生物技术通报2025，Vol.41Issue(8)：234-241,8.DOI:10.13560/j.cnki.biotech.bull.1985.2025-0059
红花CtbHLH128基因克隆及调控干旱胁迫应答功能研究
Cloning of Gene CtbHLH128 in Safflower and Response Function Regulating Drought Stress
摘要
Abstract
[Objective]Basic helix-loop-helix(bHLH)transcription factors represent one of the largest families of transcription factors in eukaryotes,and their members play crucial roles in plant growth,development,and stress responses.This study enriches the research on the function of the bHLH transcription factor family genes and lays an important foundation for analyzing the molecular regulatory mechanism of safflower(Carthamus tinctorius L.)seed development.[Method]Based on the complete CDS sequence of CtbHLH128 in the genome,primers were designed using Primer 6.0 software to clone and obtain the correct sequence of the gene,and bioinformatics was used to analyze the number of amino acids encoded by the gene,conserved domains,and phylogenetic tree.The CtbHLH128 fragment was constructed into a plant expression vector and transformed into Arabidopsis thaliana by Agrobacterium infection.Herbicide screening and PCR were used to identify positive strains.Drought stress was applied to the positive strains at the seedling and germination stages to explore the role of this gene in drought stress.[Result]The CtbHLH128 consists of 948 bp nucleotides,encoding 315 amino acids.The molecular weight of its encoded protein is 33 742.32 Da,and the theoretical isoelectric point is 9.02.CD-Search analysis results revealed that its C-terminus contained a typical HLH conserved domain located at amino acid residues 249-301.Phylogenetic tree revealed high similarity with bHLH128 gene in sunflower and lettuce.The CtbHLH128 gene was overexpressed in Arabidopsis,and three T3 positive strains were obtained through resistance screening and PCR identification.The positive strains were subjected to drought stress-related phenotypic analysis.The results revealed that transgenic Arabidopsis plants showed greater sensitivity to drought conditions at the seedling stage,but enhanced resistance to drought at the germination stage.[Conclusion]This suggests that the CtbHLH128 gene may possess potential functions in the responses to drought stress and seed germination.关键词
红花/转录因子/CtbHLH128/基因克隆/序列分析/过表达/干旱胁迫Key words
safflower/transcription factor/CtbHLH128/gene cloning/sequence analysis/overexpression/drought stress引用本文复制引用
李开杰,吴瑶,李丹丹..红花CtbHLH128基因克隆及调控干旱胁迫应答功能研究[J].生物技术通报,2025,41(8):234-241,8.基金项目
贵州省科技计划项目(黔科合基-ZK[2022]一般064) （黔科合基-ZK[2022]一般064）
