埃斯基红豆草下胚轴愈伤组织原生质体的培养与植株再生

CULTURE AND PLANT REGENERAION OF PROTOPLASTS FROM HYPOCOTYL CALLI OF SAINFOIN(Onobrychis viciae folia Scop.)CULTIVAR ESKI

埃斯基红豆草幼苗的下胚轴切段在附加２，４－Ｄ０．５ｍｇ／Ｌ，ＫＴ１ｍｇ／Ｌ的ＭＳ中形成胚性愈伤组织。来自１１－１３个月龄、继代６－１５天的愈伤组织的原生质体，在改良的Ｖ－ＫＭ液体培养基中可持续分裂形成细胞团，培养１０天时的分裂率和克隆率分别为６５．８８％和５３．３８％，５周后就可将原生质体形成的小愈伤组织转于固体培养基上。原生质体在改良的Ｂ＿５液体培养基也可以分裂形成小愈伤组织，但分裂率低于Ｖ－ＫＭ。来自原生质体的小愈伤组织在加有０．５％琼脂糖的培养基上可以旺盛生长，并有根、芽的分化，继代子ＭＳ附加ＢＡ０．５－１ｍｇ／Ｌ、ＮＡＡ０．１－０．５ｍｇ／Ｌ的分化基中发育成苗，在附加ＢＡ０．０２ｍｇ／Ｌ、ＮＡＡ０．２ｍｇ／Ｌ的１／２ＭＳ中苗分化出根而再生植株。

Embryogenic calli was able to be formed from sainfoin cv. Eski hypocotyl explants culturedon MS medium containing 2,4-D0.5mg/L,KT1mg/L. Protoplasts were isolated from the callicultured continuously for 11-13 months after being transferred 6-15 days. When cultured inmodified liquid V-KM,the protoplast sustained division and formed cell colonies. At 10 days,thefrequency of cell division and colony formation are 65.88% and 53.38% respectively. Microcallicould be transferred in semisolid media after 5 weeks. The protoplasts were also to divide andforme microcalli in the modified B_5 medium,but the division freqtiency was lower than in V-KMmedium. Protoplasts-derived microcalli exhibited active growth and regenerated roots and buds whentransferred onto solid (0.5%agrose)media. Shoots were obtained from the calli on MS mediumplus BA 0.5-1mg/L and NAA 0.1-0.5mg/L. Roots were induced from the shcots on 1/2MS medium plus BA 0.02mg/L and NAA 0.2mg/L.