes to selected regional cultivars and thus needs considerable attention.
1-28.
Natural variation in grain selenium concentration derived from Israeli wild barley, Hordeum spontaneum
Cheng J1, Wang F1, Yan J1,2, Xiao T3, Ning Z3, Chen G4, Nevo E2
1Institute of Triticeae Crops, Guizhou University, Guiyang, 550025, China
2Institute of Evolution, University of Haifa, Haifa, 31905, Israel
3State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang, 550002, China
4Cold and Arid Regions Environmental and Engineering Research Institute, Chnese Academy of Sciences, Lanzhou 730000, China
As the progenitor of cultivated barley, wild barley, Hordeum spontaneum is widespread in the Near East Fertile Crescent. In Israel, wild barley ranges from mesic Mediterranean areas to xeric northern and central areas of the Negev desert. Due to its rich adaptive diversity, H. spontaneum has proven to be an important genetic resource for crop improvement. In the current study, the grain selenium concentration (GSeC) originating from Israeli wild barley, Hordeum spontaneum was investigated. Ninety-four genotypes of wild barley from 9 populations were grown in the central area of Guizhou province, China. Detection of GSeC by hydride generation atomic fluorescence spectrometry (HG-AFS) method was carried out. The obtained results showed that there are remarkable differences of GSeC between and within populations. GSeC among the 94 H. spontaneum genotypes ranged from 0 to 0.387 mg.kg-1, with an average of 0.047 mg.kg-1. The highest genotype of GSeC was No. 7 originated in Sede Boqer population, while the lowest one was 25_1 originated from Atlit population. The mean value of GSeC of each population varied from 0.010 to 0.105 mg.kg-1.The coefficient of variation (CV) of each population ranged from 28% (Mt. Hermon population) to 163% (Caesarea population). Spearman’s Rho correlations between GSeC and ecogeographical data from 9 local populations in Israel were tested; and there were significant correlations between the GSeC and 12 of all 14 ecogeographic indexes. One way ANOVA indicates significant difference of GSeC among five habitat soil types. These results displayed that there are obvious genotypic differences of H. spontaneum in selenium uptake and accumulation ability. Therefore, wild barley, H. spontaneum harbors considerable differences in GSeC, and those can be used for genetic studies of barley selenium nutrition and further for crop improvement.
1-29.
Recent breeding objectives of hulled and hull-less barley for food in Japan
Yanagisawa T
National Agricultural Researcerh Center of Western Region (WeNarc), Barley Research Team, Senyu 1-3-1, Zentsuji, Kagawa 765-8508, Japan
In Japan, barley is used not only for alcohol beverages but food such as miso, rolled barley and barley tea. Breeding efforts for genetic improvement for seed composition and grain quality, as well as increasing yield and introducing disease resistance, is important breeding objectives of barley for food in Japan.
Last year, two-rowed hull-less barley “Yumesaki-boshi” was released. This cultivar is resistant to barley yellow mosaic virus and powdery mildew, and is moderately resistant to scab. Since grain size is large, pearled grain is high whiteness; this cultivar is tentatively used for rolled barley.
Barley is susceptible to a browning reaction after heating and browning reaction is correlated to its polyphenol content. Proanthocyanidin is a kind of polyphenol, so proanthocyanidin-free mutants (ant 28) were useful to reduce the browning reaction in boiled pearled barley. “Tochinoibuki” and “Shirotae-Nijo” (two-rowed hulled) were released last year. These cultivars will be used for retort-packed food.
(1,3) (1,4)-beta-D-glucan (beta-glucan) is major components of polysaccharides in cell walls of barley endosperm. Beta-glucan is dietary fiber, is favorable for human foods because it lowers cholesterol. High beta-glucan contents (>10%) hull-less barley is recently tested in the performance test for recommended variety.
Recently whole barley grain barley and certain dry milled barley grain products are appropriate sources of beta-glucan soluble fiber for the health claim in USA, so barley has the possibility of the functional foods.
1-30.
Waterlogging and salinity tolerance in wild Hordeum species: physiological basis and prospects for use in wheat improvement
Colmer TD1,2, Islam AKMR1,3
1Future Farm Industries CRC, The University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia
2School of plant Biology, The University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia
3School of Agriculture and Wine, The University of Adelaide, Waite Campus, Glen Osmond, SA, 5064, Australia
The genus Hordeum contains many wild species that grow in diverse habitats (non-saline to saline, dryland to wetland). We evaluated waterlogging and salinity tolerances, and selected physiological traits, in a range of Hordeum taxa. Species (and sub-species) with the X genome and several with the H genome were waterlogging tolerant, whereas those with the I genome, Y genome and the remaining H genome species, were much less tolerant. Waterlogging tolerant species displayed traits for improved internal aeration of roots. H. marinum (X genome) was not only one of the most waterlogging tolerant species, but was also salt tolerant. H. marinum maintains low leaf Na+ and Cl- concentrations, even when exposed to levels of salinity that can kill wheat. H. marinum-wheat amphiploids have been produced, firstly with Chinese Spring and recently with several Australian varieties. The amphiploids show improved salinity and waterlogging tolerances, with better root aeration and regulation of leaf Na+ and Cl- concentrations. The current amphiploids are in H. marinum cytoplasm, and suffer some cytoplasmic male sterility. Selected amphiploids will be transferred to wheat cytoplasm to restore normal fertility. The aim initially is to develop feed wheat for moderately affected salt land. Furthermore, in a parallel program, 6 out of the 7 possible disomic additions lines (1Hm, 2Hm, 4Hm, 5Hm, 6Hm and 7Hm) of individual H. marinum chromosomes to Chinese Spring wheat have been produced. These addition lines will be useful in determining the chromosomal controls of salt- and waterlogging-tolerance in H. marinum, as expressed in a wheat background.
1-31.
Biological Nitrification Inhibition (BNI) potential in Triticeae
Subbarao GV, Ishikawa T, Ito O
JIRCAS (Japan International Research Center for Agricultural Sciences), Ibaraki 305-8686, Japan
Nitrification is the key process in the global nitrogen cycle that results in the formation of nitrate through microbial activity, negatively affecting the availability of nitrogen to plants, causing low nitrogen-use efficiency in agricultural systems. The ability of certain plants to suppress soil nitrifier activity by releasing inhibitors from roots, a plant function, termed “biological nitrification inhibition (BNI)”. Using recombinant luminescent Nitrosomonas europaea to quantify BNI release, we found that wheat wild relative Leymus racemosus releases about 20 times more BNIs than cultivated wheat and effectively suppresses NO3- formation in soil. The high BNI-capacity in L. racemosus is controlled by chromosome Lr+n, which was introduced into Leymus-wheat chromosome addition lines via inter-specific crosses. The high BNI release capacity from Leymus was successfully expressed in wheat lines. Our recent results from barley indicate substantial genetic variability for BNI-capacity. Unlike synthetic nitrification inhibitors that block only the AMO pathway, BNIs block both AMO and HAO enzymatic pathways of Nitrosomonas, making the inhibitory effect more stable. Recent field evaluations of high-BNI capacity tropical pasture grasses (Brachiaria spp.) demonstrated substantial reductions (>90%) in soil nitrification rates and nitrous oxide emissions. Given the wide-spectrum of genomic structure, the unique capacity for inter-generic gene flow and the ability to form allopolyploid genomes, Triticeae could offer a range of genetic tools/options to exploit and introduce sufficient BNI-capacity into wheat, barley and rye. Potential implications from such an approach in facilitating a shift towards sustainable NH4+-dominated cereal production systems is the subject of discussion.
1-32.
Reduced transcript levels at the Bot3 locus in barley (Hordeum vulgare L.) confer increased tolerance to high boron supply
Schnurbusch T1,3, Hayes J1, Tyerman SD2, Baumann U1, Pallotta M1, Ramesh S2, Langridge P1, Sutton T1
1Australian Centre for Plant Functional Genomics, The University of Adelaide, Waite Campus, PMB 1 Glen Osmond, SA 5064, Australia
2The University of Adelaide, School of Agriculture, Food & Wine, Australian Centre for Plant Functional Genomics, Waite Campus, Urrbrae, SA 5064, Australia
3Present address: Leibniz-Institute of Plant Genetics and Crop Plant Research (IPK), Genebank Department, Corrensstr. 3, D-06466 Gatersleben, Germany
Barley (Hordeum vulgare L.) is an annual cereal grain and has been ranked number four among the staple foods of the world. Today’s cultivated barleys are not well adapted to high soil boron (B) although toxicity to B has been known for some time. The Algerian landrace Sahara 3771 proved of being highly tolerant to B and thus, represents one of the most B-tolerant barleys currently known. It carries four quantitative trait loci (QTL) conferring tolerance to toxic B conditions. One gene (Bot1) underlying the tolerance QTL on chromosome 4H of barley has recently been identified and is a putative membrane-bound B transporter with similarity to bicarbonate transporters in animals; it functions as an efflux transporter to move B out of the plant (Sutton et al. 2007; Science 318:1446 ff.). Bot1 was the first B tolerance gene to be identified in plants. In this work, we describe the cloning of a second high B tolerance QTL in barley, mapping to the 6H B tolerance locus, Bot3. Using heterologous expression systems in yeast and Xenopus oocytes, we show here that this gene can facilitate the transport of B across membranes. Higher tolerance to B in Sahara 3771 is mediated through lower transcript levels of Bot3 in root tips of barley plants possibly owing to a repeat insertion into the promoter region of Bot3 approximately 2 kb upstream of the start codon. Moreover, we observed lower shoot B accumulation in a rice (Oryza sativa L.) mutant possessing a point mutation in the orthologous rice transporter gene. Based upon our results we conclude that under high soil B Bot3 entails lower shoot B accumulation and thus, effectually aids to higher B tolerance in Sahara 3771.
1-33.
Genetic targeting of drought sensitive gene eibi1 of wild barley (Hordeum spontaneum)
Chen G1,2, Pourkheirandish M1, Sameri M1, Wang N1, Duan Z2, Shi Y2, Li2, Nevo E3, Komatsuda T1
1Plant Genome Research Unit, National Institute of Agrobiological Sciences, Tsukuba 305-8603, Japan
2Ecology and Agriculture Department, Cold and Arid Regions Environmental and Engineering Institute, Chinese Academy of Sciences, Lanzhou 730000, China
3Institute of Evolution, University of Haifa, Mount Carmel, Haifa 32905, Israel
The plant cuticle plays multiple roles in the protection of plant from various environmental stresses, especially the drought stress. The spontaneous cuticle mutant eibi1 derived from Israeli wild barley was characterized and fine mapped in the present study. eibi1 showed the highest relative water loss rate among the known wilty mutants, which indicates that eibi1 is one of the most drought-sensitive mutants. eibi1 was neither an ABA-deficient nor an ABA-insensitive mutant. The eibi1 leaves had a larger chlorophyll efflux rate in 80% ethanol than the wild-type leaves. The stomatal movement of eibi1 was normal. eibi1 had a more than 10 times larger transpiration rate than the wild type in the dark. These lines of evidences indicated that eibi1 was defective in the cuticle. The scanning electronic microscope (SEM) analysis revealed that the cuticular layer of eibi1 leaf was broken, not continuously covering the leaf surface, which caused endless water loss. This SEM result confirmed the conclusion that eibi1 was defective in the cuticle. The eibi1 caused pleiotropic effects including semi-dwarf, low fertility, kinked peduncle, drought sensitive, and hulless seed. eibi1, a monogenic and recessive mutant, was mapped to the pericentromeric region of chromosome 3H. To facilitate map-based cloning of EIBI1, we conducted a high resolution mapping of eibi1 with 1682 F2 individuals of Morex x eibi1. Barley-rice synteny was employed to search for ESTs for eibi1 mapping and to identify candidate genes for eibi1. Comparison of the barley high resolution genetic map and rice physical map revealed the inversion of the eibi1 region in barley against its corresponding region in rice. However, the collinearity within this inverted region was well conserved. eibi1 was located in a region of 0.11 cM in barley genetic map which corresponding region in rice chromosome 1 physical map was 112.8 kb.
1-34.
Genetic engineering in cereals: Current technologies for the elucidation of gene functions
Kumlehn J
Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, Germany
The development of reliable Agrobacterium-mediated transformation technologies for Triticeae cereals has greatly stimulated a variety of approaches to the functional analysis of genes and the generation of genetically engineered breeding lines. In the Triticeae model species barley, three types of regenerable recipient cells or tissues have been successfully employed for the stable Agrobacterium-based DNA-transfer, each having specific advantages. While the use of immature zygotic embryos results in exceedingly high efficiency of transgenic plant formation, embryogenic pollen cultures enable us to instantly produce true-breeding transgenic lines, whereas isolated ovules facilitate the generation of transgenic plants without use of a selectable marker. In addition to the barley platform established, Agrobacterium-mediated transformation is also perfomed in winter and spring wheat as well as in rye. To make the creation of plasmids for the transformation of cereals more efficient, a set of generic binary vectors for over-expression and RNAi-approaches has been generated. The vector set permits a convenient integration of any gene or gene fragment by GATEWAY-based recombination and comprises derivatives with various promoters. As a result, constitutive as well as endosperm-, epidermis- and egg cell-specific expression systems have been established. Further promoters and selectable markers of choice can be readily integrated due to the vectors' modular configuration. Through coupling genetic transformation with haploid technology, we have been able to rapidly produce homozygous transgenic barley lines. Based upon the enabling technology established, we have embarked on numerous projects aiming to functionally analyse candidate genes, and to generate transgenic barley lines with improved performance.
1-35.
Map based cloning of dormancy QTL in barley
Sato K1, Matsumoto T2, Ooe N1, Takeda K1
1Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan
2National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba 305-8602, Japan
Seed dormancy is a trait of wild barley to escape drought in the summer of arid condition. Dormancy in cultivated barley has different crop functions e.g. delays malting process or prevents pre-harvest sprouting. Thus, the cloning of dormancy gene in barley contributes to understand domestication process and optimize the trait for efficient uses. Rates of seed germination were used to evaluate dormancy on physiologically matured grain samples after dried and stored frozen until use. Many genetic factors controlling seed dormancy has been reported as quantitative trait loci. Among these loci, a locus on the centrometic region of chromosome 5H (Qsd1) has been most frequently identified and showed largest effect among mapping populations. This QTL was identified on the EST map of Haruna Nijo (H. vulgare ssp. vulgare) and wild barley H602 (H. vulgare ssp. spontaneum) by both doubled haploid population and recombinant chromosome substitution lines (RCSLs). At least four QTLs are segregating in this cross. RCSLs having only Qsd1 in the segment of wild barley on the genetic background of Haruna Nijo were selected and a total of 919 B3F2 plant was used to develop a high resolution map of the QTL. By using rice genome and barley cDNA sequences, possible barley transcript based markers were generated to map the locus. By selecting heterozygous plants of B3F4, 4,792 B3F3 plants were genotyped to evaluate recombinants between QTL and transcript based markers. After sequencing BAC clones of Haruna Nijo and H602, the possible mutations were identified between two gene sequences. Functional analyses of these genes are underway.
