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Wheat Information Service
Number 76: 70-72 (1993)


Structure analysis of mitochondria genomes in interspecific hybrids related to Aegilops triuncialis

Y. Ogihara, M. Furuya and T. Sasakuma

Kihara Institute for Biological Research, Yokohama City University, Nakamura-cho 2-120-3, Yokohama 232, Japan


Mitochondria genome of angiosperm is known to show some characteristic features such as; 1) plasticity of genome structure, 2) promiscuous DNA, and 3) RNA editing. In order to analyze structural alterations of mitochondria genomes in interspecific hybrids related to Aegilops triuncialis (CCUU as genome formula), Restriction Fragment Length Polymorphism (RFLP) analysis of mitochondrial DNAs in Ae. triuncialis complex was carried out. Ae. triuncialis complex was used for the study, because various combinations of interspecific hybrids (natural hybrids, ancestral species, artificially synthetic strains and alloplasmic lines) were obtained.

Total DNAs were isolated from the young seedlings of 20 strains of Ae. triuncialis complex (
Table1) according to the CTAB method (Murray and Thompson 1980). These DNAs were digested with Bam HI and Hind III, and hybridized with twelve mitochondrial genes as probes (Table 2) to carry out RFLP analysis. RFLP analysis of total chloroplast DNAs from five strains of Ae. triuncialis as well as common wheat was also conducted.

Chloroplast DNAs of five strains were classified into three groups, namely C type, U type and the third type (Murai and Tsunewaki 1986), the two of which revealed the identical fragment pattern with those of Ae. caudata (C type) and Ae. umbellulata (U), respectively (Ogihara and Tsunewaki 1988). No ancestral strains corresponding to the third type of Ae. triuncialis was found, so far examined.

Based on the fragment patterns produced from Southern hybridization in combination of twelve mitochondrial gene probes with two restriction enzymes, namely 24 probe-enzyme combination, similarity among eighteen strains was calculated. Using the similarity value, a dendrogram showing genetic relationships among eighteen strains were constructed according to the UPGMA (Nei 1975) as shown in
Fig.1. Mitochondrial DNAs of Ae. triuncialis were classified into three major groups, namely C type, U type and the third type. The mitochondrial DNA of the third type of Ae. triuncialis (#) was much closer to the C type rather than the U type. These lines of evidence support the data obtained from RFLP analysis of chloroplast DNAs. But intraspecific variations were found in both mitochondria DNAs of C and U groups. It is striking that mitochondrial DNA of Jones Fife was diversed from other mitochondrial DNAs of B type (Fig.1), although the fragment patterns of chloroplast DNA of Jones Fife was identical to that of other common wheat. It is noteworthy that mitochondrial genomes of interspecific hybrids were different from those of original strains: Fragment patterns of trn1-trn2, S-trn1, S-trn2, and J02 differed from those of cdt1. Simultaneously, those of trn4-trn7, and trn8 were different from that of umb1. Cdt1 and umb1 were the cytoplasm donors of hybrid species of Ae. triuncialis including synthetic strains and alloplasmic lines. This suggests that mitochondrial DNAs were possibly changed during and/or after interspecific hybridization. These lines of evidence indicate the plasticity of mitochondria genome in wheat complex.


References

Murray HG and Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acid Res 8: 4321-4325.

Murai K and Tsunewaki K (1986) Molecular basis of genetic diversity among cytoplasms of Triticum and Aegilops species.IV CtDNA variation in Ae. triuncialis. Heredity 57: 335-339.

Nei M (1975) Molecular population genetics and evolution. North Holland.

Ogihara Y and Tsunewaki K (1988) Diversity and evolution of chloroplast DNA in Triticum and Aegilops as revealed by restriction fragment analysis. Their Appl Genet 76: 321-332.

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