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Culture efficiency,
represented as the number of green plants per inoculated embryo, was
generally lower in culture No.1 than in the other cultures
(Table
2). Embryos
harvested from intact ear showed less regeneration capacity, that is,
1.2 and 0.4 plants from one inoculated embryo in culture No.2 and
No.3, respectively. On the other hand, more than four plants were
obtained in culture No.2 by culturing embryos pre-treated with 2,4-D
in the later stage of detached ear culture. Similarly more than three
plants were obtained in culture No.3 by culturing the above-mentioned
embryos and the control embryos harvested from detached ear. It was
concluded that regeneration capacity of mature embryo derived callus
decreased by the addition of BAP into subculture medium and increased
by the pre-treatment with 2,4-D in the later stage of detached ear
culture. Though the number of regenerated plants was still low, one
original callus was usually divided into 4 pieces and the maximum
number of regenerated green plants was 16 from one callus piece,
indicationg that more than 60 plants could be obtained from one
inoculated embryo.
In the second experiment, the effect of 2,4-D concentration in
subculture medium on plant regeneration from mature embryo which was
normally harvested from field grown 'IL 68'. Culture procedure was
the same as in culture No.3 (Table
1), except that
1mg/l or 2mg/l of 2,4-D was added to subculture medium. Though no
green spot was formed by subculturing at 2mg/l of 2,4-D even after
100 days of culture, it was formed at 1mg/l of 2,4-D. The number of
calli formed green spot was 0.87 per inoculated embryo. And more than
two plants were obtained from on inoculated embryo. This result
indicated that 2,4-D concentration in subculture medium was also the
important factor for efficient plant regeneration from normal mature
embryo.
Refereneces
Kato K, Tomo S, Yamazaki s and Hayashi K (1990) Simplified
culture method of detached cars and its application to vernalization
in wheat. Euphytica 49: 161-168.
Sasakuma T and Tei A (1989) Plant regeneration and its
characteristics from mature embryo derived callus in common wheat.
Japan J Breed 38 (Suppl. 2); 186-187 (in Japanese).
Shimada T and Yamada Y (1979) Wheat plants regenerated from embryo
cell cultures. Japan J Genet 54: 379-385.
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