1-36.
High frequency of karyotype variation revealed by sequential FISH and GISH in breeding population of plateau perennial grass forage Elymus nutans
Dou QW1, Chen ZG1, Liu YA1, Tsujimoto H2
1Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Plateau Institute of Biology, the Chinese Academy of Sciences, Xining, 810008, China
2Faculty of Agriculture, Tottori University, Tottori, 680-8550, Japan
Karyotyping was conducted to 12 random selected samples of grass forage Elymus nutans (StHY, 2n=42) which are widely planted in QingHai-Tibet plateau in China by sequential FISH and GISH. The results showed that genome constitutions of 12 samples were different from each other and nearly each chromosome included high frequent variants. The St genome shared the highest number of variants of 42, 38.2% of total genome variants; while the Y genome shared the lowest number of variants of 33, 30.0% of total. High frequency of translocation was revealed that 7 of 12 samples, 58.3% of total, carried non-Robertsonia translocation chromosomes between different genomes. 4 types of different translocations were demonstrated. 4 samples carried the type 1 and 3 samples carried each of the other 3 types respectively. High frequency of heterogeneous karyotype was revealed that 7 samples, 58.3% of the total, showed that one or more than one chromosome lacked the homologous chromosomes which shared the same or similar repetitive sequence FISH patterns.
1-37.
Revealing genetic diversity in closely related A-genome diploid wheat species (Triticum boeoticum, T. monococcum, T. urartu) by retrotransposon display
Konovalov FA1, Goryunova SV1, Shaturova AS1, Fisenko AV1, Melnikova NV1, Kudryavtsev AM1, Goncharov NP2
1Vavilov Institute of General Genetics, GSP-1, Gubkina 3, 119991 Moscow, Russia
2Institute of Cytology and Genetics, Lavrentiev ave. 10, 630090 Novosibirsk, Russia
LTR retrotransposons constitute a rapidly evolving part of plant genomes due to their transpositional activity and susceptibility to deletions. Retrotransposon-based DNA markers are known to provide a high level of polymorphism which can be used for discriminating closely related accessions. If a particular retrotransposon family does not have a strong target site preference, it can be assumed that an ancestral state (absence at a given site) is known for each inserted copy and that independent insertion events into the same site are unlikely to occur.
We have explored the usability of retrotransposon display (also known as SSAP method, Sequence-Specific Amplification Polymorphism; Waugh et al., 1997) to infer the relationships within a group of A-genome diploid wheat species Triticum boeoticum, T. urartu, T. monococcum and T. sinskajae. The presence/absence data for 200 polymorphic SSAP bands representing the insertion sites of Sasanda and BARE-1/WIS retrotransposon families were obtained for 49 accessions. For a given accession set the majority (>90%) of bands that separated in a sequencing gel were polymorphic. The marker system was also successfully tested on several representatives of Triticeae and closely related taxa, including wheat species of different ploidy, Aegilops, Hordeum, Secale and Avena species. Our modified SSAP procedure is relatively tolerant to DNA amount, and the bands can be visualized by silver staining. Various approaches to retrotransposon display data analysis have been compared, including Dollo parsimony and several distance-based methods. Diploid wheat phylogeny issues are discussed.
1-38.
Revealing seed storage metabolism from genetical genomic and systems biology approaches
Sreenivasulu N, Pietsch C, Radchuk V, Röder M, Weschke W, Wobus U
Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK), Corrensstraβe 3, D-06466 Gatersleben, Germany
In our ongoing efforts we developed large scale EST resources (Zhang et al., 2004) and throughput gene expression profiling platforms in barley (Sreenivasulu et al., 2006; Sreenivasulu et al., 2004; Sreenivasulu et al., 2002), which have brought substantial progress in elucidating biochemical pathways of barley seed metabolism in barley (Sreenivasulu et al., 2008a; 2008b, Neuberger et al., 2008, Strickert et al., 2007). Very recent findings shed light on the interplay of many cellular and metabolic events that are coordinated by a complex regulatory network during barley seed development. Studying expression data of nearly 12,000 seed-expressed genes revealed, for instance, the participation of tissue-specific signaling networks controlling ABA-mediated starch accumulation (via SNF1 kinase and a set of transcription factors) in the endosperm and participation of ABA-responsive genes in establishing embryo desiccation tolerance. In an attempt to gain insight into the underlying genetic factors that govern differences in storage product accumulation we compared changes in gene expression patterns during seed development among 30 lines carrying defined wild barley introgressions and dissected the candidate regulatory genes involved in altering the process of storage product accumulation through genetical genomics approach. With the focus of using the ‘developing seed’ as model for systems biology studies we further investigated transcriptional and metabolic networks during grain development, developed models of the developing barley grain and implemented magnetic resonance-based techniques. These tools will be used to explore transgenic model systems for identifying key regulatory networks related to seed quality traits.
1-39.
Hardness locus sequence variation in association with grain quality in spring barley (Hordeum vulgare L.)
Turuspekov Y1,4, Beecher B2, Darlington Y1, Bowman J3, Blake TK1, Giroux MJ1
1Dept. of Plant Sciences and Plant Pathology, 119 Plant BioSciences, Montana State University, Bozeman, MT 59717-3150, USA
2E.202, FSHN Facility East, USDA-ARS, Washington State University, Pullman, WA 99163, USA
3Depty. of Animal and Range Sciences, 230C Linfield Hall, Montana State University, Bozeman, MT 59717, USA
4current address: Inst. ofPlant Biology and Biotechnology, NCB RK, Almaty 050040, Kazakhstan
Grain hardness has an important impact on the end-use quality of cereal grains. The Hardness (Ha) locus in barley contains the Hina, Hinb-1, Hinb-2, and Gsp genes and was shown to be associated with grain hardness and dry matter digestibility (DMD) variation. In this work, 73 spring barleys accessions selected for DMD variation were assessed for variation in DMD, seed quality, and Hardness (Ha) locus component gene alleles. In order to determine whether Ha is associated with grain quality traits, we assessed the relationship of the Ha locus in the presence or absence of head type (2 or 6-row) variation. To accomplish this, the barley Ha locus component genes (Hina, Hinb-1, Hinb-2, and Gsp) were sequenced from each accession and sorted by prevalence. The most common Ha haplotype (HINA, HINB-1, HINB-2 alleles) was present in 42 accessions with the remaining 39 dispersed over 24 haplotypes. Seeds from two rowed accessions with the most common Ha haplotype were significantly softer in grain texture (P<0.001) and had increased starch content (P<0.001) and DMD (P<0.05). The results indicate that selection for individual Ha locus haplotypes may be useful in modifying seed size, DMD, and starch content in 2 rowed barley.
1-40.
Mutual interchange of genetic variation and genomic information between wild and cultivated species in Triticeae
Tsujimoto H
Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan
The increase of wheat yield by breeding has slowed down in the last decade. It will not be easy to solve the food crisis which we are facing today without another green revolution. Wild species of the tribe Triticeae grow in a wide range of environments around the world and have large genetic variation. This variation should have been efficiently used for wheat improvement. But the method to introduce wild chromosome segments to wheat chromosomes has got behind without using recent genomic information. New molecular breeding system needs to be developed to introduce only small chromosome regions to wheat using accumulated information of wheat and barley genomes and that of meiosis studied in model organisms. The problem that occurs in transferring genes to different genomes is the co-transfer of undesired deleterious genes located near the aimed gene. Application of fine molecular maps of wheat and barley will make it possible to pin point the location of the desired gene and to transfer only a smallest portion to a wheat chromosome. Homoeologous pairing may be induced by ph1b or PhI genes, but crossing and chromosome observation is necessary. Isn’t it possible to induce homoeologous pairing by chemical and physical stimulation? In this presentation, I will discuss the problems and possible solutions to these problems occurring when the genetic variation of wild Triticeae species is used for wheat improvement.
1-41.
Development and cytogenetic analysis of Hordeum chilense chromosome 4 introgression lines into durum wheat
Prieto P, Ramírez MC, Martín A
Consejo Superior de Investigaciones Científicas (CSIC), Apartado 4084, 14080 Córdoba, Spain
Wheat is one of the most important food crops in the world and the wild specie Hordeum chilense Roem. et Schult is a valuable genetic resource for wheat breeding. In fact H. chilense carries interesting agronomic genes like resistance to the root-knot nematode Meloidogyne naasi on chromosome 1HchS; tolerance to salt on chromosomes 1Hch, 4Hch and 5Hch; resistance to Septoria on chromosome 4Hch; and high carotenoid pigment content and resistance to common bunt, both located on chromosome 7Hch. In breeding programs addition lines are useful as starting point for the transfer of agronomic traits to wheat, but the introgression of specific characters requires the identification of recombinants that remove unwanted regions of the H. chilense chromosome. With the aim of introgress resistance to Septoria in durum wheat we have developed inter-specific crosses between a 4Hch(4B) substitution line in bread wheat (T. aestivum, 2n = 6x = 42, AABBDD) and the 4D(4B) substitution line in durum wheat cv. Langdon (T. turgidum, 2n = 4x = 28, AABB). Backcrosses by durum wheat cv. Yavaros, which is a better adapted cultivar to our agro-climatic region, have been also developed. A set of monosomic and disomic 4Hch(4B) chromosome substitutions and additions have been generated in durum wheat. Alien chromosome segments have been reduced in size by further spontaneous translocations between chromosome 4A wheat and chromosome 4Hch of H. chilense. We present the identification and characterisation of all these introgression lines in durum wheat by multicolour fluorescent in situ hybridisation.
1-42.
Transcriptional landscape of malting barley, Haruna Nijo - using a custom-made oligoarray from cDNA sequences
Matsumoto T1, Ikawa H2, Fujii N1, Tanaka T1, Sakai H1, Itoh T1, Nakamura S3, Sato K4
1National Institute of Agrobiological Sciences (NIAS), Kan-non-dai 2-1-2, Tsukuba, Ibaraki 305 8602, Japan
2STAFF-Institute, Ippaizuka 446-1, Kamiyokoba, Tsukuba, Ibaraki 305-0854, Japan
3National Institute of Crop Sciences (NICS), Research Team for Barley and Wheat Biotechnology, Kan-non-dai 2-1-18, Tsukuba, Ibaraki 305 8518, Japan
4Research Institute for Bioresources Okayama University, Chuo 2-20-1, Kurashiki, Okayama 710-0046, Japan
In order to understand barley transcriptome under the normal and the stressed conditions, we have designed 60-nucleotide oligomers from 3’ end sequences from the ca. 36 000 full-length cDNA clones of two-rowed malting barley, Haruna Nijo, and these were used for 36K Agilent-type oligoarray construction. Microarray experiments indicated that about 1/8 of the genes tested were drastically influenced under either of the 42 stress conditions. Using clustering function of the GeneSpring analysis software, these affected genes are classified into several categories, such as co-expressed genes in root with salt (NaCl), dehydration, and ABA stresses (such as ABA-inducible protein PHV A1 and ACC oxidase), indicating there are common responsive pathway to these diverse stimuli. While we found known annotation in the stress responsive genes, many other genes had no known functions in barley, rice, and Arabidopsis. Functional information revealed in this study will not only endow novel understanding to barley gene function, but also give some hints to elucidate biological functions of the orthologous genes in grasses. This work is supported by the grant from the Ministry of Agriculture, Forestry and Fisheries, Japan (MAFF: TRC1008).
1-43.
Salinity tolerance and sodium exclusion in genus Triticum
Shavrukov Y, Langridge P, Tester M
Australian Centre for Plant Functional Genomics, University of Adelaide, Urrbrae, SA 5064, Australia
The ability of plants to exclude sodium from the shoot is one of the major components of salinity tolerance. The mechanism of sodium exclusion is presented in genus Triticum and the considerable variability in sodium exclusion within different species is demonstrated. The diploid species T. monococcum revealed a large (50-fold) variability in sodium exclusion in contrast to T. urartu, which was significantly less variable (10-fold). These species with the A genome are known to be salt sensitive, whilst T. (Aegilops) tauschii, a diploid species with the D genome, was very salt tolerant, but had only moderate variability in sodium exclusion (10-fold). The tetraploid species T. turgidum ssp. durum (both cultivated and landraces) and wild emmer T. dicoccoides (all with the AB genome) showed a range of variability in both salinity tolerance and sodium exclusion. The general pattern (from most sensitive and with highest Na+ accumulation) was as follows: durum (cultivated) < durum (landraces) < wild emmer. Cultivated durum wheats had minimal or no variability, whereas landraces of durum wheats had greater variability, with two excellent genotypes having been identified which combine very low sodium accumulation with very high salinity tolerance. Wild emmer was extremely variable. Hexaploid bread wheat, T. aestivum with the ABD genome, is known to be more salt tolerant, having an effective mechanism for sodium exclusion but only low variability. As a result of spontaneous hybridisation during their origin, cultivated durum and bread wheats have a limited gene pool and low variability in salinity tolerance. Introgression of new genes from different progenitors and relatives of Triticum that cope better with salt stress can significantly improve salinity tolerance in both cultivated durum and bread wheats.
1-44.
Submergence tolerance in Hordeum marinum
Malik AI1,2,3, Pedersen O1,4, Colmer TD1,2
1School of Plant Biology, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia
2Future Farm Industries CRC, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia
3Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
4Freshwater Biological Laboratory, University of Copenhagen, Helsingørsgade 51, 3400 Hillerød, Denmark
In the tribe Triticeae, the genus Hordeum contains many wild species that grow in a range of dryland to wetland habitats, and those growing in wet areas could naturally experience flash flooding. Submergence tolerance in three H. marinum accessions (H21, H90 and H546) was tested for 7 days during the vegetative stage. Waterlogging alone (i.e. O2-deficient root-zone) had no effect on whole plant growth of H21 and H90, however, H546 was reduced to 82% of the aerated control. Complete submergence of the shoots ([CO2] of 200 μM in floodwater) reduced whole plant growth to 61%, 31% and 50% of the aerated controls of H21, H90 and H546, respectively. Underwater net photosynthesis by submerged leaves was about one-third of that in air. Tissue sugars were reduced in submerged plants, but not in waterlogged plants. Porosity (% gas volume per unit tissue volume) in leaf sheaths and roots enabled diffusive O2 transport from floodwaters during darkness, and of O2 produced during photosynthesis during light periods, to the roots in a hypoxic root medium. The adverse effects of submergence were reduced when dissolved CO2 in the floodwater was increased. In summary, the study shows that some accessions of H. marinum are highly tolerant of waterlogging (i.e. shoots in air, roots in O2-deficient medium), and there is also diversity in tolerance to complete submergence.
