摘要
根据苦荞花期转录组数据,采用RT-PCR技术和PCR技术克隆得到一条新的b HLH类蛋白基因FtbHLH3(登录号:KU296217),并分析了逆境胁迫下该基因在芽期苦荞胚轴和子叶中的表达量。结果显示,Ftb HLH3基因cDNA全长1 273 bp,包含一个957 bp的开放阅读框,编码蛋白含318个氨基酸。蛋白结构域分析表明,FtbHLH3编码蛋白N-端和C-端分别含有一个bHLH家族典型的结构域。系统进化树分析表明,FtbHLH3与其它植物参与抗逆的bHLH蛋白聚为一簇。荧光定量PCR分析表明,UV-B处理苦荞后,胚轴中FtbHLH3相对表达量在6 h内缓慢增加至1.08,12 h后显著上升至48 h的2.96;而子叶中FtbHLH3相对表达量在3 h后就显著上升,至24 h达到最大值6.64后趋于稳定。4℃冷处理苦荞后,胚轴和子叶中FtbHLH3相对表达量均随时间持续上升,并在48 h后达到最大值,分别为3.22和10.27。可见,本研究克隆的Ftb HLH3基因可能参与了苦荞对UV-B和寒冷等非生物胁迫的应答反应。
Based on the transcriptome data of Fagopyrum tataricum at flowering, a novel FtbHLH3 （GenBank accession number： KU296217） bHLH-like protein gene was amplified from Fagopyrum tataricum by using RT-PCR and PCR techniques. Meanwhile, FtbHLH3 expression profiles was analyzed when tartary buckwheat at the stage of germination was treated under abiotic stress. The result indicated that FtbHLH3 was 1 273 bp in length, and open reading frame of 957 bp encoding a protein of 318 amino acids residues. Sequence analysis showed that the FtbHLH3 gene owned the characteristics of bHLH transcriptional factor known. Seen from the phylogenetic trees, FtbHLH3 was gathered into the same cluster with other bHLH protein related to the stress resistance. The quantitative real-time PCR （qRT-PCR） analysis indicated the FtbHLH3 expression was increased persistently and obviously after UV-B radiation and 4℃ cold treatment. Accordingly, FtbHLH3 gene would be involved in the abiotic stress response such as UV-B radiation and cold presumably.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第2期429-435,共7页
Genomics and Applied Biology
基金
四川省国际合作项目(2015HH0047)基金资助
关键词
苦荞
bHLH转录因子
基因克隆
非生物胁迫
Tartary buckwheat, bHLH transcription factor, Gene cloning, Abiotic stress
