结缕草ZjCCS基因的克隆与表达分析

为了解Cu/Zn超氧化物歧化酶分子伴侣基因(CCS)在结缕草逆境胁迫过程中的表达调控机制,以结缕草(Zoysia japonica)盐胁迫条件下的转录组数据为依据,结缕草cDNA为模板,通过RT-PCR法,克隆了924 bp长的结缕草ZjCCS基因的CDS序列。同源性分析显示该基因与谷子(Setaria italica)同源性最高,含有植物CCS蛋白所特有的3个保守结构域。利用实时荧光定量PCR(qRT-PCR)技术研究该基因在非生物胁迫条件下在结缕草叶片中的表达情况,结果显示,在盐胁迫条件下,ZjCCS基因的表达量呈先上升后下降的趋势,4 h表达量最高,相对于盐胁迫,干旱和重金属胁迫对ZjCCS基因的表达量影响不大,推测ZjCCS基因可能在结缕草响应盐胁迫过程中发挥作用。

Cloning and Expression Analysis of ZjCCS from Zoysia Japonica

A chaperone gene (ZjCCS) of copper superoxide dismutase gene of Zoysia japonica was cloned by RT-PCR. The ZjCCS gene contains a 924-nucleotides-long open reading frame(ORF), which encoded a protein of 307 amino acids. Its expression level was analyzed for elucidating its role in response to salt drought and heavy-metal stress. Homology Blast (BlastP) with the deduced amino acids sequence indicated that ZjCCS gene had a high similarity to other plant CCS genes, and the predicted ZjCCS polypeptide contained 3 distinct domains. QRT-PCR experiments showed that ZjCCS mRNA level in leaf was significantly induced by salt stress, and reached to peak after 4 h of salt treatment. Drought stress and heavy-metal stresses didn't have a significant influence on the ZjCCS expression level, which suggested that ZjCCS might play a major role when zoysia japonica responding to salt stress.