广西植物2018，Vol.38Issue(2)：202-209,8.DOI:10.11931/guihaia.gxzw201706013
银杏bHLH91转录因子基因的克隆及表达分析
Cloning and expression analysis of a bHLH91 transcription factor gene from Ginkgo biloba
摘要
Abstract
bHLH transcription factor plays an important role in plant growth, stress response and secondary metabolism. In order to study the function of bHLH transcription factor gene in Ginkgo biloba,we isolated a bHLH gene from G. biloba and carried out bioinformatics and expression analyses.In this study,a cDNA sequence of bHLH gene was isolated from G. biloba by PCR, which was designated as GbbHLH91. DNA sequencing and sequence analysis showed that the amplified GbbHLH91 gene was 1 425 bp, GbbHLH91 gene contained a complete open reading frame, and the open reading frame of GbbHLH91 was 1 065 bp,encoding 354 amino acids.The predicted GbbHLH91 protein had a mo?lecular mass of 40.1 kDa with isoelectric point value of 8.20.Homology analysis with BLASTP and Align X indicated that the putative GbbHLH91 share a high identical with other known bHLH proteins from different plant species.Phylogenetic analysis showed that the GbbHLH91 protein was closely related to bHLH protein from Pinus tabuliformis,and was 60%identity to bHLH from Amborella trichopoda, which indicated the bHLH gene was relatively conserved during evolution. Real?time PCR assay found that GbbHLH91 gene was expressed in all tested tissues of Ginkgo biloba, while expression level in leaf was the highest,followed by root and stem,the GbbHLH91 gene was lowly expressed in female flowers and fruits of G.biloba,and the lowest in male flower.The expression level of GbbHLH91 gene in G.biloba leaves at different developmental stages was also different. The expression level of GbbHLH91 gene was the highest in mid?April, and then the expression level of the gene showed a decreasing trend with the growth and development of G.biloba leaves. These results provide a preliminary basis for further validation of the GbbHLH91 gene function.关键词
银杏/bHLH/转录因子/基因克隆/表达分析Key words
Ginkgo biloba/bHLH/transcription factors/gene clone/express analysis分类
农业科技引用本文复制引用
何昌文,朱丽,沈珊,张威威..银杏bHLH91转录因子基因的克隆及表达分析[J].广西植物,2018,38(2):202-209,8.基金项目
长江大学博士基金(801190010127)[Supported by the Doctor Foundation of Yangtze University(801190010127)]. （801190010127）