1-45.
USDA-ARS gene, genomic, and trait discovery research in perennial Triticeae grasses
Larson SR, Bushman BS, Mott IW, Wang RR-C, Jensen KB, Robins J, Jones TA
USDA-ARS Forage and Range Research Laboratory, Utah State University, Logan, UT, 84322-6300, USA
Long-term grass breeding efforts at the USDA-ARS Forage and Range Research Laboratory, traditionally focused on perennial Triticeae range grasses, have been expanded to include the development of molecular markers and genetic maps that can be used to investigate the genetic control of functionally important perennial grass traits. Genetic maps constructed from tetraploid interspecific hybrids of tall caespitose Leymus cinereus and relatively short rhizomatous Leymus triticoides have been used to identify chromosome regions associated with plant height, growth habit, flowering, seasonal biomass accumulation, seed shattering, seed germination, biotic and abiotic stress resistance, inflorescence traits related to seed production, and forage quality (fiber, protein, and mineral content). A total of 27,273 expressed gene sequence tag (EST) unigenes have been isolated and sequenced from Pseudoroegneria (8780), Elymus (7212), and Leymus (11281). Moreover, 1375 Pseudoroegneria, 442 Elymus, and 1798 Leymus EST simple sequence repeat (SSR) have been identified. PCR primers flanking these EST-SSRs have been tested for amplification and polymorphism on parental genotypes of existing or proposed new mapping populations. A total of 375 Leymus EST-SSR markers amplified one or more segregating markers in the Leymus mapping populations. These Leymus EST markers were used to align the Leymus linkage groups to the rice genome sequence and identify homoeologous groups relative to wheat, barley, and other Triticeae genomes. Bacterial artificial chromosome (BAC) genomic DNA libraries representing approximately 6.1 haploid genome equivalents of tetraploid Leymus have also developed and used to compare homoeologous genomic DNA sequences containing the rice lax and maize barrenstalk1 orthogene. Likewise, genetic maps are being constructed from interspecific hybrids of caespitose Elymus lanceolatus and rhizomatous Elymus wawawaiensis. Other traits of interest in the Elymus mapping populations include resistance to billbug (Parvulus parvula) feeding.
1-46.
Plasmon analysis in the Triticum-Aegilops complex
Tsunewaki K
6-14-10 Kasagadai, Nishi-ku, Kobe, Hyogo 651-2276, Japan
This paper reviews past results of our group on plasmon analysis of the Triticum-Aegilops complex. Planned content of the talk is as follows: (1) Classification and characterization of the plasmon; genetic effects to 47 plasmons on 21 wheat characters were analyzed using 563 eu- and alloplasmic lines, in which 12 common wheat genotypes and 47 Triticum-Aegilops plasmons were combined in all possible combinations, except one lethal combination, and characteristics of the individual plasmons were clarified, based on which they were classified into 16 groups. (2) Classification of the plastomes and chondriomes; their genetic diversities were studied by RFLP analyses of chloroplast and mitochondrial DNAs, respectively. Based on the data, genetic distances between all pairs of plasmons were estimated for both the plastomes and chondriomes, based on which their phylogenetic trees were constructed. Sites of some plasmagene mutations were speculated in the chondriome phylogeny. (3) Genetic diversity of the plasmon; combining the results of the above works, 48 plasmons were classified into 19 types, to which plasmon symbols were designated. Characteristics of individual plasmon types will be described. Differentiation of the plasmon at the diploid level and maternal lineages of polyploid species became evident. Origin of some polyploid species were assumed to be more recent than some other relatives, three examples being Timopheevi vs. Emmer in the tetraploid wheat, Ae. ventricosa vs. Ae. crassa of the Vertebrata, and Ae. triaristata vs. Ae. ovata of the Polyeides section, of the genus Aegilops. Referring to the genome symbols designated by previous workers, the genome-plasmon constitutions of all Triticum-Aegilops species became elucidated, owing to which species relationships of this complex became clear in most part for both genome and plasmon, the first case at generic level throughout the plant and animal kingdoms. At the end, some plasmon-related problems remained unsolved will be pointed out.
1-47.
The barley coordinated agricultural project (CAP): integrating genomics with breeding
Muehlbauer GJ
Barley CAP consortium and AGOUEB consortium
The barley coordinated agricultural project (CAP) is an integrated project that leverages community strength in genomics, statistics, computer science and breeding to enhance the efficiency of barley breeding. The basic idea is to genotype and phenotype advanced breeding lines from 10 U.S. barley breeding programs and to utilize the combined datasets to conduct association-based analysis to identify quantitative trait loci (QTL). During the first phase of the project we worked with our international collaborators and mapped 2,943 single nucleotide polymorphisms (SNP). From these mapped SNPs, an international SNP genotyping platform was developed. To date, 2880 breeding lines have been genotyped with over 3,000 SNPs and phenotyped for over 40 traits including agronomic, disease resistance, and food and malting quality. The SNP genotyping has provided the opportunity to understand linkage disequilibrium (LD) in U.S. barley breeding germplasm, and to understand the genetic relationships between breeding programs. The model that fits the 10 U.S. barley breeding programs is seven populations with LD ranging from 20-30 cM. All data have or will be deposited in a centralized database called The Hordeum Toolbox. We used the combined genetic and phenotypic datasets to detect QTL for a variety of traits including malting quality, winterhardiness, dormancy, preharvest sprouting, drought tolerance and resistance to Fusarium head blight, spot blotch, net blotch, African stem rust (Ug99), and common root rot. The success of identifying marker-trait associations has lead to the first stages of implementing marker-assisted selection. An example of the breeding utility of mapping resistance to Fusarium head blight will be described.
1-48.
Features of East Asian barley and their genetic analyses
Sato K, Takeda K
Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan
Cultivated barley was considered domesticated in Middle East and distributed to East Asia. Abundant morphological variation, which was exotic to Occidental barley germplasm, has been found in East Asian barley landraces. The cause of this morphological variation is still not understood. However, the uses of barley flour in Tibetan people and steamed pearled grain in East Asian area might keep these variation unselected. The growing condition of East Asian barley is also very different from the one in Occidental barleys. An obvious different growing condition in East Asian barley is the cultivation in the paddy field after rice. The habit is still common in Japan, Korea and southern part of China. These areas have rainy season in early summer caused by the Asian monsoon climate. There are some typical abiotic and biotic stresses due to these special growing conditions. Since barley cultivars are grown in the paddy field where soil moisture is constantly high, plants are continuously exposed to excessive water. There might be repeated severe selections in this condition for barley and we have some promising tolerant accessions from East Asia. There is a special semi-drawf barley genotype uzu which has been mainly distributed in western part of Japan. This group of barley show tolerance to lodging in wet field condition. Higher precipitation after flowering also promotes fungal disease development in barley. Fusarium head scab attacks barley spikes and damages grains by mycotoxin contamination. The promising source of resistance to this disease has been also found in East Asian barley germplasm. Some results of genetic analysis of these stress tolerances will be presented.
2. Poster Presentation
2-1.
DNA markers: another tool in the toolbox
Bushman BS1, Barkworth ME2
1USDA-ARS Forage and Range Research Lab, Logan, UT, USA
2Intermountain Herbarium, Utah State University, Logan, UT, USA
A scientific name has several meanings, and is often taken for granted by the larger scientific community. However, if ambiguity exists for taxa, the scientific names may be incorrect and lead to erroneous conclusions with lasting impact. Nowhere is this a greater problem than within the grasses, and Triticeae is not exempt. In this presentation we discuss instances of taxonomic confusion, situations that give rise to taxonomic confusion, and how the use of molecular markers can add another valuable tool to the toolbox of a taxonomist. We highlight situations where one or few genes are responsible for traits with tremendous phenotypic differences, where lasting hybrid populations give rise to new species names, where overlapping characteristics preclude diagnostic morphological characters but can be resolved with DNA sequences, and where DNA sequences and morphology suggest different taxonomic interpretations. Our conclusion is that a molecular biologist working with grasses that have taxonomic ambiguity should have a plant taxonomist as a friend, and vice-versa.
2-2.
Phylogenetic and phylogeographic analyses of Hordeum murinum (Poaceae)
Jakob SS, Blattner FR
Leibniz Institute of Plant Genetics and Crop Research (IPK), D-06466 Gatersleben, Germany
The Hordeum murinum taxon complex consists of diploid H. murinum subsp. glaucum and the two polyploid subspecies murinum (4x) and leporinum (4x, 6x). The taxa are native in the Mediterranean and adjacent areas of Eurasia and northern Africa, and were introduced worldwide as weeds in man-made habitats. For the polyploids it is unclear if they originated via (segmental) allopolyploidization or are autopolyploids.
AFLP analysis of individuals covering all taxa, ploidy levels, and the entire native distribution resulted in two clades, separating diploid subsp. glaucum from all polyploids. Also extensive sequencing of cloned PCR products of the nrDNA ITS region showed a clear separation of diploid from polyploid cytotypes with only few diploid alleles found within single polyploid individuals. As this could be a result of recent hybridization, we sequenced also a single-copy nuclear gene (TOP6). This locus clearly showed that subsp. glaucum contributed to the polyploids. Phylogenetic analysis revealed that the second parent belongs to the H. murinum genome group (Xu genome) but is nowadays extinct. This latter species was most probably the maternal parent of the polyploids, as their chloroplast type is clearly different from the types occurring in extant diploid individuals. None of the markers used was able to separate subsp. murinum from leporinum although these taxa are morphologically and geographically clearly separated, which might indicate a young age of these taxa and cytotypes.
2-3.
Levels of study of StH-genomic Elymus species of Asian Russia and North-Eastern Kazakhstan in connection with a problem of “species-phantoms”
Gerus DE, Agafonov AV
Central Siberian Botanical Garden, SB RAS, Zolotodolinskaya st., 101, Novosibirsk, 630090, Russia
There are 72 species of the genus Elymus L. in the former Soviet Union according to S. Czerepanov (1995).It is difficult to determine an exact number of species for the vast area of due to contradictory data of the authors. North-Eastern Kazakhstan is the area closely related in respect of floristics with the Altai-Sayan region of Russia. Fourteen new Elymus species were described from North-Eastern Kazakhstan over the last 17 years. Species from Asian Russia and North-Eastern Kazakhstan can be characterized by the following levels of study: a) species well studied by the methods of biosystematics with data on the genetic analyses of some morphological characters; b) species studied by means of genetic markers (DNA, grain proteins, histone H1); c) species widely represented by herbaria; d) “species-phantoms” not studied, known only by type-specimens, without mature seeds. The greatest number of “species-phantoms” was described from North-Eastern Kazakhstan. Only 44 species of the genus Elymus have StH constitution or unknown one. Among them, 20 species belong to level a, 22 species – b, 22 species – c, 18 species – d. Taxonomic problems connected with study of “species-phantoms” will be discussed in the poster.
2-4.
Taxonomy and inter- specific relationships of Agropyron Grant. in Iran
Hasheminejad N1, Saeidi H1, Yoosofi M2, Rahiminejad MR1
1Dept. of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran
2Dept. of Biology, University of Payame Noor University, Najafabad, Isfahan
The genus Agropyron has been restricted to: A. cristatum (L.) Gaertn. and A. desertrum (Fisch.) Schultes with seven infra-specific taxa in the former in Iran. Taxonomic status and inter taxa relationships of the genus in Iran were examined using taxonomic, cytotaxonomic and molecular (SSR markers) studies. Based on x=7, with no populational variability the Iranian materials of A. desertrum belongs to only tetra- and that of A. cristatum to tetra- and hexa-ploid levels. Regarding the karyotyping features these two species are very similar. Our SSR results showed that while the highest genetic diversity appeared among the populations belonging to northern parts and the lowest within the eastern populations there no clear distinction between the two species. The northern, north-western and western populations showed very similar genetic diversity. Based on the results of this study it is suggested that A. desertrum to be lumped as a subspecies into A. cristatum; the only Agropyron species in Iran with eight infra-specific taxa in.
2-5.
Genome constitution of Hystrix komarovii (Poaceae: Triticeae)
Zhang H-Q, Fan X, Huang Y, Sha L-N, Zhou Y-H
Triticeae Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China; Key Laboratory of Crop Genetic Resources and Improvement, Ministry of Education, Sichuan Agricultural University, Yaan 625014, Sichuan, China
The genome constitution of Hystrix komarovii was examined by genomic in situ hybridization (GISH) and genome-specific RAPD assay. GISH of Pseudoroegneria spicata (St genome) and Hordeum bogdanii (H genome) probes confirmed the presence of the StH genomes in H. patula, but did not identified its presence in H. komarovii. The Ns and Ee genomic probes from diploids Psathyrostachys juncea and Lophopyrum elongatum did not discriminate the genomes of H. patula, whereas they produced strong hybridization signals on the two genomes of H. komarovii. Results of genome-specific RAPD assay were comparable with those of GISH, except that the Ee- and Eb-genome-specific RAPD bands were absent in H. komarovii. The results indicated that H. komarovii might contain the Ns and Ee genomes, but lack the StH genomes. Further cytological data are required for verifying the genome constitution of H. komarovii.
2-6.
A biosystematic study in Aegilops neglecta – Ae. columnaris species complex
Ohta S, Fujita Y, Maesaka Y, Hattori M, Iwasaki R
Department of Bioscience, Fukui Prefectural University, Fukui, Japan
Aegilops neglecta Req. ex Bertol. consists of tetraploid and hexaploid cytotypes. Ae. columnaris Zhuk. is a tetraploid species closely related to the tetraploid cytotype of Ae. neglecta. The aim of the present study is to clarify the morphological variation and cytogenetic relationship in tetraploid Ae. neglecta and Ae. columnaris. Spike characters of 131 accessions of tetraploid Ae. neglecta and 12 of Ae. columnaris collected from the whole distribution area were measured. The morphological variation was continuous but showed a significant correlation between the two characters, the length of rachis internodes at the base of spike and the shape of empty glumes: accessions with longer basal rachis internodes had circular glumes while those with shorter basal rachis internodes had elliptic glumes. Ae. columnaris was the extreme of the latter, while Ae. neglecta var. contorta was the extreme of the former. The variation showed a clear geographical cline from west to east, or coast to inland. Furthermore, 32 accessions of tetraploid Ae. neglecta (9 of var. contorta and 23 of the other varieties) and 8 of Ae. columnaris were artificially crossed with each other within and among taxa and geographical regions to clarify the degree of reproductive isolation barriers. Intraspecific F1 hybrids, especially those in Ae. neglecta, showed wide-ranged fertility. The fertility in interspecific neglecta-columnaris F1 hybrids was not lower than that in intervarietal hybrids in Ae. neglecta. From these results, we conclude that tetraploid Ae. neglecta and Ae. columnaris make a species complex in a biological species with a wide genetic variation.
