霸王的原生质体培养的研究

目的：为利用原生质体融合技术转移霸王抗旱基因。方法：采用酶解法分离霸王原生质体,比较了霸王子叶和愈伤组织游离原生质体的产量和活力,不同渗透压和起始密度对原生质体分裂频率的影响。结果：愈伤组织游离的原生质体产量和活力均高于子叶,原生质体产率可达2.4×106个/g.FW,活力达89%。采用液体浅层培养,在附加2,4-D（2mg/L）、6-BA（1.0mg/L）、2%蔗糖和甘露醇（0.4mol/L）的DPD培养基中,原生质体分裂频率最高,达68.6%。转移到附加2-iP（3mg/L）、KT（1.0mg/L）、6-BA（1.0mg/L）的分化培养基上,获得2个再生苗。结论：采用酶解法游离霸王愈伤组织,可获得高活力和高分裂频率的霸王原生质体。

Studies on Protoplasts Culture of Zygophyllum xanthoxylum

Objective:To transfer resistent gene of Zygophyllum xanthoxylum by protoplast fusion.Method: Z.xanthoxylum protoplasts were obtained through enzyme digestion method,they were compared that the yield and viability of protoplasts of cotyledon and calli from Z.xanthoxylum and protoplast division frequecy in different osmotic potentia and densities.Result: The protoplast yield and viability from calli of Zygophyllum xanthoxylum were higher than those from the cotyledons,respectively 2.4×10~6/g·FW and 89%.The protoplast cell division frequency was up to 68.6% in DPD medium containing 2,4-D(2mg/L),6-BA(1.0mg/L), 2% sucrose and mannitol(0.4mol/L) by liquid thin layer culture.Two regenerated plantlets were obtained when these calli were transferred to MS medium containing 2-iP(3mg/L),KT(1.0mg/L),6-BA(1.0mg/L).Conclusion: the high protoplast iability and cell division frequency of were obtained through enzyme digesting calli of Z.xanthoxylum.