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Materials and methods
Plant materials
Fourteen alloplasmic hybrids used in this study (Table
1) were
originally produced by Prof. K. Tsunewaki (presently Fukui
Prefectural University). CS served as control. Seeds were imbibed for
5hr under tap water, placed in 4C overnight and germinated on filter
papers in petri dishes supplied with either 0 (normal condition) or
0.15 M NaCl solution (salinity condition).
Respiratory measurements
Three-day-old whole seedlings (3 days after imbibition) were used in
the following study of respiration. The rate of oxygen consumption
was measured polarographically using a Klark-type oxygen electrode
(Rank Brothers, Cambridge) according to Nakamura et al. (1991).
Briefly, the whole seedlings were incubated at 25C in 3 nil of buffer
(pH 6.6) containing 50 mM N-[2-hydroxyethyl]
piperazine-N'-2-ethanesulfonic acid and 10 mM
2-[N-morpholino]- ethanesulfonic acid inside the sample cell.
Effects of respiratory inhibitors, i.e., 1 mM KCN for the
cytochrome path and 1 mM salicylhydroxamic acid (SHAM) for the
alternative path, were studied singly and in combination. The data
were taken from at least 9 samples per line and the activity of each
path was calculated according to a model equation developed by Bahr
and Bonner (1973). The residual rate of oxygen consumption which is
resistant to both KCN and SHAM was subtracted to estimate the total
path activity. Average taxonomic distances among all the alloplasmic
hybrids and CS were calculated based on standardized values and a
dendrogram was constructed by UPGMA (Sneath and Sokal 1973).
Results and discussion
The total respiratory path activity at the 3-day-old seedling
stage was significantly higher in 6 alloplasmic hybrids and lower in
3 others than CS under the normal condition (Table
1). D plasm type
of Ae. squarrosa and Mh of Ae. heldreichii
showed higher cytochrome path activities than CS, while
the cytochrome path activity of Sv plasm type of Ae.
kotschyi was lower. Only Cu plasm type of Ae. umbellulata
showed a higher alternative path activity than CS. Under
the normal condition, as a whole, five plasm types (C of Ae.
caudata, Sl of Ae. sharonensis, Sb of Ae.
bicornis and Mt and Mt2 of Ae. mutica) showed the
respiratory path activities equivalent to those of CS.
Under the salinity
condition, coefficients of variation of a majority of parameters were
larger than those under the normal condition (Table
1). Cu plasm type
showed significantly lower total path and cytochrome path activities
than CS, and C and Sv plasm types showed lower total path activities.
It was noted that the alternative path activities of six plasm types
were higher than CS under the salinity condition. The alternative
path is known to be operative uniquely in plants under the conditions
of energy oversupply or limited cytochrome path capacity (Moore and
Siedow 1991). The path is resistant to cyanide and the electron flow
through this path is considered to be energetically wasteful because
of the uncoupling with ATP production. Although physiological
significance of this path remains unclear, it has been suggested that
the path plays a role in maintaining the homeostasis of plant cells
in relation to cellular energy states (Lambers 1982). The observed
higher alternative path activity in one half of the alloplasmic
hybrids than CS under the salinity condition suggests that the energy
overflow through this path is associated with the limited capacity of
the cytochrome path in these alloplasmic hybrids. The response of
plasm types to NaCl salinity, i.e., increases or decreases in the
respiratory path activities relative to those under the normal
condition, is shown in Fig.
1. NaCl salinity
caused significant increases in the total and cytochrome path
activities in CS. The observation is consistent with the report that
the cytochrome path activity increases in wheat plants subjected to
NaCl salinity, possibly to cope with the increased energy demand
under salinity conditions (Zagdanska 1995). similarly, Sl, Mt, Mt2
and Sv plasm types showed increases in both total and cytochrome path
activities under the salinity condition. On the other hand, the
salinity condition caused significant decreases in the total path
activity and/or cytochrome path activity in D, Mh and G plasm
types.
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