通过花粉管通道法导入红树总DNA获得耐盐紫花苜蓿T0代植株及其RAPD验证

利用花粉管通道法将盐生植物红树总DNA导入紫花苜蓿,共导入1 391朵花,获得894粒T₀代转化种子。T₀代种子种植在含有225 mmol/L NaCl的MS培养基上,获得12株耐盐性强的植株。以供体和受体为对照,对12株耐盐植株进行RAPD分析,在55条随机引物中有8条扩增出稳定、清晰的条带,条带的差异性表现为新增条带、供体特异条带和受体条带丢失。以上结果初步证实外源DNA已经整合到受体的基因组中,而且T₀代植株耐盐能力提高可能与外源基因的导入有关。

Production and RAPD verification of T0 salt-tolerant alfalfa introducing the total DNA from Rhizophora apiculata by the pollen-tube pathway

The exogenous total DNA of Rhizophora apiculata was introduced into alfalfa (Medicago sativa) by the pollen-tube pathway. In this study, 1 391 flowers were treated and 894 transformed T₀ seeds were harvested. Twelve T₀ seedlings with improved salt-tolerance were obtained when the T₀ seeds were planted in MS medium containing 225 mmol/L NaCl. RAPD analysis of T₀ seedlings, receptor and donor were carried out. Eight primers which could amplify steady and clear bands were chosen from 55 arbitrary primers tested, and increased band diversity with new bands, specific bands in donor and deletion in receptor. It showed that the exogenous DNA was conformed for integration into the receptors genome, and the improved salt-resistance of T₀ seedlings may be related to the introduction of exogenous genes.