本文主要研究内容
作者王玉洁（2019）在《MtENOD20在蒺藜苜蓿共生固氮过程中的功能研究》一文中研究指出：本试验以蒺藜苜蓿为研究对象,对实验室前期筛选出的Phytocyanins基因家族中的早期结瘤素基因MtENOD20（MTR4g130800）进行组织定位、亚细胞定位、RNA干扰和过量表达等分析,探究其在共生固氮中的功能。研究结果简述如下:1、目的基因的表达模式分析利用qPCR方法分析了早期结瘤素基因MtENOD20的两个可变剪接MtENOD20-1、MtENOD20-2在蒺藜苜蓿共生结瘤过程中的时空表达情况,结果表明（1）MtENOD20-1与MtENOD20-2在接菌的根中均被强烈诱导但表达谱不同,在不接菌对照根中不表达或仅有低水平的表达;（2）MtENDO20-1在不接菌和接菌植物的去瘤根、茎、叶中几乎不表达,只在根瘤中表达,为结瘤特异性表达;MtENDO20-2在接菌植物的去瘤根、茎、叶表达量较低,在根瘤中高水平表达,为结瘤增强性表达。推测MtENOD20的两个可变剪接在根瘤发育中可能行使着不同的功能。2、MtENOD20的组织特异性表达构建MtENOD20启动子与GUS融合表达载体,通过发根农杆菌介导的转化体系,对转化根进行GUS染色观察。结果显示,目的基因主要在侵染根的皮层细胞（即根瘤原基起始的部位）、含有侵染线的根毛细胞以及根瘤侵染区表达。3、MtENOD20的亚细胞定位构建MtENOD20基因的亚细胞定位载体,利用根癌农杆菌介导的烟草叶片细胞瞬时转化体系明确了MtENOD20定位在质膜、内质网和细胞核上;发根农杆菌介导的蒺藜苜蓿根部转化体系试验表明,MtENOD20定位于共生体上。这些结果表明MtENOD20可能通过内质网的分泌途径靶向共生体。4、MtENOD20基因RNA干扰和过量表达转化植株的表型分析对MtENOD20进行干扰处理,结果显示:干扰植株的株高、根长减少,根瘤数目、固氮酶活和豆血红蛋白含量降低,且根瘤多为小的白色无效根瘤;接菌后7d的侵染事件统计结果表明,干扰植株中卷曲根毛、延伸的侵染线、侵染线和根瘤原基的数目均降低与对照相比;根瘤石蜡切片和透射电镜观察结果表明,与野生型相比,干扰根瘤中的类菌体分化程度低,为不具有固氮活性的I类或者II类类菌体,共生关系过早终止。共生相关基因的表达分析显示,与对照相比,干扰根中与侵染相关的基因MtNSP1、MtERN1、MtERN2和MtERN3没有变化,MtNIN、MtENOD11和MtENOD40表达量下降,推测目的基因可能在MtNSP和MtERN下游发挥作用;与类菌体分化相关的基因RSD、DNF2、NCR001和NCR035,在干扰根中均显著降低。以上结果表明,MtENOD20可能与根瘤菌的侵染过程及类菌体的释放、分化有关。过表达分析结果显示,MtENOD20过表达植株的株高、根长、根部的干重和鲜重及叶绿素的含量均增加,根瘤的数目、固氮酶活和豆血红蛋白含量上升;侵染事件统计分析结果表明,接菌后7d,过表达根中根毛卷曲、延伸侵染线、侵染线和根瘤原基的数目与对照相比均增加;接菌后45d根瘤石蜡切片和透射电镜观察结果显示,过量表达根瘤细胞中的类菌体形态规则,排列紧密,对照组根瘤中的类菌体数量减少且出现融合现象,表明对照的根瘤细胞开始衰老。
Abstract
ben shi yan yi ji li mu xu wei yan jiu dui xiang ,dui shi yan shi qian ji shai shua chu de Phytocyaninsji yin jia zu zhong de zao ji jie liu su ji yin MtENOD20（MTR4g130800）jin hang zu zhi ding wei 、ya xi bao ding wei 、RNAgan rao he guo liang biao da deng fen xi ,tan jiu ji zai gong sheng gu dan zhong de gong neng 。yan jiu jie guo jian shu ru xia :1、mu de ji yin de biao da mo shi fen xi li yong qPCRfang fa fen xi le zao ji jie liu su ji yin MtENOD20de liang ge ke bian jian jie MtENOD20-1、MtENOD20-2zai ji li mu xu gong sheng jie liu guo cheng zhong de shi kong biao da qing kuang ,jie guo biao ming （1）MtENOD20-1yu MtENOD20-2zai jie jun de gen zhong jun bei jiang lie you dao dan biao da pu bu tong ,zai bu jie jun dui zhao gen zhong bu biao da huo jin you di shui ping de biao da ;（2）MtENDO20-1zai bu jie jun he jie jun zhi wu de qu liu gen 、jing 、xie zhong ji hu bu biao da ,zhi zai gen liu zhong biao da ,wei jie liu te yi xing biao da ;MtENDO20-2zai jie jun zhi wu de qu liu gen 、jing 、xie biao da liang jiao di ,zai gen liu zhong gao shui ping biao da ,wei jie liu zeng jiang xing biao da 。tui ce MtENOD20de liang ge ke bian jian jie zai gen liu fa yo zhong ke neng hang shi zhao bu tong de gong neng 。2、MtENOD20de zu zhi te yi xing biao da gou jian MtENOD20qi dong zi yu GUSrong ge biao da zai ti ,tong guo fa gen nong gan jun jie dao de zhuai hua ti ji ,dui zhuai hua gen jin hang GUSran se guan cha 。jie guo xian shi ,mu de ji yin zhu yao zai qin ran gen de pi ceng xi bao （ji gen liu yuan ji qi shi de bu wei ）、han you qin ran xian de gen mao xi bao yi ji gen liu qin ran ou biao da 。