2-7.
Using discriminant analysis to identify genomic groups within the perennial Triticeae
Rollo J1, Jacobs SWL2, Rashid A3, Barkworth, ME1
1Intermountain Herbarium, Dept. of Biology, Utah State University, Logan, Utah, 84322-5305, USA
2National Herbarium of New South Wales, Mrs Macquaries Road, Sydney, New South Wales, 2000 Australia
3University of Peshawar Botanic Garden, University of Peshawar, Peshawar, Northwest Frontier Province, Pakistan
As part of an investigation into the diagnosability of genomic groups, we scored 58 quantitative characters on 219 specimens of perennial Triticeae with solitary spikelets in their genomic group. The specimens represented 78 taxa and 13 different genomic groups. When all the specimens were used in the analyses, 98% of the specimens were placed in the correct genomic group. Jackknife analysis was less successful. In Jack knife analysis, one specimen is excluded from the analytical phase and used as test case. Using this procedure, only 80% of the specimens were placed in the correct genomic group. There were some subsets that for which identification was reasonably successful. For instance, the Australasian species, all of which contain the W genome, were successfully distinguished from the non-Australasian species 94% of the time. When the analysis was restricted to the StH and StY genomic groups (Elymus sensu stricto and Roegneria, respectively), the jack knifed success rate was 73%. It dropped to 69% if the StYP (Kengyilia) and P (Agropyron) groups were included.
2-8.
Geographical distribution patterns of morphological characters in cultivated barley (Hordeum vulgare L.) inferred from botanical varieties
Knüpffer H
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany
The German Genebank at IPK has a collection of the genus Hordeum with 23,000 accessions, it is the 6th largest barley collection worldwide. Most of these belong to cultivated barley, Hordeum vulgare. Many of the barley accessions represent landraces or traditional cultivars. Based on a number of morphological spike characters, the accessions are being grouped into botanical varieties using the infraspecific classifications of Mansfeld (1950) and Lukyanova et al. (1990) distinguishing 192 and 218 varieties, respectively. Such a classification of genebank material is very useful for purposes of the maintenance of the collection. The characters used are, among others, kernel row number (2 or 6), karyopsis type (covered, naked), spike density (lax, dense, very dense), glume width (narrow, broad), awn length and hoodedness (long, short, hooded, awnless), spike colour (yellow, various colours), awn roughness (rough, smooth), kernel colour (amber, various colours – considered for naked barleys only). In addition, growth habit (spring, winter, intermediate) is also considered.
Using various external data sources with pedigree and breeding information, named cultivars were classified into “recent” and “traditional” (released till 1940). Forty-seven countries, or groups of smaller countries, were found to be represented by at least 15 accessions of landraces or traditional cultivars with known botanical variety. The names of botanical varieties were translated into the corresponding character combinations. Thus it is possible to show the distribution of some morphological characters by countries (or groups of countries).
2-9.
Intraspecific variation of chloroplast DNA in Aegilops speltoides
Mori N1, Watatani H1, Ishii T2, Kondo Y1, Kawahara T3, Nakamura C1
1Lab. Plant Genetics, Graduate School of Agricultural Science, Kobe University, Kobe, Japan
2Lab. Plant Breeding, Graduate School of Agricultural Science, Kobe University, Kobe, Japan
3Plant Germplasm Institute, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
Chloroplast DNA polymorphism was investigated to clarify the intraspecific variation in Aegilops speltoides Tausch. Allelic diversity at 24 microsatellite loci was surveyed using 92 accessions of Ae. speltoides collected across its natural distribution area. Sixty-three accessions of wild emmer wheat (Triticum turgidum ssp. dicoccoides), six accessions of domesticated emmer wheat (T. turgidum ssp. dicoccum), and T. aestivum cv. Chinese Spring were used as the references. Average number of alleles in Ae. speltoides (4.00) was larger than that in the wild emmer wheat (3.26). Estimated diversity index (H) was also slightly larger in Ae. speltoides (0.37) than that in the wild emmer wheat (0.33). All 24 microsatellite loci except for WCt20/21 were highly polymorphic and 92 accessions of Ae. speltoides were classified into 80 plastotypes. Forty-eight and two plastotypes were identified in the wild and domesticated emmer wheat, respectively. Phylogenetic analysis revealed that all the plastotypes found in emmer wheat and those found in Ae. speltoides were clustered into two separate plastogroups. This result indicated the clear differentiation of plastom between the wild emmer wheat and Ae. speltoides. The 80 plastotypes found in Ae. speltoides were further grouped into two subgroups, suggesting an intraspecific differentiation within Ae. speltoides.
2-10.
Morphological variations of spike and the geographical distribution of subsection Emarginata species, genus Triticum-Aegilops, close wild relatives of wheat
Ohta A1, Kawahara T2, Yamane K1
1Grad. Sch. Bio. Env. Sci., Osaka Pref. U., Japan
2Grad. Sch. Agr., Kyoto U., Japan
To enhance use of wild relatives of cultivated plants as a genetic resource, it is important to understand the genetic diversity and its genetic background. We investigated morphological variations of spike of four subsection Emarginata species, Aegilops searsii, Ae. bicornis, Ae. longissima and Ae. sharonensis. A total of 102 accessions from the collection maintained at the Laboratory of Crop Evolution, Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University were grown at the experiment field of Kyoto University, Japan in 2007-2008. The result showed a correlation between the morphological variations of spike (spike length, awn length, spikelet density) and the geographical distribution (Mediterranean site or inland site). Our result showed the contradiction between the morphological similarity of spike observed in this study and previous studies of the molecular phylogenetic relationships among 4 species. It suggested the possibility of convergent evolution of spike morphologies in subgenus Emarginata. In order to clarify the genetic factor that caused the differences in the spike morphologies, we examined the relationships between the spike morphologies and climate of collection sites of the Emarginata accessions. Climate data was obtained from Worldclim database. As a result, a cline of spike morphology was found along geographical gradient of climate change. Our results suggested that winter temperature affects spike morphologies.
2-11.
The regulatory network underlying the six-rowed spike in barley
Pourkheirandish M, Komatsuda T
National Institute of Agrobiological Sciences (NIAS), Plant Genome Research Unit, Kan-non-dai 2-1-2, Tsukuba, Ibaraki 305 8602, Japan
Many important phenotypic traits are controlled by networks of genes. The architecture of the Hordeum spike, which is characterized by the presence of three spikelets at each rachis node, is unique among the Triticeae. Because the two-rowed spike is universal in wild barley, it is likely to be the ancestral form, and that the six-rowed spike arose by natural mutation during the domestication process. Recessive alleles present at five independent genes are capable of producing a six-rowed spike. The six-rowed spike 1 locus (vrs1 or Vrs1) is located on chromosome arm 2HL. Wild barleys and two-rowed cultivars carry the dominant Vrs1 allele, while all six-rowed cultivars carry the recessive vrs1 allele. Over 90 induced vrs1 mutants have been generated from two-rowed barley, supporting the notion that cultivated six-rowed barley was derived from a two-rowed progenitor. Recessive mutations (vrs2, vrs3, vrs4 and vrs5, which is synonymous with int-c) have been induced at the four other loci, located on, respectively, chromosome arms 5HL, 1HL, 3HL and 4HS, but no cultivated six-rowed type is determined by a variant at any of these genes. The recessive alleles at these four loci all act to enhance the development of the lateral spikelets, although the degree of this enhancement depends upon the spikelet's position along the spike. Vrs1 encodes a member of the homeodomain-leucine zipper (HD-ZIP) I class of transcription factors, which binds to its gene target as a homo- or a heterodimer. The six-rowed spike genes may form a gene network involved in the control of lateral spikelet development. The expression of Vrs1 in vrs2, vrs3, vrs4 and vrs5 backgrounds should be informative as to the identity of some of the genes acting upstream of Vrs1 in this network.
2-12.
The barley vrs1 gene evolved from duplication of a well-conserved HD-Zip I-class homeobox gene in the Poaceae
Sakuma S1,2, Pourkheirandish M1, Matsumoto T1, Koba T2, Komatsuda T1
1National Institute of Agrobiological Sciences (NIAS), Plant Genome Research Unit, Kan-non-dai 2-1-2, Tsukuba, Ibaraki 305 8602, Japan
2Graduate school of Horticulture, Chiba University, 648 Matsudo, Chiba 271 8510, Japan
Spike of cultivated barley (Hordeum vulgare ssp. vulgare) is composed of three spikelets at each rachis node. In two-rowed barley, the central one is fertile and the two lateral ones are sterile, whereas in the six-rowed type, all three are fertile. This characteristic is determined by the allelic constitution at the six-rowed spike 1 (vrs1) locus on the long arm of chromosome 2H, with the recessive allele (vrs1) being responsible for the six-rowed phenotype. The Vrs1 (HvHox1) gene encodes a homeodomain-leucine zipper (HD-Zip) transcription factor which is common in plant kingdom. Here we show that the Vrs1 gene evolved in the Poaceae via a duplication, with a second copy of the gene, HvHox2, present on the short arm of chromosome 2H. Micro-collinearity and polypeptide sequences were both well conserved between HvHox2 and its Poaceae orthologues, but Vrs1 is unique to the barley tribe. The Vrs1 gene product lacks a motif which is conserved among the HvHox2 orthologues. A phylogenetic analysis demonstrated that Vrs1 and HvHox2 must have diverged after the separation of Brachypodium distachyon from the Pooideae. The loss of Vrs1 function both in natural variants, and in many induced mutants (including the total deletion of the gene) indicates that Vrs1 is non-essential for plant growth and development. From these analyses it is suggested that Vrs1 arose following the duplication of indispensable gene HvHox2, and acquired its new function during the evolution of the barley tribe.
2-13.
Intraspecific variation in leaf shape-related traits in a wild einkorn wheat species Triticum urartu Thum.
Morihiro H, Takumi S
Graduate School of Agricultural Science, Kobe University, Rokkoda-cho 1-1, Nada-ku, Kobe 657-8501, Japan
Einkorn wheat contains three species; cultivated Triticum monococcum and wild T. boeoticum and T. urartu. Triticum monococcum was domesticated from T. boeoticum. Triticum urartu is an A genome donor for the polyploid species of Triticum. Our previous study showed that both nuclear and chloroplast genomes of T. urartu were clearly differentiated from those of T. monococcum and T. boeoticum, and that the T. urartu accessions were classified into two major haplogroups based on their chloroplast DNA variations. In this empirical study, we analyzed intraspecific variation of eight leaf shape-related traits using 30 accessions of T. urartu, 2 T. boeoticum accessions and 2 T. monococcum accessions. Principal component analysis of the 8 leaf shape traits indicated that two subgroups were diverged in the T. urartu population and that this intraspecific differentiation was corresponding to the two haplogroups based on the chloroplast DNA variations. The subgroup diversification of T. urartu was largely caused by leaf length. Significant difference of the flag and its lower leaf length was observed between the two subgroups. One subgroup with the short leaf length contained accessions mainly collected in Armenia and Lebanon, and another with the long leaf length included a lot of accessions in Iran and Turkey. On the other hand, four or five different clusters could be divided in the T. urartu accessions revealed by the AFLP analysis of total DNA. Interestingly the subgroup diversification was not consistent with the clusters based on nuclear DNA variations.
2-14.
Allopolyploidy of the Hordeum murinum complex indicated by a nucleotide sequence of cMWG699
Tanno K1,2,3, Bothmer von R4, Yamane K5, Takeda K1, Komatsuda T2
1Research Institute for Bioresources, Okayama University, Japan
2National Institute of Agrobiological Sciences, Japan
3Yamaguchi University, Japan
4Swedish University of Agricultural Sciences, Sweden
5Osaka Prefectural University, Japan
Hordeum murinum is one of the most widely distributed plant in the genus Hordeum and is well known as a nuisance weed in the world temperate zoon. This species is composed of three subspecies with three ploidy levels, namely ssp. glaucum (2x=14), ssp. murinum (4x=28) and ssp. leporinum (4x=28 and 6x=42). Their morphological features, however, resemble each other, they are frequently collectively referred to as the “murinum complex”.
Many cytological studies suggest allopolyploidy nature of the murinum complex while autopolyploidy is also suggested by an inter-specific study. The present study is aim to clarify the allo- vs. auto- polyploidy status of the murinum complex based on molecular phylogenetic analysis, especially focusing on nucleotide variations and divergence of the polyploid genomes.
A single nuclear DNA locus, cMWG699, was analyzed, this nucleotide sequence has been used in a series of phylogenetic studies of the Hordeum species. PCR-RFLP analysis with HhaI and SspI for 80 H. murinum accessions revealed polymorphism between diploid and polyploids, and double-digestion with the two enzymes showed polymorphism between tetra- and hexa-ploids. Nucleotide sequence of sub-clones clearly showed allopolyploid nature of the polyploid murinums.
The sequence study clearly indicated that one donor of the hexaploid was ssp. glaucum (2x) as has been suggested by cytological studies. The other donors were not identified for their original diploid species nevertheless they were at least within the diversity of the genus Hordeum. Regarding the two subspecies of tetraploid, nucleotide sequences of 4x ssp. murinum and 4x ssp. leporinum were highly similar, further study using the different loci is necessary for the classification of the two subspecies.
2-15.
The variation of SSR profiles in wild and cultivated barley
Turuspekov Y, Abugalieva S
Institute of Plant Biology and Biotechnology, Almaty 050040, Kazakhstan
Nineteen SSR primers were analyzed to order to assess the genetic diversity of 68 barley cvs. and 13 wild populations of H. spontaneum K. from Israel, Turkmenistan, and Kazakhstan. In total 254 alleles from 22 SSR loci were revealed using 6% polyacrilamide gel electrophoresis. The results are following: a) amount of polymorphism for wild barley (He=0.71) was higher than for cultivated (He=0.63); b) higher amount of genetic variation of cultivated cvs. from Kazakhstan in compare with European samples, and it was within a range of genetic diversity for wild barley; c) of the total genetic diversity of Hordeum, 69.83% was within populations, 9.28% between populations within a species, and 20.89 between species; d) the structure of genetic diversity for H. spontaneum was 36.10% within populations, 50.16% between populations of a region, and 13.74% between regions; e) the level of polymorphism did not positively correlated with sample size of wild populations; and f) the genetic distance of wild populations did not relate with their geographic distance. Results confirmed high potential of SSR markers for genetic diversity analysis and efficient identification of barley genotypes.
2-16.
