摘要
目的:克隆大理天门冬Asparagus taliensis膨胀素(Expansin)相关基因及其启动子,并分析其序列特征与表达模式。方法:基于前期大理天门冬转录组测序数据,利用RT-PCR克隆了一个根系中优势表达的基因AtaEXPB2,并进行生物信息学分析。利用实时荧光定量PCR法(qRT-PCR)检测AtaEXPB2基因的表达模式。结果:AtaEXPB2基因是编码一个含有256个氨基酸残基的膜表面蛋白的全长为771 bp的序列。启动子序列特征分析发现,启动子区域除了含有保守元件外,还存在大量响应元件,这些响应元件与植物体的非生物胁迫和激素处理响应相关。对AtaEXPB2基因的特异性表达分析表明,该基因的表达量在贮藏根和吸收根地下部分显著高于地上部分。qRT-PCR分析表明,逆境胁迫及激素处理会诱导AtaEXPB2基因的表达,逆境胁迫包括干旱胁迫和冷胁迫;激素处理包括脱落酸(ABA)和茉莉酸甲酯(MeJA)。结论:从大理天门冬中成功克隆了AtaEXPB2基因及其启动子,该基因表达量在贮藏根和吸收根中较高,并受冷和干旱胁迫以及ABA、MeJA激素的诱导,推测该基因可能参与大理天门冬逆境胁迫应答。
Objective:To clone the expansin related genes and promoters of Asparagus taliensis,and analyze their sequence characteristics and expression patterns.Methods:Based on the transcriptome sequencing data of Asparagus taliensis,a gene AtaEXPB2 with dominant expression in the root system was cloned using RT-PCR and subjected to bioinformatics analysis.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression pattern of AtaEXPB2 gene.Results:AtaEXPB2 gene was a full-length sequence of 771 bp encoding a membrane surface protein containing 256 amino acid residues.The analysis of promoter sequence characteristics showed that the promoter region contained not only conservative elements,but also a large number of response elements,which were related to abiotic stress and hormone treatment response of plants.The specific expression analysis of AtaEXPB2 gene showed that the expression of ATAEXPB2 gene was significantly higher in underground parts such as storage roots and absorption roots than above ground parts.qRT-PCR analysis showed that the expression of AtaEXPB2 gene was induced by adverse stress and hormone treatment,the adverse stress included drought stress and cold stress,the hormone treatments included abscisic acid(ABA)and methyl jasmonate(MeJA).Conclusion:The AtaEXPB2 gene and its promoter are successfully cloned from Asparagus taliensis.The expression of ATaEXPB2 gene is higher in storage roots and absorption roots,and it is induced by cold and drought stress and ABA and MeJA hormones,suggesting that this gene may be involved in the response to adverse stress of Asparagus taliensis.
作者
赖雨馨
张淑惠
李云宾
林春
毛自朝
刘正杰
LAI Yu-xin;ZHANG Shu-hui;LI Yun-bin;LIN Chun;MAO Zi-chao;LIU Zheng-jie(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;The Key Laboratory for Crop Production and Smart Agriculture of Yunnan Province,Kunming 650201,China)
出处
《中药材》
北大核心
2025年第5期1074-1081,共8页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金项目(32360089)
云南省基础研究计划面上项目(202201AT070254)。
