本文主要研究内容
作者袁硕硕（2019）在《桑树响应植原体侵染的长链非编码RNA的鉴定及功能研究》一文中研究指出：桑树黄化型萎缩病是一种严重的桑树病害,严重制约了桑树经济和生态价值的实现,目前该病发生的分子机制仍不清楚。本研究采用链特异性转录组测序技术,分析了健康桑树叶片和受植原体侵染(感病)叶片中mRNA和lncRNA的表达谱变化,鉴定出了桑树叶片中响应植原体侵染的mRNAs和lncRNAs,并利用生物信息学技术对其代谢通路进行了分析;在此基础上对其中一个植原体响应基因LRR类受体丝氨酸/苏氨酸蛋白激酶基因(putative LRR receptor-like serine/threonine-protein kinase)(Mul-LRR)和其反义lncRNA(Mul-LRR-AS)的生物学功能及其相互作用进行了研究。本研究的主要结果如下:(1)桑树叶片链特异性转录组测序文库的构建及测序分析成功地构建了桑树健康叶片和感病叶片链特异性转录组测序文库,并利用Illumina Solexa测序技术对其进行了测序分析,共鉴定到99 933个mRNAs和16 805个lncRNAs。在健康叶片和感病叶片差异表达的mRNAs有9 600个,其中在感病叶片中上调的基因有3 049个,下调的基因有6 551个。差异表达mRNAs涉及到生长发育、代谢、信号传导、胁迫响应等多种生理过程。鉴定到的差异表达lncRNAs有943个,其中在感病叶片中上调的有220个,下调的有723个;在差异表达的lncRNAs和mRNAs中有318对被预测存在反义相互作用,其中与抗病相关的有49对。(2)桑树Mul-LRR基因及其反义lncRNA(Mul-LRR-AS)的功能研究利用PCR技术克隆得到了Mul-LRR基因,构建了其植物表达载体,并侵染拟南芥获得了转基因植株。Mul-LRR基因在拟南芥中超表达可以增强转基因植株对丁香假单胞菌番茄致病变种(Pseudomonas syringae pv.Tomato DC3000,Pst.DC3000)的抗性;同时利用PCR技术克隆得到了Mul-LRR-AS,构建了其植物表达载体,并通过侵染拟南芥得到了转基因植株。将转Mul-LRR基因植株与转Mul-LRR-AS基因植株杂交,获得了杂交植株,通过荧光定量PCR分析发现,Mul-LRR-AS在杂交植株成功表达,且Mul-LRR-AS的表达可以抑制Mul-LRR基因的表达。杂交植株与转Mul-LRR基因植株相比对Pst.DC3000的抗性明显降低。表明Mul-LRR-AS可以通过抑制Mul-LRR基因的表达,负调控植株对Pst.DC3000的抗性。(3)桑树Mul-LRR及Mul-LRR-AS基因启动子的表达活性分析利用PCR技术克隆得到Mul-LRR及Mul-LRR-AS基因的启动子序列,分别命名为pMul-LRR和pMul-LRR-AS。两个启动子核苷酸序列中均含有多个CAAT-Box和TATA启动子核心元件,但含有不同的激素和环境响应元件;利用烟草瞬时侵染表达系统结合GUS组织化学染色证明pMul-LRR具有病原菌、水杨酸和茉莉酮酸诱导表达活性,而pMul-LRR-AS不受病原菌、水杨酸和茉莉酮酸处理的诱导。本研究为深入探讨Mul-LRR及其反义lncRNA的生物功能奠定了基础,对于拓宽和丰富植物lncRNA功能和作用机制的认识,全面揭示桑树黄化型萎缩病发生的分子机制具有重要意义,同时也为桑树的抗性育种提供了有效的候选基因。
Abstract
sang shu huang hua xing wei su bing shi yi chong yan chong de sang shu bing hai ,yan chong zhi yao le sang shu jing ji he sheng tai jia zhi de shi xian ,mu qian gai bing fa sheng de fen zi ji zhi reng bu qing chu 。ben yan jiu cai yong lian te yi xing zhuai lu zu ce xu ji shu ,fen xi le jian kang sang shu xie pian he shou zhi yuan ti qin ran (gan bing )xie pian zhong mRNAhe lncRNAde biao da pu bian hua ,jian ding chu le sang shu xie pian zhong xiang ying zhi yuan ti qin ran de mRNAshe lncRNAs,bing li yong sheng wu xin xi xue ji shu dui ji dai xie tong lu jin hang le fen xi ;zai ci ji chu shang dui ji zhong yi ge zhi yuan ti xiang ying ji yin LRRlei shou ti si an suan /su an suan dan bai ji mei ji yin (putative LRR receptor-like serine/threonine-protein kinase)(Mul-LRR)he ji fan yi lncRNA(Mul-LRR-AS)de sheng wu xue gong neng ji ji xiang hu zuo yong jin hang le yan jiu 。ben yan jiu de zhu yao jie guo ru xia :(1)sang shu xie pian lian te yi xing zhuai lu zu ce xu wen ku de gou jian ji ce xu fen xi cheng gong de gou jian le sang shu jian kang xie pian he gan bing xie pian lian te yi xing zhuai lu zu ce xu wen ku ,bing li yong Illumina Solexace xu ji shu dui ji jin hang le ce xu fen xi ,gong jian ding dao 99 933ge mRNAshe 16 805ge lncRNAs。zai jian kang xie pian he gan bing xie pian cha yi biao da de mRNAsyou 9 600ge ,ji zhong zai gan bing xie pian zhong shang diao de ji yin you 3 049ge ,xia diao de ji yin you 6 551ge 。