白菜雄性不育相关基因BcMF3功能的反义RNA验证

为了研究白菜等十字花科植物花粉发育及雄性不育发生的机理,根据编码果胶甲酯酶的白菜雄性不育相关基因BcMF3的cDNA序列设计特异引物,从普通白菜(Brassica campestris ssp.chinensis var.communis,syn.B.rapa ssp.chinensis var.communis)花蕾cDNA中扩增出458 bp的片断,构建BcMF3的反义RNA植物表达栽体,然后转化菜心(B. campestris ssp.chinensis var.parachinensis).研究发现,31.3%菜心转基因植株花粉表现出畸形,而且只有部分花粉具有活力,花粉离体萌发率降低为31.6%,其花药果胶甲酯酶活性降低了12.8%.上述结果表明BcMF3基因与普通白菜和菜心等白菜植物的花粉发育密切相关.

Functional confirmation of BcMF3 gene related to male sterility of Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis) by antisense RNA

Based on the cDNA sequence of BcMF3 gene,which is related to male sterility of Chinese cabbage and encodes a pectin methylesterase(PME),specific primers were designed,and a 458 bp fragment was amplified from the cDNA of flower buds of common Chinese cabbage-pak-choi(Brassica campestris ssp.chinensis var.communis,syn.B.rapa ssp.chinensis var.communis).An expression vector was then constructed using the produced fragment.The Agrobacterium tumefaciens harboring BcMF3 fragment was transformed to flowering Chinese cabbage(B.campestris ssp.chinensis var.parachinensis) via tissue culture.31.3% tested pollen grains from transgenic plants exhibited abnormal shape;Only a portion of the pollen grains from transgenic plants were of vigor,and 31.6% of the pollen grains from transgenic plants germinated normally;The PME activity of the anther from the plants with abnormal pollens was decreased by 12.8%.These results suggested that the product of BcMF3 gene tightly correlates to the pollen development of Chinese cabbage crops such as Chinese cabbage-pak-choi and flowering Chinese cabbage.