Boyce Thompson Institute, Cornell University, 14853, Ithaca, NY, USA.
Plant Cell Rep. 1985 Dec;4(6):344-7. doi: 10.1007/BF00269895.
A tissue culture procedure for the regeneration of somatic embryos and plantlets from somatic cells of the soybean Glycine max is described. Bean pods of soybean cv. TGM119 were immersed in liquid nitrogen for 20 minutes. Young embryos were excised from the immature seeds and cultured to form calli. Calli grown from the young embryos were incubated in liquid culture for two weeks. The liquid suspension culture was filtered to obtain single cells. The soybean cells were cultured for one month in a liquid medium in hanging drop cultures for development into proembryoids. The proembryoids were maintained on a solid growth medium for 40 days. The resultant callus tissue was transferred into MS media containing selected combinations and concentrations of 2,4-Dichlorophenoxyacetic acid, Naphthaleneacetic acid, Kinetin, Benzyladenine and Indoleacetic acid. In the presence of Benzyladenine (0.2 mg/l) and Indoleacetic acid (0.01 mg/l), globular and heart shaped somatic embryos were formed on the surface of the calli. Calli containing somatic embryos were transferred into liquid medium and incubated under low light conditions. After six months further incubation, more than 1,000 plantlets and a large number of somatic embryoids at various developmental stages were obtained per flask.
介绍了从大豆体细胞再生体细胞胚和植株的组织培养程序。将大豆 cv. TGM119 的豆荚在液氮中浸泡 20 分钟。从未成熟的种子中切下幼胚并培养形成愈伤组织。从小胚中生长出来的愈伤组织在液体培养中培养两周。液体悬浮培养物经过过滤以获得单细胞。将大豆细胞在悬滴培养的液体培养基中培养一个月，以发育成原胚。原胚在固体生长培养基上保持 40 天。所得愈伤组织被转移到含有选择的组合和浓度的 2,4-二氯苯氧乙酸、萘乙酸、激动素、苄基腺嘌呤和吲哚乙酸的 MS 培养基中。在苄基腺嘌呤（0.2mg/l）和吲哚乙酸（0.01mg/l）的存在下，在愈伤组织的表面形成球形和心形的体细胞胚。含有体细胞胚的愈伤组织被转移到液体培养基中，并在低光照条件下培养。进一步培养六个月后，每个培养瓶中获得了超过 1000 株和大量处于不同发育阶段的体细胞胚状体。