A novel source of germplasm for the development of branched ear wheat
Aliyeva AJ, Aminov NKH
Department Cytogenetics, Institute of Genetic Resources, 155, Azadlig Ave., AZ 1106, Baku, Azerbaijan
A novel source to produce branched-ear trait in durum wheat was developed from the complex hybridization among Triticeae species. After a synthetic hexaploid wheat (T. durum x Ae. squarrosa, 2n=42) was crossed with rye (S. segetale Roshev, 2n=14), hybridization between the three generic incomplete amphidiploid (Aegilotriticale, 2n=42) and common wheat (T. aestivum cv. Chinese Spring, 2n=42) was made. A line ‘171ACS’ obtained from the latter hybridization was further crossed with durum and common wheat accessions. All the resulted F1 plants had normal spikes. In F2 generation, all populations derived from the crosses between durum wheats and a line ‘171ACS’ segregated for normal and branched ears. The branched ear was not observed from the crosses between ‘171ACS’ and common wheat. The phenotype of branched ears differed from known branched ear wheat i.e. T. turgidum L. or T. vavilovii Jakubz, but closer to the morphology of T. vavilovii. Each spikelet showed hierarchical non-terminated structure. The branched-ear trait is controlled by a single recessive gene estimated by the above Mendelian segregation of the F2 population and the backcross population of the same cross. The result indicated that branched-ear development was suppressed by the factor derived from the D-genome of common wheat. To confirm the existence of this factor in D-genome, crossings between '171ACS' and synthetic tetraploid wheat (T. urartu x Ae. tauschii, AADD) are underway.
2-17.
Isolation and molecular characterization of three novel HMW glutenin subunits from Aegilops tauschii
An X1,2, Wang D2, Yan Y1
1Key Laboratory of Genetics and Biotechnology, College of Life Science, Capital Normal University, Beijing 100037, China
2The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
High molecular weight glutenin subunits (HMW-GSs) exhibit abundant allelic variations in wheat related species. Three novel HMW-GSs from Aegilops tauschii were identified by SDS-PAGE, RP-HPLC and MALDI-TOF-MS. Their complete coding genes were amplified and cloned with AS-PCR primers, named as Glu-1Dx1.6t Glu-1Dx3t and Glu-1Dx5.2t, respectively. The primary structure of the three subunits was highly similar to that of the previously reported Glu-1Dx subunits in wheat, but also displayed unique features. Particularly, Glu-1Dx5.2t subunit contained an extra cysteine residue in the repetitive domain in addition to the four conserved cysteine residues commonly found in the N-terminal domain of homoeologous x type subunits. The structural features of Glu-1Dx5.2t resemble closely those of the good quality subunit 1Dx5 that is frequently used in breeding wheat varieties with superior bread making properties. Based on this and previous studies, the origin and evolution of 1Dx5 subunit in common wheat are discussed.
2-18.
Analysis grain characteristics of tetraploid wheat gene pool to diversify genetic background of durum wheat
Taguchi J1, Kiribuchi-Otobe C2, Matsunaka H2, Ban T1
1Kihara Inst. Biol. Res., Yokohama City U., Yokohama, Kanagawa 244-0813, Japan
2NICS, Tsukuba, Ibaraki 305-8518, Japan
It has been not cleared what kind of grain characteristics are related to quality of tetraploid wheat gene pool. This study we examined genetic diversity for several traits associated with pasta making qualities among tetraploid wheat germplasms conserved in KIBR collection.
We analysed 96 acessions of tetraploid wheat including Triticum timopheevi, T. turgidum (convs. durum, turgidum, dicoccum, orientale, pyramidale and carthlicum) and D genome chromosome substitution lines of Langdon durum for each homoeologous A and B chromosomes. We identified genotype of HMW-gulutenin subunits by SDS-PAGE, genotype of NAC gene which is considered to improve protein and Fe, Zn content in wheat grains (Uauy et al. 2006) by PCR, measured seed protein content by Dumas combustion analysis (rapid-NR, elementar) for N/Protein, amylase content with AutoAnalyser (Technicon), and content of grain yellow pigment (GYPC) with absorption spectrophotometer. Regarding with genotype of HMW-gulutenin subunits, T. timopheevi and T. turgidum conv. turgidum had specially Glu-B1j, Glu-B1h allele, respectively. T. turgidum conv. turgidum had lower protein content and T. timopheevi had higher protein content relative to T. turgidum conv. durum. Amylose content was about 27 % in almost of all line, but only T. timopheevi showed 22%. Regarding with NAC gene, T. turgidum conv. turgidum and T. timopheevi carried NAM-B2 addition to NAM-A1 and NAM-B1 allele which T. durum carried. About GYPC, T. timopheevi contain 1.4 times content relative to T. turgidum conv. durum. The D genome chromosome substitution lines of Langdon durum showed broad range of variability on the traits. This study showed valuable diversity of the pasta making traits to improve durum wheat with tolerance to biotic and abiotic stresses from the tetraploid wheat gene pool. This work was supported to use KIBR germplasms contributed by NBRP project.
2-19.
Study of diversity and relationships of the D genome species of Aegilops-Triticum from Iran
Bordbar F[1,2], Rahiminejad MR2, Saeidi H2, Blattner FR1
1Leibniz Institute of Plant Genetics and Crop Research (IPK), D-06466 Gatersleben, Germany
2Department of Biology, University of Isfahan, Isfahan, Iran
As hexaploid bread wheat (Triticum aestivum) contains a D genome, Aegilops species bearing the D genome can be used as potential sources of useful alleles to be incorporated into cultivated wheat. These Aegilops species posses a D genome component derived from their diploid progenitor A. tauschii. We studied 76 accessions of D genome species of the Aegilops-Triticum complex from different parts of Iran together with 39 accessions of A. ventricosa, A. vavilovii, T. aestivum and A. juvenalis from other geographic regions. For estimation of genetic diversity within and between species, 24 highly informative D genome-specific microsatellite markers were used, together with sequences from nuclear rDNA spacers and a chloroplast intergenic spacer region.
Analyses of SSR diversity showed A. tauschii to possess a wide range of alleles, as is evident from high PIC values. Phylogenetic trees indicate A. tauschii to be an old lineage with high genetic differentiation of the populations found in Iran. All our results revealed A. tauschii to consist of two different gene pools. Some of the accessions are more closely related to A. cylindrica, while other accessions of A. tauschii clustered with the accessions of T. aestivum. The analyzed accessions of A. cylindrica showed low PIC values and very low genetic distances compared to the other species. The results of this study confirmed that the D genome of A. tauschii, A. cylindrica and T. aestivum are different from the D genomes found in A. crassa, A. juvenalis, A. vavilovii and A. ventricosa, and indicate a cryptic taxon within A. tauschii, as the two groups defined by molecular methods are not in accord with the already described subspecies of this taxon.
2-20.
Estimation of quality of Triticum durum Desf. wheat on the basis of gliadin and glutenin characterisation
Gregová E1, Medvecká E2, Šramková Z3, Mihálik D1
1Research Institute of Plant Production, Bratislavská cesta 122, 921 68 Piešťany, Slovak Republic
2Constantine the Philosopher University, Faculty of Natural Sciences, Department of Chemistry, Tr. A. Hlinku 1, 949 74 Nitra, Slovak Republic
3Slovak University of Technology in Bratislava, Faculty of Chemical and Food Technology- Institute of Biochemistry, Nutrition and Health Protection, Radlinského 9, 812 37 Bratislava, Slovak Republic
The aim of the studies was the electrophoretic characterization gliadin and glutenin proteins and evaluation of in kernels of Triticum durum Desf.. All 56 accessions originating from different geographical areas of Europe were evaluated for high molecular weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW) composition using SDS-PAGE and A-PAGE. The data indicated the prevalence of the null allele (95%) and 1 subunit (5%) at the Glu-1A and four alleles, namely 6+8 (33%), 7+8 (32%), 13+16 (18%) and 20 (9%) represented at the Glu-1B. Protein subunit Glu-1A1 was correlated positively with improved dough strength as compared to subunit null. On the chromosome Glu-1B subunit 6+8 was associated with slightly stronger gluten type than 7+8 and 13+16, while subunit 20 was associated with weak gluten properties. On the basis of electrophoretic separation of gliadin fraction it was found that 49 genotypes contained γ-45, 1 genotype γ-42 and 6 genotypes another. Cultivars having the low molecular weight (LMW) glutenin allele LMW-2 (or gliadin band γ-45) generally gave stronger gluten than lines with allele LMW-1 (or gliadin band γ-42). The combined better alleles at Glu-B1 (coded bands 6+8, 13+16, 7+8) and Glu-3 (patterns LMW-2) showed linear cumulative effects for dough strength. These results could provide a more complete understanding of the studied collections diversity on high molecular subunits and it will be useful to breeders who now possess a tool to formulate crosses by choosing varieties with appropriate characters.
2-21.
Genetic variability in bread wheat (Triticum aestivum L.) of Slovakia based on polymorphism for high molecular weight glutenin subunits
Mihálik D1, Šramková Z2, Medvecká E3, Horevaj V4, Šliková S1
1Research Institute of Plant Production, Bratislavská cesta 122, 921 68 Piešťany, Slovak Republic
2Slovak University of Technology in Bratislava, Faculty of Chemical and Food Technology- Institute of Biochemistry, Nutrition and Health Protection, Radlinského 9, 812 37 Bratislava, Slovak Republic
3Constantine the Philosopher University, Faculty of Natural Sciences, Department of Chemistry, Tr. A. Hlinku 1, 949 74 Nitra, Slovak Republic
4Hordeum, s.r.o., Nový Dvor 1052, SK-92521 Sládkovičovo, Slovak Republic
The genetics and biochemistry of high molecular weight glutenin subunits (HMW GS) in wheat is very well known now. The results have shown that three loci coding for HMW GS are highly polymorphic in nature without being influenced by the environment. Forty-six samples of common wheat varieties registered in the period 1976 – 2008 in Slovakia, kept in the collection of the Genebank Piešťany, were surveyed to determine their high molecular weight glutenin subunits as separated by polyacrylamide gel electrophoresis (SDS-PAGE). The high-molecular-weight (HMW) glutenin subunits, group of storage proteins of wheat, are encoded by genes at three complex loci (Glu-A1, Glu-B1 and Glu-D1) located on the long arms of the homoeologous chromosomes 1A, 1B and 1D respectively. Total proteins where extracted from crushed half grains and separated according to the method of Wrigley (1992). The HMW glutenin subunits were classified according to the numbering system of Payne and Lawrence (1993). A total number of 10 alleles were detected at all Glu-1 loci, 3 belonging to Glu-A1, 5 to Glu-B1 and 2 to Glu-D1 locus, respectively. The allele most strongly associated with good quality, Glu-D1 subunits 5+10, was present in 82.6% of the studied wheat genotypes. The most common banding patterns were Null (on chromosome 1A), 7+9 (on chromosome 1B) and 5+10 (on chromosome 1D), respectively. The results of the study showed that 44 of the analysed accessions were found to be homogenous. The other 2 consisted of at least 2 protein lines (phenotypes).
2-22.
Multiplex quantitative analysis for trichothecene genes expression of Fusarium graminearum causing head blight on wheat spikes
Miyazaki T, Ban T
Kihara Inst. Biol. Res., Yokohama City U., Yokohama, Kanagawa 244-0813, Japan
Fusarium graminearum attacks spike of Triticeae species and causes head blight (FHB) disease. It results in contamination of trichothecene mycotoxins in the graitns as a global threat of food production and hygiene. Wheat resistance level to mycotoxin contamination varies among genotypes, however it remains incompletely understood whether resistance to fungal invasion produces secondary effect or specific genes works on low level accumulation of the mycotoxins. We applied multiplex quantitative gene expression analysis for the trichotecene production (Tri) genes of F. graminearum to reveal effective wheat genotypes and germplasms to reduce mycotoxin production. We designed reverse chimera primers, which consisted of 3’ side specific sequence of Tri genes and universal primer sequence tail, to synthesize single strand cDNA from total RNA. Forward chimera primers with a universal tail designed to amplify different size of each Tri gene added for 1st step PCR up to 3 cycles to quantify the expression. Then, universal primer sets, which forward primer was labeled with fluorescent at 5’ end, amplified fragments around 100 to 600bp length to be detected by the capillary sequencer (Genome Lab GeXP, BECKMAN COULTER). As a result, Tri genes (Tri5, Tri6, Tri8, Tri10, Tri11) expression was detected from 1.0 mg lyophilized hypae cultivated in a liquid medium and correlated with final myctoxin production level by means of ELISA. Likewise, we could evaluate of Tri genes expression level from a wheat spike which infected by F. graminearum at flowering time. All of the Tri genes expression was promoted in a FHB susceptible wheat (Gamenya) more than resistance one (Sumai 3). We chose wheat genes associated with FHB resistance to introduce the multiplex PCR to examine interaction between wheat and F. graminearum gene expression. This method is cost-effective to the microarray analysis and high throughput then the real time PCR method.
2-23.
New tools for the accessibility of the Spanish barley core collection
Igartua E2, Molina-Cano JL1, Gracia MP2, Casas AM2, Moralejo M1, Ciudad FJ3, Lasa JM2
1IRTA, Av. Rovira Roure 191, E-25198 Lérida, Spain
2EEAD-CSIC, Avda Montañana 1005, E-50059 Zaragoza, Spain
3ITACyL, P.O. Box 172, E-47071 Valladolid, Spain
The Spanish Barley Core Collection (SBCC) was conceive as a resource for research, and was assembled as a representative sample of the genetic variability present in the collection of over 2000 accessions held at the National repository for plant genetic resources (CRF-INIA). The SBCC consists of mostly inbred lines derived from landraces, adapted to Southern European conditions. It is a unique germplasm resource, holding a great amount of diversity, originated in an abiotically stressed environment, isolated from mainstream gene pools, already studied to an unparalleled level among European landraces. It has been thoroughly studied for over 30 agronomic, disease resistance, morphological, and quality traits, revealing great potential to become a source of novel alleles for adaptive traits and disease resistance. Its adaptation to Mediterranean climates also anticipates the existence of drought tolerance traits, as a result of adaptation to the prevailing semi-arid conditions. Most information has been made available to the scientific community through the web site http://www.eead.csic.es/EEAD/barley/index.php, which includes passport data, evaluation data for morphological, agronomical and disease resistance traits. These data have been also summarized in a recent book published by the Spanish Institute for Agri-Food Research and Technology (in English): Spanish Barley Core Collection, JM Lasa (coordinator), Monografías INIA: Serie Agrícola, 25-2008 (ISBN 84-7498-526-9).
2-24.
