摘要
目的:克隆红腺忍冬Lonicera hypoglauca Miq.的MYB基因并对其进行生物信息学及其功能分析。方法:以转录组测序获得的MYB基因序列进行引物设计,采用qRT-PCR方法克隆MYB基因全长;通过在线软件及工具对该基因所编码的氨基酸序列进行生物信息学分析。比较不同浓度PEG6000胁迫处理24 h后的植株叶片MYB基因表达模式,并与相对应的叶片含水量、叶绿素含量、超氧化物歧化酶(SOD)活性和可溶性蛋白质含量4个生理指标的变化进行相关分析。结果:成功获得MYB基因编码区全长909 bp,编码一个由302个氨基酸组成的蛋白质;DNA结合区域位于N端,长53个氨基酸残基,形成3个螺旋,其中含有两个间距为19个氨基酸残基的色氨酸;这段区域的第3个螺旋含有MYBR1转录因子CCA1-like/R-R亚族的特征模块SHAQKYF;该基因受到PEG诱导调节,其相对表达量随着PEG处理浓度的升高而增加,并与叶片中的含水量、叶绿素含量、可溶性蛋白质含量和SOD酶活性相关系数分别为-0.6964、-0.9132、-0.3714和0.1483。结论:克隆获得预期的红腺忍冬LhMYB1基因,并确认此基因参与了植株的干旱胁迫生理响应,该研究为今后深入开展红腺忍冬耐旱优良品质的研究奠定了基础。
Objective:To clone the MYB gene from Lonicera hypoglauca and analyze its bioinformatics and function.Methods:The primers were designed based on MYB gene sequences obtained by transcriptome sequencing.The full length of MYB gene was cloned by qRT-PCR.Online software and tools were used for bioinformatics analysis of the amino acid sequence encoded by the gene.The expression patterns of MYB gene in leaves were compared after 24 hours stress treatment with different concentration of PEG6000,and the correlation analysis was carried out with the changes of the corresponding leaf water content,chlorophyll content,superoxide dismutase(SOD)activity and soluble protein content.Results:The coding region of MYB gene was 909 bp,encoding a protein consisting of 302 amino acids.The DNA binding region was located at the N-terminal,which was 53 amino acid residues long,forming three helical,including two tryptophan with 19 amino acid residues spacing.The third helix in this region contained SHAQKYF,the signature module of the MYBR1 transcription factor CCA1-like/R-R subfamily.The gene was regulated by PEG induction,it relative expression of this gene increased with the increase of PEG treatment concentration,and the correlation coefficients with water content,chlorophyll content,soluble protein content and SOD activity in leaves were-0.6964,-0.9132,-0.3714,0.1483,respectively.Conclusion:LhMYB1 gene of Lonicera hypoglauca is cloned and confirmed to be involved in physiological response of plant to drought stress,the study provides basis for further research on drought tolerance quality of Lonicera hypoglauca.
作者
师凤华
谭木秀
刘凤鸣
莫乔程
SHI Feng-hua;TAN Mu-xiu;LIU Feng-ming;MO Qiao-cheng(Guangxi Botanical Garden of Medicinal Plants,Nanning 530023,China)
出处
《中药材》
CAS
北大核心
2022年第7期1577-1581,共5页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金项目(31760422)
广西自然科学青年科学基金项目(2018GXNSFBA281144)
关键词
红腺忍冬
MYB转录因子
生物信息学
干旱胁迫
Lonicera hypoglauca Miq.
MYB transcription factor
PEG
Information biology
Drought stress
作者简介
师凤华(1976-),女,博士后,副研究员,研究方向:药用植物生物技术,Tel:18076333929,E-mail:fenghuas@163.com。
