白菜花药特异基因BcPRO的克隆及表达分析

Isolation and expression analysis of anther-specific gene BcPRO from Brassica campestris ssp.chinensis(L.) Makino.

根据基因数据库已有的Profilin 4序列设计简并引物,在白菜花蕾cDNA中成功克隆到前纤维蛋白基因,命名为BcPRO,在基因库中的登陆号为EF492988.BcPRO基因编码区由405个核苷酸组成,编码134个氨基酸,推导的氨基酸序列与甘蓝型油菜和拟南芥的花粉前纤维蛋白分别有95%和92%的同源性.RT-PCR表明,BcPRO基因仅在保持系的Ⅲ级、Ⅳ级和Ⅴ级花蕾及Polima雄性不育系的Ⅲ级和Ⅳ级花蕾中表达,并且表达量逐级增大,而在莲座叶、花茎叶、茎、根及Ogura雄性不育系的所有部位均未能检测到.进一步研究发现,BcPRO基因在花药中表达,在萼片、花丝、雌蕊、花瓣等部位均未见表达.这些实验结果表明,BcPRO基因为花药特异基因,可能在花粉发育过程中起着重要作用,并且在相同核背景下,有着不同的细胞核-细胞质互作模式.文中还就不同植物花粉前纤维蛋白氨基酸序列进行了比对、分析.

A gene, designated BcPRO, was isolated from flower buds using gene-speeifie primer pairs and was submitted to GenBank under accession number EF492988. The complete coding DNA sequence of BcPRO was 405 bp in length encoding 134 amino acids and shared more than 900//oo identity with those of Brassica napus and Arabidopsis thaliana. RT PCR showed that BcPRO gene was expressed in stage Ⅲ , IV and V of flower buds of maintainer and in stage Ⅲ and IV of flower buds of Polima cytoplasmic male sterility （CMS）. However, there was no detection in all tissues of Ogura CMS. RT-PCR revealed BcPRO gene was expressed only in anthers, not in sepals, petals, filaments and pistils. These results suggested that BcPRO gene was an anther-specific one and might play important roles in pollen development. Though under the same nuclear background, BcPRO gene showed different nucleocytoplasmic interaction patterns. Additionally, comparisons and analysis of different pollen profilin amino acids sequences were performed,