Triticum species in Georgia: diversity, conservation, and taxa of special interest
Mosulishvili M1, Maisaia I2, Shanshiashvili T2, Akhalkatsi M1
1Ilia Chavchavadze State University. 32 Chavchavadze Ave., 0179 Tbilisi, Georgia
2Tbilisi Botanical Garden & Institute of Botany. 1 Botanikuri str., 0105 Tbilisi, Georgia
Georgia is one of the centers of evolution for many cereal crops and is rich with genetic resources of crop wild relatives. Earlier investigators of Georgian crop plants include N. Vavilov, who gave great attention to the Transcaucasus in his research and visited Georgia only 16 times. Georgian scientists V. Menabde and A. Dekaprelevich studied phylogenetics of Georgian endemic wheat species. The first evidence of farming civilization discovered in Georgia dates back to the Mesolithic period. Wheat, barley and millet were already cultivated in Georgia in the Eneolithic and Early Bronze periods. Anterior Asia is the native place of 12 wheat endemic (s.str.) species, 8 of them originate from the Transcaucasus, and among these endemics of the Transcaucasus, 5 species (s.str.) originate from Georgia. Georgian endemic taxa are valuable genetic material for modern selection. Particularly, tetraploid (Triticum timopheevii) and hexaploid (T. zhukovskyi) wheats are characterized by high resistance to diseases. T. carthlicum is characterized by immunity to diseases, short growing period and is cold-resistant. In the past this species was widely spread in Georgia in the Javakheti (Upper Kartli) region. Georgia is very rich in agricultural plants and it is a country of unique diversity of old varieties and landraces of wheat and other cereals. Many varieties were lost during the last 20 years but some were saved in seed banks and farms; 14 species of wheat, 144 varieties and 150 forms were registered till 1940-50s. The number of varieties and forms has been catastrophically decreased and suffered rapid genetic erosion. These taxa must be rediscovered and conserved both ex situ in seed and living plant collection and in situ in their original habitat in Georgia.
2-25.
Assessment of genetic diversity among Azerbaijan barley genotypes (H. vulgare L.) based on Hordein alleles
Geraybeyova N, Sadiqov H, Rahimova O, Sadiqova S, Karimov A, Mammadova N, Babayeva S, Abbasov M
Genetic Resources Institute of ANAS, Baku, Azerbaijan
Although H. vulgare ssp. spontaneum (C. Koch) Thell is prevalent plant in Azerbaijan widely distributed almost all over the country, precise and detailed information on genetic diversity is lacking and little is known regarding population structure and geographic differentiation of this plant. Therefore, we analyzed H. spontaneum populations collected from different regions of Azerbaijan with the objectives: to get information on genetic polymorphism degree within H. spontaneum populations, as well as within revealed by us H. ishnaterum Coss. variety and to study differences between spontaneum and vulgare varieties based on hordein electrophoretic components. The electrophoretic analysis of 61 barley genotypes was carried out according to modified method of F.A. Poperelya et al. (2001). In an electrophoretic survey of eight H. spontaneum and one H. ishnaterum Coss. genotypes high polymorphisms degree was observed for hordein coding locus - HRD A and low polymorphism for HRD B and HRD F loci. Comparison of electrophoregram for hordein coding loci in spontaneum and 52 vulgare (50 local and 2 introduced) genotypes revealed 100% polymorphism degree within HRD A -5, 6, 8 and from 6.2 to 8.8% polymorphism within HRD A 1, 2 loci. The polymorphism percentage for HRD B 1, 2, 3, 4 was within the range of 6.2-62.5%. Considerable genetic variation was noted for HRD F1 ranging from 6.2 to 100%, whereas for HRD F9 and HRD F 4, 6, 7 it varied from 69.2 to 81.8% and from 6.2 to 15.3% respectively. The results proved that in studied genotypes the polymorphism was observed mainly in HRD A and partly in HRD B hordein blocks.
2-26.
Study of sequence polymorphism and genetic diversity of sucrose-phosphate synthase genes in bread wheat and its A, B and D genome progenitors
Sharma S, Röder MS
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466 Gatersleben, Germany
Sucrose phosphate synthase (SPS) is a main regulatory enzyme involved in sucrose biosynthesis pathway in many crop plants, including wheat. Present study was designed to study the structure of SPS genes in wheat. SPS is organized in several gene families in the wheat genome. Genome specific primers for SPS gene family II were designed for SPS genes using exon anchored primers that could amplify one or more introns, which are supposed to be more polymorphic than the exons. Each of the individual SPS loci of the three wheat A, B and D genomes was studied. Approximately, 4kb of each genome could be compared and analysed. Detailed investigation of introns and exons was conducted. Sequences alignment was conducted to search for the SNPs present in each of the three genomes. So, developed SPS genome specific primers are being used for the PCR amplification in multiple accessions of T. aestivum, T. turgidum subsp. durum, T. urartu, Ae. speltoides and Ae. searsi. Sequence polymorphism, separately for each wheat species, will be grouped into distinct haplotypes. Genetic mapping and genetic diversity studies in common wheat, for SNP sites will also be conducted using either pyrosequencing or direct sequencing. Linkage disequilibrium analysis will also be performed.
2-27.
Composition of high-molecular-weight glutenin subunits in European wheats
Šliková S1, Šramková Z2, Gregová E1, Mihálik D1
1Research Institute of Plant Production, Bratislavská cesta 122, 921 68 Piešťany, Slovak Republic
2Slovak University of Technology in Bratislava, Faculty of Chemical and Food Technology- Institute of Biochemistry, Nutrition and Health Protection, Radlinského 9, 812 37 Bratislava, Slovak Republic
Seed storage proteins are considered to be usable markers for the studies of wheat genetic resources High-molecular-weight glutenin subunits (HMW-GS), present in the endosperm of hexaploid wheat (Triticum aestivum L.), are the main components of gluten, which is the main contributor to the rheological and bread-making properties of wheat flour. The objective of our study was to determine the HMW-GS composition of 57 wheat cultivars (T. aestivum L.) originated from six European countries, kept in the collection of the Genebank Piešťany. Protein profiles were examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Fourteen different alleles/allelic pairs were identified for the three glutenin loci studied, Glu-A1 (3), Glu-B1 (9) and Glu-D1 (2). All together, twenty-five glutenin patterns were detected for HMW-GS, occurring at various frequencies depending on country of origin. The most common allelic composition was 0 (Glu-A1), 7+9 (Glu-B1) and 5+10 (Glu-D1). Allele “null” was the most frequent (50-100%) subunit at Glu-A1 locus in wheat cultivars from all of the six countries. Consequently, the results also suggested that higher variability occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Rare alleles such as 7, 18, 20, 22 and 13+16 were found at Glu-B1 locus. The most frequent glutenin subunits encoded by Glu-D1 were 5+10, except cultivars originated from Italy and Great Britain, where subunits 2+12 were predominant (60-76, 9%).
2-28.
Genetic diversity within Triticum turgidum L. subsp. dicoccon (Schrank) Thell. (cultivated emmer) and its utilization in wheat breeding
Zaharieva M1, Dreisigacker S1, Crossa J1, Payne T1, Misra S2, Hanchinal RR3, Mujahid MY4, Trethowan R5
1CIMMYT, Mexico
2Agharkar Institute, Pune, Maharashtra, India
3University of Agriculture Sciences, Dharwad, Karnataka, India
4PARC, Pakistan
5Sydney University, Australia
Synthetic hexaploid wheat (SHW) obtained by crossing durum wheat (AB-genomes) and the wild relative Aegilops tauschii Coss. (D-genome) allowed a significant increase of cultivated wheat genetic diversity. Backcross derived lines (SBL), obtained by crossing SHW to elite bread wheat cultivars, have been already extensively used in wheat breeding. Higher genetic diversity was identified in cultivated emmer, Triticum turgidum L. subsp. dicoccon (Schrank) Thell., compared to durum wheat, suggesting that using emmer wheat to develop new SHW could provide access to new alleles and traits. Moreover, cultivated emmer has proven to be a valuable source of drought and heat tolerance and the first SBL generated using emmer wheat showed higher yield under drought-prone conditions in Mexico, Pakistan and India compared to those using durum wheat. These lines were found to be genetically diverse and distant from drought tolerant durum wheat based SBL and traditional bread wheat cultivars and breeding lines, confirming their interest as new sources of diversity. A research project has consequently been initiated to i) discover novel genetic diversity from emmer wheat, ii) develop new SHW from diverse emmer wheat accessions, iii) describe and compare diversity within emmer and durum based SHW to pyramid useful diversity and iv) develop new SBL by crossing the most diverse primary emmer based SHW to local elite bread wheats from different origins and breeding programs (CIMMYT, India, Pakistan, Australia).
As part of this project a collection of 300 accessions from different geographical regions covering the emmer wheat area of distribution was established at CIMMYT. Molecular characterization was carried out to understand the genetic structure of the species and identify accessions of interest. A subset of genetically diverse accessions was created to be further used for development of new pre-breeding wheat germplasm with enchanced drought and heat tolerance.
2-29.
Identification and mapping of QTLs for grain protein content in common wheat
Abugalieva S1, Abugalieva A1, Quarrie S2*, Turuspekov Y1
1Institute of Plant Biology and Biotechnology, NCB RK, Almaty, Kazakhstan
2John Innes Centre, Norwich NR47UH, UK
*Current address: Kraljice Natalije 39, 11000 Belgrade, Serbia
In this work we have identified quantitative trait loci (QTL) for grain protein content (GPC) in 95 doubled haploid (DH) lines of a mapping population derived from a cross between the common wheat genotypes Chinese Spring and SQ1 under the conditions of Southeast Kazakhstan. The GPC of DH lines was significantly different between rainfed and irrigated sites (P<0.05). In total, ten QTLs for GPC were found under the two treatments for moisture availability. Two QTLs for GPC under rainfed conditions were predicted to be novel in comparison with previous reports. The novel QTLs were mapped onto chromosomes 2BS and 5DL in the population grown under rainfed conditions. Closely-linked DNA markers were identified for the majority of mapped QTLs. The results could be implemented in a local breeding program for the improvement of wheat grain quality using marker-assisted selection. The study is the further contribution to our understanding of regions of the wheat genome that contribute to the control of GPC in common wheat.
2-30.
Investigations on yellow rust disease resistance by useful genes and markers in gene-rich regions on wheat chromosomes
Aydin Y1, Cabuk E1, Mert Z2, Akan K2, Bolat N3, Cakmak M3, Uncuoglu AA4
1Marmara University, Faculty of Science and Letters, Department of Biology,34722, Istanbul, Turkey
2Field Crop Research Institute, P.O Box: 226, Lodumlu, Ankara, Turkey
3Anatolian Agricultural Research Institute, P.O Box: 17, 26001, Eskişehir, Turkey
4The Scientific and Technological Research Council of Turkey (TUBITAK), Marmara Research Center (MRC), Genetic Engineering and Biotechnology Institute (GEBI), P.O Box: 21, 41470, Gebze, Kocaeli, Turkey
High density genetic linkage and physical maps are available for wheat and there are many markers and important genes, including Yr which controlling yellow rust disease resistance. One of the important gene-rich regions is present on the short arm of wheat homoeologous group 1 chromosomes. Resistant (İzgi01, Sönmez2001, PI178383) and susceptible (Aytın98, ES14, Harmankaya99) bread wheat genotypes were screened by 43 molecular markers located at gene rich regions on physical maps of wheat 1B chromosome. Polymorphic band patterns were obtained with 17 out of 43 markers in wheat genotypes. Furthermore, 7 Yr genes (Yr9-Yr15-Yr26-YrH52 on 1B, Yr7 on 2B, Yr17 on 2A, Yr18 on 7D) were used to investigate the DNA sequence differences between wheat genotypes regarding to yellow rust resistance. Based on the obtained results, polymorphic markers and Yr genes sequence differences between wheat genotypes were used to screen F2 populations from the cross of İzgi2001 x ES14, PI178383 x Harmankaya99 and Sönmez2001 x Aytın98 for bulk segregant analysis to find out molecular markers linked to yellow rust resistance. The presence of polymorphic markers that is associated with yellow rust resistance may significantly enhance the success of selection for yellow rust resistant genotypes in wheat breeding programs.
2-31.
Regulation of transformation efficiency in polyploid cereals by type and number of selection cassettes
Bińka-Wyrwa A, Orczyk W, Nadolska-Orczyk A
Plant Transformation and Cell Engineering Lab, Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland
The aim of this study was to optimize the process of selection and expression of transgenes in wheat and triticale plants obtained as the result of Agrobacterium-mediated transformation. We have had developed this transformation method for cultivars of both species. Currently we would like to explain the relationships occurring between the type and the number of expression cassettes represented by two selection genes: nptII and bar. Ten pGREEN (www.pgreen.ac.uk) vectors contained single or two in tandem arranged selection cassette(s) were constructed. The cassettes were driven by the same or different promoters: nos, 35S or Ubi1. All vectors were electroporated to A. tumefaciens, strain Agl1 and used for transformation of two cultivars of wheat (Kontesa, Torka) and one cultivar of triticale (Wanad). Selection efficiency in these cultivars transformed with 35S::nptII, nos::nptII, 35S::nptII/35S::nptII and nos::nptII /35S:: nptII ranged from 0 to 5%. It was higher in combinations of Kontesa and Wanad transformed with one copy of nos::nptII or 35S::nptII than with two copies of the same selection cassette. The second cultivar of wheat, Torka could not be transformed by any of three combinations containing nptII. The single plant was selected after transformation with doubled cassette of 35S::nptII. The same cultivars of wheat and triticale transformed with one or two cassettes of bar under nos, 35S or Ubi1 promoters gave the opposite results. Selection efficiency of Kontesa, Torka and Wanad transformed with 35S::bar, nos::bar, 35S::bar/35S::bar, nos::bar/35S::bar and 35S::bar/Ubi1::bar, nos::bar/Ubi1:: bar ranged from 0 to 6,2%. It was higher in combinations containing two copies of the bar cassette comparing with the combinations containing one copy. The highest selection efficiency was observed in combinations of Kontesa, Torka and Wanad transformed by two in tandem arranged bar cassettes driven by 35S and Ubi1 (35S::bar/Ubi1::bar) and/or nos and Ubi1 (35S::bar/Ubi1::bar).
Acknowledgements: This work was financed by the Polish Ministry of Science and Higher Education, grant PBZ-MNiSW-2/3/2006/31.
2-32.
Sequence variation of the 20th exon within PolA1 gene among Triticeae species
Buwan R1, Takahashi H1, Kato K2, Sato Y-I3, Komatsuda T4, Nakamura I1
1Graduate School of Horticulture, Chiba University, Matsudo, Chiba 271-8510, Japan
2Faculty of Agriculture, Okayama University, Okayama 700- 8530, Japan
3Research Institute of Human and Nature, Kamigyo-ku, Kyoto 602-0878, Japan
4National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
PolA1 gene encodes for the largest subunit of RNA polymease I complex and is present as a single copy per plant genome. Because sequences of the 20th exons were highly polymorphic in Oryza and Petunia, nucleotide sequences of PolA1 20th exons were analyzed for 13 Triticum, 14 Hordeum and 3 related species. Phylogenetic analysis of the sequences showed that Triticum and Hordeum species were distinctly separated into two major clades, except that SS genome species of Triticum were clustered with Hordeum species. Although Secale cereale was grouped into Triticum clade, Dasypyrum villosum belonged to Hordeum clade.