cha yi biao da mRNAsshe ji dao sheng chang fa yo 、dai xie 、xin hao chuan dao 、xie pai xiang ying deng duo chong sheng li guo cheng 。jian ding dao de cha yi biao da lncRNAsyou 943ge ,ji zhong zai gan bing xie pian zhong shang diao de you 220ge ,xia diao de you 723ge ;zai cha yi biao da de lncRNAshe mRNAszhong you 318dui bei yu ce cun zai fan yi xiang hu zuo yong ,ji zhong yu kang bing xiang guan de you 49dui 。(2)sang shu Mul-LRRji yin ji ji fan yi lncRNA(Mul-LRR-AS)de gong neng yan jiu li yong PCRji shu ke long de dao le Mul-LRRji yin ,gou jian le ji zhi wu biao da zai ti ,bing qin ran ni na gai huo de le zhuai ji yin zhi zhu 。Mul-LRRji yin zai ni na gai zhong chao biao da ke yi zeng jiang zhuai ji yin zhi zhu dui ding xiang jia chan bao jun fan jia zhi bing bian chong (Pseudomonas syringae pv.Tomato DC3000,Pst.DC3000)de kang xing ;tong shi li yong PCRji shu ke long de dao le Mul-LRR-AS,gou jian le ji zhi wu biao da zai ti ,bing tong guo qin ran ni na gai de dao le zhuai ji yin zhi zhu 。jiang zhuai Mul-LRRji yin zhi zhu yu zhuai Mul-LRR-ASji yin zhi zhu za jiao ,huo de le za jiao zhi zhu ,tong guo ying guang ding liang PCRfen xi fa xian ,Mul-LRR-ASzai za jiao zhi zhu cheng gong biao da ,ju Mul-LRR-ASde biao da ke yi yi zhi Mul-LRRji yin de biao da 。za jiao zhi zhu yu zhuai Mul-LRRji yin zhi zhu xiang bi dui Pst.DC3000de kang xing ming xian jiang di 。biao ming Mul-LRR-ASke yi tong guo yi zhi Mul-LRRji yin de biao da ,fu diao kong zhi zhu dui Pst.DC3000de kang xing 。(3)sang shu Mul-LRRji Mul-LRR-ASji yin qi dong zi de biao da huo xing fen xi li yong PCRji shu ke long de dao Mul-LRRji Mul-LRR-ASji yin de qi dong zi xu lie ,fen bie ming ming wei pMul-LRRhe pMul-LRR-AS。liang ge qi dong zi he gan suan xu lie zhong jun han you duo ge CAAT-Boxhe TATAqi dong zi he xin yuan jian ,dan han you bu tong de ji su he huan jing xiang ying yuan jian ;li yong yan cao shun shi qin ran biao da ji tong jie ge GUSzu zhi hua xue ran se zheng ming pMul-LRRju you bing yuan jun 、shui yang suan he mo li tong suan you dao biao da huo xing ,er pMul-LRR-ASbu shou bing yuan jun 、shui yang suan he mo li tong suan chu li de you dao 。ben yan jiu wei shen ru tan tao Mul-LRRji ji fan yi lncRNAde sheng wu gong neng dian ding le ji chu ,dui yu ta kuan he feng fu zhi wu lncRNAgong neng he zuo yong ji zhi de ren shi ,quan mian jie shi sang shu huang hua xing wei su bing fa sheng de fen zi ji zhi ju you chong yao yi yi ,tong shi ye wei sang shu de kang xing yo chong di gong le you xiao de hou shua ji yin 。
论文参考文献
论文详细介绍
论文作者分别是来自山东农业大学的袁硕硕，发表于刊物山东农业大学2019-07-08论文，是一篇关于桑树论文,植原体论文,类受体丝氨酸论文,苏氨酸蛋白激酶论文,发病机制论文,山东农业大学2019-07-08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，资料来自山东农业大学2019-07-08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：桑树论文; 植原体论文; 类受体丝氨酸论文; 苏氨酸蛋白激酶论文; 发病机制论文; 山东农业大学2019-07-08论文;