REGENERATION OF FERTILE PLANTS FROM PROTOPLASTS OF SOYBEAN (GLYCINE-MAX L MERR) - GENOTYPIC DIFFERENCES IN CULTURE RESPONSE
REGENERATION OF FERTILE PLANTS FROM PROTOPLASTS OF SOYBEAN (GLYCINE-MAX L MERR) - GENOTYPIC DIFFERENCES IN CULTURE RESPONSE
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DOI：
10.1007/bf00235083
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发表时间：
1992-06-01
影响因子：
6.2
通讯作者：
WIDHOLM, JM
中科院分区：
文献类型：
--
作者：
DHIR, SK;DHIR, S;WIDHOLM, JM
14种大豆(Glyine max L.)利用先前描述的克拉克63品种的程序，评估了不同基因型从原生质体再生的能力。从未成熟的子叶组织中分离出原生质体，在含有不同糖的KP8、MS或B5的液体或琼脂糖凝胶中培养。电镀效率差异显著，JACK和A-2396的电镀效率高达63%，而X-3337的电镀效率低至38%。用KB培养基常规稀释后，在5~6周内形成直径1~2 mm的菌落，所有检测的基因型别均为阳性。然后将这些菌落转移到MSB(MS Salts；Murashige&Skoog，1962+B5有机物；Gamborg等人，1968)+2，4-D、BA和KN各0.5mgL-1和CH500mgL-1的培养基上进一步生长。当菌落变绿、致密、结节，大小为8-10 mm时，转移到再生培养基中。在常规继代培养中，A-2396、张伯伦、黑龙江26、杰克、雷斯尼克和XP-3015等6个基因型的愈伤组织均能分化出不定芽，其中杰克的愈伤组织再生率最高，达27%(192个愈伤组织中有52个产生了8-12个不定芽)。不同基因型的再生不定芽伸长生根。到目前为止，已经获得了63台完整的设备，包括12台A-2396，19台张伯伦，15台杰克，9台雷斯尼克和8台XP-3015。总共有35株植物被移植到温室的盆栽中。16个已经结实，其他的正在开花和结荚。
Fourteen soybean (Glycine max L.) genotypes were evaluated for their regenerability from protoplasts using a procedure previously descibed for the cultivar Clark 63. Protoplasts were isolated from immature cotyledon tissue and were cultured in liquid or agarose gelled KP8, MS or B5 medium with different sugars. Significant differences were observed in plating efficiency, which was as high as 63% in Jack and A-2396, and as low as 38% in X-3337. Upon regular dilution with KB medium, 1-2 mm diameter colonies were formed in 5-6 weeks with all the genotypes tested. These colonies were then transferred onto MSB (MS salts; Murashige & Skoog, 1962 + B5 organics; Gamborg et al., 1968) medium with 0.5 mg l-1 each of 2,4-D, BA and KN and 500 mg l-1 CH for further growth. Once the colonies had become green, compact and nodular, and were 8-10 mm in size, they were transferred to regeneration medium. Upon regular subculturing, calli of six genotypes; A-2396, Chamberlain, Heilong-26, Jack, Resnick and XP-3015 developed shoots, with the regeneration frequency being highest 27% in Jack (52 calli out of 192 produced 8-12 shoots). The regenerated shoots from different genotypes were elongated and rooted. So far, sixty three complete plants have been obtained, including twelve A-2396, nineteen Chamberlain, fifteen Jack, nine Resnick and eight XP-3015. A total of thirty five plants have been transplanted into pots in the greenhouse. Sixteen have set seeds and others are producing flowers and pods.