2-33.
High frequency and a wide spectrum of mutations in ‘BARONESSE’ barley fields
Cagirgan AMI1, Ullrich SE2, Ozbas MO1
1Department of Field Crops, Faculty of Agriculture., Akdeniz University, Antalya,Turkey
2Department of Crop and Soil Sciences, Washington State University, Pullman, WA, USA
Intensive agricultural systems increase agrochemicals' use such as fertilizer and pesticides. Lowering inputs such as no-till or reduced tillage is proposed as a way to sustainable agriculture by lowering costs and so increasing income from per unit area. However, the new production system generates problems of weeds and hence increases use of amount of herbicides to control weeds. In 1997, Cagirgan and Ullrich observed a high frequency and wide spectrum of mutations such as genetic male sterility, dense/lax spike, smooth awn, anthocyanin-less/more, eceriferum, etc in farmers' fields at Washington State, USA. Such huge variability would only be observable in a genetic background responding very well to a very efficient mutagen treatment. Considering this situation and assuming that the genetic background, Baronesse barley, is sensitive to mutate easily by any environmental/agrochemical force, we irradiated the cultivar twice by gamma rays of 100-300Gy, but almost nothing was selected at the station except some chlorophyll mutations in M2 generations of the two different irradiated materials although few mutants were selected by re-screening of the on-farm grown M3 populations yielding still low frequency and very narrow spectrum of mutations. Intensive herbicide use in the area may be the reason for the high frequency & wide spectrum of the mutants observed in Baronesse barley fields at Washington State.
2-34.
Characterization of growth and yield of transgenic wheat plants overexpressing vacuolar Na+/H+ antiporter genes
Miroshnichenko D, Poroshin G, Dolgov S
‘Biotron’, Branch of Institute of Bioorganic Chemistry RAS, Pushchino, Moscow Region, Russia
Soil salinity is one of the environmental abiotic stress factors limiting agricultural productivity in many region of the world. Soil salinity severely affects the productivity of cereals, including wheat. One possible mechanism by which plants could survive salt stress is to compartmentalize sodium ions away from cytosol. To improve the plant growth and yield of wheat in saline soils, we have generated transgenic wheat plants overexpressing vacuolar Na+/H+ antiporter genes using the biolistic-mediated transformation method. More than 50 independent transgenic lines with expression of hvnhx2 gene isolated from barley (Hordeum vulgare L.) and agnhx gene isolated from salt-brush (Atriplex gmelini) were produced. Second generation of homozygous transgenic wheat plants showed increased tolerance to salinity (100-200 mM NaCl). Several transgenic wheat plants with higher levels of vacuolar Na+/H+ antiporter transcripts exhibited better biomass production at the vegetative growth stage in saline condition These results demonstrate the feasibility of engineering salt tolerance in plant.
2-35.
The flowering pathway under short day in barley
Kikuchi R1, Kawahigashi H1, Ando T2, Tonooka T3, Handa H1
1Plant Genome Research Unit, National Institute of Agrobiological Sciences, Japan
2Institute of the Society for Techno-innovation of Agriculture, Forestry and Fisheries, Japan
3Barley Research Subteam, National Institute of Crop Science, Japan
Flowering is the essential events for reproductive success in higher plants, and it is significantly affected by photoperiod. Plants classified into three types, long day (LD), short day (SD) or day natural (DN) plants, according to photoperiod response. Barley (Hordeum vulgare) is a facultative LD plant, and its flowering is not only promoted under LD conditions but can also occur even under SD conditions, although it is delayed. FLOWERING LOCUS T (FT), which encodes a protein with a PEBP domain, plays a central role in flowering of Arabidopsis, and the FT homologues of many species have been isolated and analysed. In this study, five barley PEBP genes, HvFT1-3, HvTFL1 and HvMFT1, were analysed to clarify their functional roles in flowering. HvFT1 was expressed at phase transition, and its transgenic rice showed most robust flowering initiation, suggesting that HvFT1 is the key gene for flowering in barley. HvFT2 transgenic rice also showed early heading, but it was expressed only under SD conditions in barley. These results indicate that the role of HvFT2 is limited to SD conditions, unlike HvFT1. HvFT3 was mapped to chromosome 1HL, the same chromosome of Ppd-H2, a major QTL for flowering under SD conditions. HvFT3 was expressed in Morex carrying Ppd-H2, but not in Steptoe carrying ppd-H2. Gene structural analysis revealed that Morex has an intact HvFT3, whereas most of this gene has been lost in Steptoe. These data strongly suggest that HvFT3 is candidate for Ppd-H2, and barley has an adaptive mechanism to adjust flowering even under unfavorable SD conditions using a combination of different FT-like genes.
2-36.
The potential of Hordeum chilense cytoplasm in the development of CMS systems in Triticeae crops
Martín AC
Departamento de Mejora Genética Vegetal, Instituto de Agricultura Sostenible (C.S.I.C.), Apdo. 4084, E-14080 Córdoba, Spain
Cytoplasmic male sterility (CMS) is a maternally inherited trait resulting from incompatibility between nuclear and cytoplasmic genomes characterized by an inability to produce viable pollen but without effects on female fertility. CMS has been generated in several crop species by substituting their normal cytoplasm with the one from closely-related species, while restoration of fertility is generally produced by the introgression of nuclear genes from the cytoplasm donor species. One of the limitations of this system is the deleterious side-effects from the alien cytoplasm as kernel shrivelling, preharvest sprouting and a large reduction in seed germinability amongst others, together with male sterility instability. Hordeum chilense, a diploid wild barley native to Chile and Argentina which posses some traits potentially useful for wheat breeding and which exhibits high crossability with others members of the Triticeae tribe, has been used as cytoplasms donor to produce alloplasmic lines in several crops: Triticum aestivum, Triticum durum, Secale cereale and Triticale. In all of them, alloplasmic lines display the positive characteristics of stable sterility under different environmental conditions and no important side-effects of the cytoplasm as developmental or floral abnormalities, showing only slightly reduced height and some delay in heading. Restorer of pollen fertility appear to be located at least in two H. chilense chromosomes, 1Hch and 6Hch. Considering the features displayed by this cytoplasm, it offers a real potential for the development of viable technology for hybrid cereal production.
2-37.
Agronomic traits and genetic determination of winter wheat lines (Triticum aestivum L.) with multirow spike
Martinek P1, Dobrovolskaya O2,3, Röder MS2, Börner A2
1Agrotest Fyto, Ltd., Havlíčkova 2787, 767 01 Kroměříž, Czech Republic
2Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, 06466, Gatersleben, Germany
3Institute of Cytology and Genetics, SB RAS, Lavrentieva ave 10, 630090 Novosibirsk, Russia
Two groups of spike morphology in wheat (Triticum aestivum L) are distinguished based on the number of spikelets rising from individual nodes of the spike rachis: 1) normal spikelets (NS), where one spikelet rises from one spike rachis node and 2) supernumerary spikelets (SS), where more than one fertile spikelet rises from one rachis node. Attention is given to newly developed winter wheat lines with multirow spike (MRS) which is characterised by a higher number of spikelets with a few florets that rise from individual nodes of spike rachis. The yields of five different lines with MRS were compared with current standard varieties (Akteur, Eurofit, Simila and Rapsodia) with NS on three different levels of nitrogen doses gradated by 30 kg.ha-1 from 90 to 150 kg.ha-1 and at the same level of fungicide treatment; the previous crop was oilseed rape. Nitrogen was applied in the form of limestone ammonium nitrate or urea at the three basic timings: regeneration, production and late topdressings. Each variant of treatment was three times replicated in 10-m2 plots. The average yield of MRS lines in 2007 and 2008 was 96% at low, 99% at medium and 101% at high intensity. The good yield reaction of lines with MRS to nitrogen dose could be explained by higher spike sink capacity of MRS, where higher number of kernels per spike can be developed under higher growing intensities without disease infection. The MRS is determined by one recessive gene which was designated mrs1. The MRS trait was mapped by genotyping F2 populations using microsatellite markers. It was found that Mrs1 locus is located on the chromosome 2DS, about 10cM from the centromere. It will be necessary to improve the genetic background of the MRS lines by hybridisation with important present wheat varieties.
2-38.
Breeding Triticale (X Triticosecale Wittmack) for improved breadmaking quality
Martinek P
Agrotest Fyto, Ltd., Havlíčkova 2787, 767 01 Kroměříž, Czech Republic
Outcomes of breeding triticale (X Triticosecale Wittmack) for the improvement of breadmaking characteristics of grain for making food products from yeast-leavened dough are presented. The breeding programme is based on the employment of triticale donors with translocations of chromosome 1R, to which the segments of wheat chromosome 1D were transferred. These translocations carry the glutenin allele Glu-D1d encoding HMW subunits 5+10. This allele positively affects breadmaking quality. The donors of chromosome 1R translocations were provided by A. J. Lukaszewski (Univ. of California, USA). Two-year data on grain quality (baking and rheological tests) in selected triticale lines from 2007 and 2008 harvests are given. Most grain quality parameters in these lines are close to those of winter wheat classified into B category (bread quality). Though the breadmaking quality of triticale has been partly improved, there has still been a problem of higher α-amylase activity that causes a low falling number, which is usually markedly lower than in wheat. We suppose that the triticale developed for baking purposes will retain most properties typical for this crop: better amino acid composition of storage protein in grain, better ability to use nutrients from soil, generally higher level of health status as compared to wheat, resistance to some unfavourable abiotic factors (particularly tolerance to aluminium ions) and ability to produce good yields in regions where wheat growing is not profitable. It will lead toward extending the use of triticale in the field in which wheat has been dominating until now. The perspective of newly developed translocations in practical breeding is discussed.
2-39.
Gene flow from genetically modified to cultivated wheat plants
Miroshnichenko D, Poroshin G, Dolgov S
‘Biotron’, Branch of Institute of Bioorganic Chemistry RAS, Pushchino, Moscow Region, Russia
One of the most discussed environmental effects associated with the use of transgenic plants is the flow of genes to plants in the environment. Potential risks of gene escape from transgenic crops through pollen and seed dispersal have slowed down full utilization of gene technology in crop improvement. Although no transgenic wheat varieties have yet been officially approved for extensive commercial cultivation in the world, it is apparent that, as an important world cereal crop, transgenic wheat varieties could be released into the environment for commercial production, and probably within the near future.
In 2004-2008 crop-to-crop gene flow in spring wheat was investigated. As the pollen source, a transgenic homozygous line expressing recombinant DNA encoding two marker genes, such as bar and gfp genes was used. Among the marker genes available, the bar gene, encoding phosphinothricin acetyl transferase and conferring resistance to herbicide ammonium glufosinate, are particularly suitable for investigating gene flow in controlled experimental field trials. Green fluorescent protein marker gene gfp was used to monitor in vivo foreign gene expression at different stages of experiments and for segregation study. Analyses of phenotypic and molecular data showed that gene flow was greatly affected by the direction of the dominant wind and the distance between the targets. The overall hybridization rate of transgenic seeds collected from non-transgenic receptor plants growing at one meter from transgenic plants varied year after year from 0.15% (2004) to 0.41% (2005). When non-transgenic receptor plants were grown in plots at two or three meter from transgenic plants, only a few seeds were produced from fertilization with transgenic donor (less than 15% from total amount of transgenic hybrids). A strong asymmetric distribution of the gene flow was detected in different parts of plots and the highest values (0.90%) were recorded following the direction of the dominant wind.
2-40.
Observation of pollen tube growth and molecular mapping of Kr genes in common wheat-rye hybridization
Mishina K1, Manickavelu A2, Sato H1, Katsumata M1, Sassa H1, Koba T1
1Graduate School of Horticulture, Chiba U., 648 Matsudo, Chiba 271-8510, Japan
2Laboratory of Plant Genome Science, Kihara Institute for Biological Research, Yokohama City U., Maioka 641-12, Totsuka-ku, Yokohama 244-0813, Japan
Crossability of common wheat with alien species, e. g., rye, wild and cultivated barley, is known to be controlled by Kr gene family. Reproduction barrier caused by Kr genes decrease hybrid seed set, but molecular mechanisms are still unclear. In this study, we attempted to clarify the pollen tube behaviour of rye on the wheat stigma by using confocal microscope, and to localize the QTLs controlling the crossability in wheat-rye crosses by using molecular markers. In high crossability cultivar Chinese Spring (CS), pollen tubes of rye reached near the micropyle at one hour after pollination, while in low crossability cultivar Hope, Mara and Cheyenne (Cnn), pollen tube growth was inhibited, and pollen tubes seldom reached to the upper part of ovary. These results show that reproduction barrier caused by Kr inhibits alien pollen tube growth. Also we conducted QTL mapping using F2 population consisted of 130 individuals derived from a cross between CS and the chromosome substitution line of CS having chromosome 5B of Cnn, with 58 microsatellite makers. We found two QTL regions controlling the crossability with rye on the chromosome arms of 5BS and 5BL, between the markers WMC47 and BARC142, and between BARC140 and GWM554, respectively. The effect of QTL region on 5BS on crossability with rye was greater than that of 5BL. This result coincident with the report by Tixier et al. (1998), who localized Kr1 and SKr genes on the long and short arms of chromosome 5B, respectively. We are now going to test F6 RIL population derived from the same cross with 91 molecular makers to identify the precise locations of Kr1 and SKr genes on chromosome 5B.
2-41.
