胞质雄性不育相关基因的克隆及其表达分析

根据GenBank中orf224和atp6基因的保守序列设计特异引物,对不结球白菜Pol胞质雄性不育系的基因组DNA进行特异扩增,获得了Pol胞质雄性不育的特异基因orf224和atp6的DNA序列.用Blastn在GenBank中进行同源性比对分析,发现不结球白菜orf224c和atp6c基因与已报道的orf224和atp6基因的同源性高达100%,在GenBank中的登录号依次为DQ400846、DQ412559.运用半定量RT-PCR对orf224c基因在不同器官中的表达水平进行分析,结果表明:不育系花期叶片中该基因的表达量比长度小于0.5 mm蕾和雄蕊中的明显低,在后2个器官中的表达量无明显差异;而在保持系的上述3个器官中的表达量无明显差异.该结果显示orf224基因表达上调与孢原细胞分化受阻相关.

Cloning and Expression of Genes Related with Cytoplasmic Male Sterility

Specific primers were designed according to the conserved sequences of orf224 and atp6 genes in GenBank and DNA sequences of orf224 and atp6 genes specific to pol cytoplasmic male sterility were obtained by amplifying genomic DNA of pol cytoplasmic male sterility line of no-heading Chinese cabbage with the primers.By means of Blastn the DNA sequences of orf224c and atp6c gene in no-heading Chinese cabbage were aligned with and found as high as 100% homologous to those of reported orf224 and atp6 genes in GenBank and their accession codes were DQ400846 and DQ412559.The expressions of orf224c gene in different organs were analyzed by semi-QRT-PCR indicating that in the male sterility line the gene had lower expression in the leaves than in the flower buds and anthers that were less than 0.5 mm long at the flowering stage and there were no expression differences between the latter two organs;in the maintainer there were no obvious expressions between the three organs.This result indicated that the up-regulation expression of orf224 gene was related to the suffocation of sporogonium differentiation.