白菜核雄性不育两用系花蕾的mRNA差别显示及其cDNA差异片段分析

Differential display of flower bud mRNA of genic male sterility (GMS) AB line in Chinese cabbage-pak-choi and analysis of differential cDNA fragments

用 m RNA差别显示技术对白菜核雄性不育两用系中不育株群与可育株群花蕾总 RNA进行了比较分析 .在 6种反转录模板和 2 0种差异随机引物共 12 0种组合的选择扩增中共回收了 18个 c DNA差异片段 ,对其中 6个 c DNA差异片段进行 Northern验证 ,获得了一个白菜核雄性不育两用系可育株群并在花蕾中特异表达的阳性差异片段 ,简称为 B0 30 2 - 4 .Northern验证结果表明它在不育株中的表达受到了明显的抑制 ,其长度为 4 6 3bp.经 Gen Bank BLAST查询 ,它与拟南芥第一条染色体上编码细胞色素 P4 5 0的基因 CYP86 C4核苷酸序列有 84 .3%的同源性 .

The difference was compared by mRNA differential display technology in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee) flower bud among genic male sterility (GMS) line (A line) and maintainer line (B line). 18 differential fragments have been obtained in total of 120 selective amplifications. A differential fragment named as B0302-4 was confirmed by Northern hybridization out of 6 cDNA differential fragments tested. B0302-4 was shared by the maintainer line, the expression of B0302-4 was restricted in the GMS line, and B0302-4 expressed specially in flower bud. Fragment B0302-4 is composed of 463 base pair. The result of GenBank BLAST showed that it has 84.3% nucleotide similarity to the CYP86C4 gene encoding cytochrome P450 from Arabidopsis.