Posttranscriptional silencing of CKX genes, regulating cytokinin level in barley by RNA interference
Nadolska-Orczyk A1, Zalewski W1, Galuszka P2, Orczyk W1
1Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland
2Departament of Biochemistry, Palacky University, Olomouc, Czech Republic
Cytokinins are important plant hormones, which play an essential role in plant growth and development. Their level in different tissues is regulated by cytokinin oxidase/dehydrogenase enzymes. The enzymes are coded by a family of CKX genes. Two of them, HvCKX1 and HvCKX2 were isolated in barley (Galuszka et al. 2004). We studied the effect of HvCKX silencing in plants transformed with hpRNAi binary vector pMCG161 (http://www.chromdb.org/mcg161.html). Two vectors containing the fragments of HvCKX1 and the conservative fragments of HvCKX2 in sens and antisens orientation were constructed. The silencing cassettes were introduced into two barley cultivars: Scarlett and Golden Promise by Agrobacterium-mediated (pMCG/CKX1 silencing cassette) and biolistic (pMCG/CKX2 silencing fragment) transformation. The numbers of selected, putative transgenic lines obtained were: 52 for Golden Promise, 1 for Scarlett with pMCG/CKX1 silencing cassette and 5 for Golden Promise and 3 for Scarlett with pMCG/CKX2 silencing cassette. Relative activity of CKX enzyme, measured in roots of T1 seedlings (three repeats of bulk samples) from T0 plants transformed with pMCG/CKX1 silencing cassette ranged from 0.38 to 1.23 of the activity of the wild in vitro plants. Forty of 52 lines of Golden Promise showed significantly lower enzyme activity. There was a positive correlation between enzyme activity and T0 plant productivity (the number of seeds per plant and the mass of thousand kernels) as well as the mass of the roots. Lower CKX activity influenced higher plant productivity and bigger mass of the roots. The relative enzyme activity measured in the roots of the lines transformed with the pMCG/CKX2 silencing cassette was similar to control and ranged from 0.88 to 1.21 in all lines but one - this line showed a significantly higher enzyme activity (2.37 +- 0.02). The productivity of these T0 plants and the root mass from their T1 seedlings were lower comparing with the control. Semiquantitative and quantitative analysis of expression of both genes subjected to silencing - HvCKX1 and HvCKX2 in different tissues of Golden Promise and Scarlett plants has been done. Obtained data were the basis for choosing the most appropriate tissues to study expression/silencing of HvCKX in all transgenic lines.
This research is supported by Ministry of Science and Higher Education; Research Project nr CZECHY/259/2006
2-42.
Genetic variability of MRP gene constituting ‘Qfhs.kibr-2DS’ QTL to reduce Fusarium mycotoxin accumulation among hexaploid wheats
Niwa S1, Kikuchi R2, Handa H2, Ban T1
1Kihara Inst. Biol. Res., Yokohama City U.,Yokohama, Kanagawa 244-0813, Japan
2NIAS, Tsukuba, Ibaraki 305-8602, Japan
The QTL ‘Qfhs.kibr-2DS’, which controls Fusarium mycotoxin accumuration in wheat grains has been reported on 2DS chromosome of hexaploid wheat cv. Gamenya and Nobeokabouzu-komugi. The gene for multi drug resistance-associated protein (MRP) was tracked down as a candidate gene constituting Qfhs.kibr-2DS (Handa et al. 2008). In this report, we demonstrated genetic variability of the MRP gene structure among various hexaploid wheat germplasms. The MRP genes consist of 11 exons on wheat homoeologous chromosome 2A, 2B and 2D were sequenced fully or partially from the BAC library of hexaploid wheat cv. Chinese Spring (CS). To examine genetic variability among wheat germplasms with different level of mycotoxin accumulation, eight sets of genome specific primers for chromosome 2D to amplify several genomic regions of the MRP gene on chromosome 2DS were designed based on the sequence of introns, which connected to exsons. Two types of the MRP gene structure were detected; Chinese wheat type (ex. Sumai 3 and CS) and the others (ex. Gamenya and Nobeokabouzu-komugi) with SNPs and in/del polymorphism. One SNPs marker for the second exon region was developed to identify the MRP genotype for various wheat germplasms collected from the world showing various resistance level to Fusarium head blight, and we will discuss association between MRP genotype and phenotype response on mycotoxin accumulation. This work was supported by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Genomics for Agricultural Innovation, TRC-1005).
2-43.
Alien glutenin subunits expressed in common wheat endosperm affect on the composition
Tanaka H, Arakawa T, Tsujimoto H
Laboratory of Plant Genetics and Breeding Science, Faculty of Agriculture, Tottori University, Japan
The seed storage proteins of common wheat (Triticum aestivum L.) are responsible for the ability of flour to form cohesive dough, required to make strong dough such as bread. However, the narrow genetic base of hexaploid wheat has limited the allelic combinations that are available for the improvement of bread-making quality. Screening for good-quality subunits in wheat related wild species of wheat is important for improving the bread-making quality of wheat. Changes of glutenin composition of wheat endosperm were investigated in 6 alien chromosome addition lines. Each line has group-1 chromosome from wheat related wild species expected to carry glutenin genes. SDS-PAGE and two-dimensional gel electrophoresis have resolved a total of 15 protein bands and 39 protein spots from alien chromosomes, respectively. Agropyron elongatum disomic addition line (DAL) had 11 protein spots with the greatest numbers among them. We also found changes of the spot density from common wheat in DALs. One spot changed to higher density in Aegilops umbellulata DAL, whereas two spots changed to lower density in Hordeum chilense DAL. These observations might show post-translational control and/or changes of gene expression due to presence of additional alien chromosome in common wheat.
2-44.
Characterization of a β-glucanless mutant in barley
Tonooka T1,2, Aoki E1, Yoshioka T1, Taketa S3, Kiribuchi-Otobe C1,2
1National Institute of Crop Science, Tsukuba, Ibaraki 305-8518, Japan
2Graduate school of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan
3Research Institute for Bioresoueces, Okayama University, Kurashiki, Okayama 710-0046, Japan
We reported a novel mutant gene for β-glucanless grain (bgl) in barley (Breeding Science 59:47-54, 2009). In this study, we analysed the effects of the bgl gene on growing properties and quality characteristics using near-isogenic line (NIL). We developed a NIL with bgl using a Japanese two-rowed cultivar ‘Nishinohoshi’ as a recurrent parent. The NIL grew normally in the field and showed normal seed fertility, but had shorter culms and awns than the recurrent parent. The NIL had significantly longer spikes than the recurrent parent; however, the number of spikes was much reduced. Low yield in the NIL was probably caused by the reduced number of spikes. The NIL showed partial chlorosis in the leaves and awns, probably due to a pleiotropic effect of the bgl gene. Heading date was 9 days later in the NIL than in the recurrent parent. β-glucan content in vegetative organs was much reduced in the NIL. The reduction of culm and awn lengths in the NIL might be attributed to the reduction of β-glucan in the cell walls. The content of arabinoxylan in the grains of the NIL was significantly higher than the recurrent parent. The NIL showed completely floury texture in the endosperm. Microscopic observation revealed that the NIL had thinner cell walls in the endosperm. The NIL had soft and friable grain texture, probably due to the thin endosperm cell walls.
2-45.
Virus-induced gene silencing of P23k in barley leaf reveals morphological changes involved in secondary wall formation
Kidou S1, Yokota S2, Yoshida K3
1Cryobiosystem Research Center, Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan
2Faculty of Pharmaceutical Science, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan
3Taisei Corporation Technology Center, Nase344-1 Totuska Yokohama, 245-0051, Japan
P23k is a monocot-unique protein that is highly expressed in the scutellum of germinating barley seed. Previous expression analyses suggested that P23k is involved in sugar translocation and/or sugar metabolism. However, the role of P23k in barley physiology remains unclear. Here, to elucidate its physiological function, BSMV-based virus-induced gene silencing (VIGS) of P23k in barley leaves was performed. Expression and localization analyses of P23k mRNA in barley leaves showed up-regulation of P23k transcript with increased photosynthetic activity and the localization of these transcripts to the vascular bundles and sclerenchyma, where secondary wall formation is most active. VIGS of the P23k gene led to abnormal leaf development, asymmetric orientation of main veins, and cracked leaf edges caused by mechanical weakness. In addition, histochemical analyses indicated that the distribution of P23k in leaves coincides with the distribution of cell wall polysaccharides. Considering these results together, it is proposed that P23k is involved in the synthesis of cell wall polysaccharides and contributes to secondary wall formation in barley leaves. This work was supported partly by the Genomic Agricultural Innovation Program of MAFF (TRC1006).
2-46.
Spontaneous amphidiploidization via unreduced gametes is a universal phenomenon for Triticum turgidum - Aegilops tauschii hybrids
Zhang LQ1,2, Liu DC1,2,3, Zheng YL1,2, Yen Y4
1Triticeae Research Institute, Sichuan Agricultural University, Dujiangyan city, 611830, China
2Key Laboratory of Crop Genetic Resources and Improvement, Ministry of Education, Sichuan Agricultural University, Yaan 625014, China
3Northwest Plateau Institute of Biology, Chinese Academy of Science, Qinghai 810001, China
4Department of Biology and Microbiology, South Dakota State University, Brookings, SD 57007, USA
Polyploidy has been found to be very common in plants. Polyploidy can be formed via the duplication of genomes, either of the same genomes (autopolyploid) or of diverged genomes with homoeologous relationships (allopolyploid). The formation and union of unreduced gametes in triploid F1 hybrid between Triticum turgidum L. (2n=4x=28, AABB) and Aegilops tauschii Coss. (2n=2x=14, DD) might have played an important role in the spontaneous origin of hexaploid wheat (T. aestivum L., 2n=6x=42, AABBDD). Experimental studies have indicated that unreduced gametes by meiotic restitution lead to the fertility of F1 hybrids of T. turgidum and Ae. tauschii and then resulted in spontaneous production of hexaploid wheat. However, previous studies on unreduced gametes were only involved a few T. turgidum genotypes or their combinations with Ae. tauschii. In the present study, triploid F1 plants from 115 wide hybridization combinations were obtained by crossing 76 T. turgidum lines with 24 Ae. tauschii accessions without embryo rescue. It was indicated that a very high frequency (88.70%) of combinations was partial fertile and spontaneously produced hexaploid wheats. The wide spread of spontaneous amphidiploidization in the T. turgidum - Ae. tauschii F1 hybrids found in our study supported the multi-origin hypothesis for the hexaploid wheat by some wheat scientists.
2-47.
Population structure of central Eurasian wild wheat progenitor Aegilops tauschii Coss.
Mizuno N1, Yamasaki M2, Matsuoka Y3, Kawahara T4, Takumi S1
1Graduate School of Agricultural Science, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan
2Food Resources Education and Research Center, Kobe University, Kasai, Hyogo 675-2103, Japan
3Fukui Prefectural University, Matsuoka, Eiheiji, Yoshida, Fukui 910-1195, Japan
4Graduate School of Agriculture, Kyoto University, Muko 617-0001, Japan
Aegilops tauschii Coss. (syn Ae. squarrosa L.) is well known as the D genome progenitor of hexaploid bread wheat. Ae. tauschii, a wild diploid wheat species, has a wide natural species range in central Eurasia, spreading from Turkey to western China. AFLP analysis using total 122 accessions of Ae. tauschii was conducted to clarify the population structure of this wide spreading wild wheat species. Phylogenetic analysis revealed that there were two major lineages, L1 and L2, in the Ae. tauschii population. Bayesian population structure analysis based on the AFLP data showed that L1 and L2 were significantly divided into six and three subgroups, respectively. Only four out of the six L1 subgroups were diverged from western habitats, Transcaucasus and Azerbaijan-Iran Caspian region, to eastern habitats such as Pakistan and Afghanistan. Other subgroups including L2 were limitedly distributed to the western region. Three major haplogroups (HG7, HG9 and HG16) were identified in the Ae. tauschii population based on the chloroplast genome diversity (Matsuoka et al. PLoS ONE 3: e3138, 2008). HG7 accessions were widely distributed to both L1 and L2. HG9 accessions were restricted only to L2, whereas HG16 accessions belonged to L1, suggesting that HG9 and HG16 were formed from HG7 after the two-lineage differentiation of the nuclear genome.
2-48.
Molecular markers for systematic characterization of low molecular weight glutenin subunits in common wheat (Triticum aestivum L.)
XF Zhang1,2, DC Liu1, WL Yang1, JZ Sun1, DW Wang1, HQ Ling1 and AM Zhang1
1State Key Lab of Plant cell and Chromosome engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Chaoyang District, Beijing 100101, China
2Graduate School of the Chinese Academy of Sciences, Beijing 100049, China
Wheat is the staple food for human beings. Wheat flour can be processed into bread, other baked goods (cakes, cookies, crackers etc.), pasta and noodles, and a wide range of other products. Low-molecular-weight glutenin subunits (LMW-GS) are among the major polymeric protein components of wheat flour. Their ability to form intermolecular disulphide bonds with each other and/or with high-molecular-weight glutenin subunits (HMW-GS), is crucial for the formation of glutenin polymers, which determine the technological properties of wheat flour (Weegels et al., 1996; Gupta and Shepherd, 1988). LMW-GS is encoded by a multigene family, displaying high polymorphic protein complexes. Recently, some locus-specific primers of LMW-GS genes have been developed (Wang., 2009; Ikeda et al., 2006; Kawaura et al., 2005; Long et al., 2005; Van Campenhout et al., 1995). Systematic molecular markers specific for each LMW-GS allele would have importance and practical applications in wheat breeding. In this study, we developed systematic molecular markers, referring to the LMW-GS genes isolated from Xiaoyan 54, Glenlea and Norin61 and the sequences published in Genbank. The specificity of these markers, validated in a large collection of wheat varieties from China, was discussed.
2-49.
Genetic diversity of high molecular weight glutenine subunits in wheat landraces
Nishinaka M1, Okumoto Y1, Kato K2, Kawahara T3, Tanisaka T1
1Graduate school of Agriculture, Kyoto University, Kyoto 606-8502, Japan
2Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan
3Graduate school of Agriculture, Kyoto University, Muko 617-0001, Japan
Landraces are useful genetic resources for wheat (Triticum aestivum L.) improvement. However, their potentials have not yet been fully investigated. High-molecular-weight glutenine subunits (HMW-GS) encoded by Glu-A1, Glu-B1, and Glu-D1 loci are major components of wheat seed storage protein, and are greatly related to rheological and bread-making properties. In this study, we aimed to disclose the geographical distributions of HMW-GS alleles using 360 wheat landraces collected from Asia, Europe, Russia, and Africa. HMW-GS composition in each landrace was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) based on the method by Payne and Lawrence (1983). Consequently, three, ten and 13 alleles were identified for Glu-A1, Glu-B1, Glu-D1, respectively. In all the regions, the major genotype for HMW-GS was “null (Glu-A1), 7+8 (Glu-B1), 2+12 (Glu-D1)”. The ratios of these three alleles, however, differed among regions. For Glu-A1 locus, the frequency of 2* allele was larger than 1 allele in the eastern side of the Caspian Sea, South Asia, and Russia, but opposite relationship was found in other regions. Glu-B1 locus exhibited great geographical variations in frequencies of 7+9 and 17+18: the frequencies of these two alleles in South Asia and Russia were quite different from those in their respective neighboring regions. The allelic variation of Glu-D1 locus was quite large in the west coast of the Caspian Sea, especially in Azerbaijan, although extremely small in Iraq, Afghanistan and Iran. A novel allele at Glu-D1 locus was identified from one of the Azerbaijan landraces. To clarify the distribution of this novel allele, additional 297 landraces originated in the regions near the Caspian Sea were investigated, but we found no landraces with this allele.