| 摘要: |
| 【目的】克隆分析紫花苜蓿MsTIFY11基因的分子特征，并探讨干旱胁迫、茉莉酸（JA）和盐胁迫对其表达水平的影响，为进一步研究MsTIFY11基因调控苜蓿抗旱性和耐盐性提供参考。【方法】培育草原4号紫花苜蓿幼苗，在45 d时分为对照组及干旱、JA和盐胁迫组，对照组定时补水，在结荚期采集根、茎、老叶、嫩叶、花、荚和种子样品，用于MsTIFY11组织表达特异性分析；干旱胁迫组不浇水，于处理0，2，4，8，12和24 h采集叶片样品；JA胁迫组在植株上喷洒100 μmol/L 茉莉酸甲酯液，在处理0，1，3，6 和12 h采集叶片样品；盐胁迫处理组浇灌250 mmol/L NaCl液，在处理0，2，4，8，12 和24 h采集叶片样品。PCR克隆MsTIFY11基因，采用生物信息学方法分析基因结构、蛋白质性质以及亚细胞定位。利用RT-qPCR检测不同组织及干旱、盐、JA胁迫下MsTIFY11基因的表达量。【结果】MsTIFY11基因编码区长721 bp，编码240个氨基酸残基，含有TIFY和CCT_2结构域。氨基酸序列分析表明，MsTIFY11蛋白为不稳定蛋白;同源序列比对和系统发育分析发现,紫花苜蓿与蒺藜苜蓿（Medicago truncatula）、红三叶（Trifolium pratense）、鹰嘴豆（Cicer arietinum）、豌豆（Pisum sativum）、白车轴草（Trifolium repens）和地三叶（Trifolium subterraneum）的亲缘关系较近。亚细胞定位预测结果显示，MsTIFY11位于细胞核。RT-qPCR分析发现,MsTIFY11基因在各个组织中均有表达，其中在根的表达量最高；在干旱和JA胁迫下苜蓿叶片中MsTIFY11的表达量显著升高，在盐胁迫下其表达量总体呈上升趋势。【结论】MsTIFY11基因的表达受干旱、盐和茉莉酸诱导。 |
| 关键词: 紫花苜蓿 MsTIFY11基因 组织表达模式 非生物胁迫 |
| DOI：10.13207/j.cnki.jnwafu.2025.11.007 |
| 分类号: |
| 基金项目:内蒙古自治区高等学校青年科技英才支持计划项目（NJYT23009）；苜蓿分子育种体系构建及种质创制项目（BR22-11-12）；内蒙古农业大学草学学科青年基金项目（IMAUCXQJ2023004） |
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| Cloning and expression analysis of MsTIFY11 in Medicago sativa L. |
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ZHANG Yan1, CHEN Qi1, DAI Rui1, HUO Xiaowei1, GUO Na1, ZHANG Zhiqiang2
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1.College of Grassland Science,Inner Mongolia Agricultural University,Hohhot，Inner Mongolia 010000,China;2.College of Grassland Science,Technology Engineering Center of Drought and Cold-resistant Grass Breeding in North of the National Forestry and Grassland Administration,Inner Mongolia Agricultural University,Hohhot，Inner Mongolia 010000,China
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| Abstract: |
| 【Objective】This research aimed to conduct cloning analysis on the molecular characteristics of the alfalfa（Medicago sativa L.）MsTIFY11 gene,and explore the effects of drought stress,jasmonic acid (JA) and salt stress on its expression level,to provide a reference for further research on the regulation of drought resistance and salt tolerance of alfalfa by the MsTIFY11 gene.【Method】Medicago sativa.L.cv.‘Caoyuan No.4’ seedlings were cultivated and divided into control group,drought stress group,JA group and salt stress group at day 45.Seedlings in the control group was regularly replenished with water,and their roots,stems,old leaves,young leaves,flowers,pods and seeds were collected during the pod-setting stage for tissue-specific analysis.Leaf samples of the seedlings in the drought stress group were collected at 0,2,4,8,12 and 24 h.Methyl jasmonate solution of 100 μmol/L was sprayed on the seedlings in the JA group，leaf samples were collected at 0,1,3,6 and 12 h.250 mmol/L NaCl solution was applied to the seedlings in the salt stress group,and leaf samples were collected at 0,2,4,8,12 and 24 h.The MsTIFY11 gene was cloned by PCR,and bioinformatics methods were used to analyze the gene structure,protein properties,and subcellular localization prediction.RT-qPCR was used to detect the expression level of MsTIFY11 gene in different tissues under drought,salt,and JA stress.【Result】The results showed that the coding region of MsTIFY11 gene is 721 bp long,encoding 240 amino acid residues,and contains TIFY and CCT_2 domains.Amino acid sequence analysis showed that MsTIFY11 protein is an unstable protein.Phylogenetic analysis showed that MsTIFY11 protein is close to Medicago truncatula,Trifolium pratense,Cicer arietinum,Pisum sativum,Trifolium repens and Trifolium subterraneum.Subcellular localization prediction showed that MsTIFY11 is located in the nucleus.RT-qPCR analysis showed that the MsTIFY11 gene was expressed in various tissues, with the highest expression level observed in roots.The MsTIFY11 expression level in alfalfa leaves increased significantly under drought and JA stress,and the MsTIFY11 expression level generally increased under salt stress.【Conclusion】The expression of MsTIFY11 gene is induced by drought,salt and jasmonate stress. |
| Key words: Medicago sativa L. MsTIFY11 gene tissue expression pattern abiotic stress |
