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KMID : 0903519900330040351
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1990 Volume.33 No. 4 p.351 ~ p.352
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Soybean tissue culture and transformation
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Kim, Joo Hag
LamotteC E/Hack, E
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Abstract
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A simple and reproducible protocol for regenerating soybean plants from explants derived from 7day-old seedlings has been developed. Explants, each consisting of the primary leaf node(the unifoliate leaf node), the cotyledonary node, the internode between them, and one cotyledon, were cultured in vitro on a modified Murashinge and Skoog(MS) medium containing 3% sucrose, four times the MS inorganic micronutrient level, the vitamins of B5 medium, 2g · 1^(-1) L-proline, 2㎎·1^(-1) N^6-benzyladenine (BA), and 0.02㎎·1^(-1) α-naphthalene acetic acid (NAA) to induce the formation of adventitious shoots at the primary leaf node. In inducing shoot formation, this medium was superior to MS and B5 basal media even when all were supplemented with the same BA and NAA levels. Explants from seedlings of the culLivar Peking produced an average of ca. 30 shoots per explant after 4∼5 weeks on this medium. Shoot formation was greater when a cotyledon remained by medium composition. explant orientation, age of donor seedling, and cultivar. Exogenous cytokinin(BA) was essential for shoot formation : proline and the fourfold-raised level of MS inorganic micronutrients caused a large increase in shoot number.
Adding 2g·1^(-1) L-proline and raising fourfold the level of inorganic micronutrients in the medium enhance soybean shoot regeneration. Proline increased shoot number but decreased length of regenerated shoots, whereas raising micronutrient level increased both shoot number and length and, thus, partly overcame the effect of proline on length of shoots.
Examining separately the effects of singly omiiting, including, raising, and lowering the levels of the seven different micronurtrient elements of the original MS medium has provided evidence that zinc is the most important element for regeneration from explants and that the other six elements, too, are all in less than optimal supply.
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