紫花苜蓿MsJMT基因的克隆及其与育性的关联分析  

Cloning of MsJMTGene from Alfalfa and its Correlation with Fertility

作　　者：徐小博 王鑫尧 郁伟杰 苗一凡 徐博[1] XU Xiao-bo;WANG Xin-yao;YU Wei-jie;MIAO Yi-fan;XU Bo(College of Forestry and Pratacultural Science,Jilin Agricultural University,Changchun,Jilin Province 130118,China)

机构地区：[1]吉林农业大学林学与草学学院,吉林长春130118

出　　处：《草地学报》2022年第9期2306-2314,共9页Acta Agrestia Sinica

基　　金：吉林省科学技术厅科技发展计划项目(20210202069nc);吉林省教育厅科学技术研究项目(JJKH20210371KJ);吉林农业大学国家级大学生创新创业训练计划项目(CXXM022)资助。

摘　　要：以实验室前期对紫花苜蓿(Medicago sativa L.)不育系BSA-Seq分析为背景,为探究茉莉酸类对其育性的影响,本试验采用PCR法,成功从紫花苜蓿基因组中克隆了一个全长1047bp,功能为控制茉莉酸O-甲基转移酶合成的基因,命名为MsJMT。分析表达模式后发现,该基因编码氨基酸348个,编码蛋白为不稳定非分泌亲水蛋白。亚细胞定位结果为核质表达。构建基因进化树后,验证其氨基酸序列与5种豆科植物相似度达99%以上。利用qPCR技术分析其对紫花苜蓿花苞育性的时空表达差异,结果显示不育系全时期表达量比保持系高且在Ⅲ时期两者差异极显著(P<0.01)。而茉莉酸含量测定结果则相反,为不育系全时期低于保持系,且保持系Ⅲ时期含量最高。利用外源茉莉酸甲酯喷施处理Ⅲ时期花苞,结果为在0.5mmol·L^(-1),1mmol·L^(-1)水平下花药提前开裂。研究结果表明MsJMT可能通过茉莉酸通路影响紫花苜蓿育性。To explore the effects of Jasmonates(JAs)on the fertility of alfalfa(Medicago sativa L.)under the background of per-laboratory BSA-Seq analysis on sterile lines,this study successfully cloned a 1047 bp gene from alfalfa genome by PCR method.The function is to control jasmonate O-methyl transferase synthesis gene,named MsJMT.Analysis of the expression pattern showed that the gene encoded 348amino acids,and the encoding protein was an unstable non-secretory hydrophilic protein.Subcellular localization results showed nuclear and cytoplasmic expression.The amino acid sequence of the gene was 99%similar to that of five leguminous plants.qPCR was used to analyze the spatio-temporal expression differences in the bud fertility of alfalfa.The results showed that the expression levels of sterile lines were upregulated and significantly higher than those of maintainers in stageⅢat the 0.01level.However,the Jasmonate(JA)content of flower buds in stagesⅠtoⅣwas opposite.The results showed contrasted that the JA content in the sterile line was lower than that in the maintainer line during the whole period,and the highest was in the maintainer lineⅢ.The buds of stageⅢwere treated by spraying with exogenous Methyl jasmonate(MeJA).The results showed that the anther degenerated in advance at 0.5mmol·L^(-1) and 1 mmol·L^(-1)levels.The conclusion was that MsJMT may affect alfalfa fertility through the JA pathway.

关 键 词：茉莉酸甲基转移酶 紫花苜蓿 基因克隆 序列分析 不育系 

分 类 号：S963.223.3[农业科学—水产养殖]