3、MtENOD20de ya xi bao ding wei gou jian MtENOD20ji yin de ya xi bao ding wei zai ti ,li yong gen ai nong gan jun jie dao de yan cao xie pian xi bao shun shi zhuai hua ti ji ming que le MtENOD20ding wei zai zhi mo 、nei zhi wang he xi bao he shang ;fa gen nong gan jun jie dao de ji li mu xu gen bu zhuai hua ti ji shi yan biao ming ,MtENOD20ding wei yu gong sheng ti shang 。zhe xie jie guo biao ming MtENOD20ke neng tong guo nei zhi wang de fen bi tu jing ba xiang gong sheng ti 。4、MtENOD20ji yin RNAgan rao he guo liang biao da zhuai hua zhi zhu de biao xing fen xi dui MtENOD20jin hang gan rao chu li ,jie guo xian shi :gan rao zhi zhu de zhu gao 、gen chang jian shao ,gen liu shu mu 、gu dan mei huo he dou xie gong dan bai han liang jiang di ,ju gen liu duo wei xiao de bai se mo xiao gen liu ;jie jun hou 7dde qin ran shi jian tong ji jie guo biao ming ,gan rao zhi zhu zhong juan qu gen mao 、yan shen de qin ran xian 、qin ran xian he gen liu yuan ji de shu mu jun jiang di yu dui zhao xiang bi ;gen liu dan la qie pian he tou she dian jing guan cha jie guo biao ming ,yu ye sheng xing xiang bi ,gan rao gen liu zhong de lei jun ti fen hua cheng du di ,wei bu ju you gu dan huo xing de Ilei huo zhe IIlei lei jun ti ,gong sheng guan ji guo zao zhong zhi 。gong sheng xiang guan ji yin de biao da fen xi xian shi ,yu dui zhao xiang bi ,gan rao gen zhong yu qin ran xiang guan de ji yin MtNSP1、MtERN1、MtERN2he MtERN3mei you bian hua ,MtNIN、MtENOD11he MtENOD40biao da liang xia jiang ,tui ce mu de ji yin ke neng zai MtNSPhe MtERNxia you fa hui zuo yong ;yu lei jun ti fen hua xiang guan de ji yin RSD、DNF2、NCR001he NCR035,zai gan rao gen zhong jun xian zhe jiang di 。yi shang jie guo biao ming ,MtENOD20ke neng yu gen liu jun de qin ran guo cheng ji lei jun ti de shi fang 、fen hua you guan 。guo biao da fen xi jie guo xian shi ,MtENOD20guo biao da zhi zhu de zhu gao 、gen chang 、gen bu de gan chong he xian chong ji xie lu su de han liang jun zeng jia ,gen liu de shu mu 、gu dan mei huo he dou xie gong dan bai han liang shang sheng ;qin ran shi jian tong ji fen xi jie guo biao ming ,jie jun hou 7d,guo biao da gen zhong gen mao juan qu 、yan shen qin ran xian 、qin ran xian he gen liu yuan ji de shu mu yu dui zhao xiang bi jun zeng jia ;jie jun hou 45dgen liu dan la qie pian he tou she dian jing guan cha jie guo xian shi ,guo liang biao da gen liu xi bao zhong de lei jun ti xing tai gui ze ,pai lie jin mi ,dui zhao zu gen liu zhong de lei jun ti shu liang jian shao ju chu xian rong ge xian xiang ,biao ming dui zhao de gen liu xi bao kai shi cui lao 。
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论文详细介绍
论文作者分别是来自西北农林科技大学的王玉洁，发表于刊物西北农林科技大学2019-07-11论文，是一篇关于共生固氮论文,干扰论文,基因过表达论文,基因表达分析论文,西北农林科技大学2019-07-11论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自西北农林科技大学2019-07-11论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：共生固氮论文; 干扰论文; 基因过表达论文; 基因表达分析论文; 西北农林科技大学2019-07-11论文;