本文主要研究内容
作者徐雅静（2019）在《CAMTA3对植物抗核盘菌及水稻白叶枯病菌的调控功能及机制分析》一文中研究指出：由核盘菌(Sclerotinia sclerotiorum)引起的菌核病和稻黄单胞菌(Xanthomonas oryzae pv.oryzae,Xoo)所致的白叶枯病分别是我国油料和蔬菜作物及水稻的重要病害。抗病调控分子机制的进一步解析有助于为这些病害的防控提出新的策略、提供更好的理论指导。钙信号途径是植物生长发育和胁迫响应的重要调控途径。CAMTA(Calmodulin-binding transcription activator)家族转录因子是结合 Ca2+受体CaM的Ca2+信号途径重要因子。其中CAMTA3对植物基础抗性(Basal resistance)的重要调控作用及机制已经明确,但其对其他类型的抗病性如诱导抗性(Induced resistance)和非寄主抗性(Nonhost resistance)的调控作用及分子机制尚不明确。另外,钙肥作为植物钙补充剂已经在生产上得到广泛应用。然而,钙肥对植物抗病性的诱导作用研究很少,其调控分子机制还不明确。本论文以核盘菌和Xoo为病原物对象,以对核盘菌的基础抗性、诱导抗性以及对Xoo的非寄主抗性为抗性对象,研究钙肥外源施用和CAMTA3对抗病性的调控作用及分子机制。获得的主要结果如下:(1)阐明了 CAMTA3在拟南芥对核盘菌的基础抗性和Pepl激发的诱导抗性以及对Xoo的非寄主抗性中的广谱调控作用,揭示了基于CAMTA3-CBP60g/SA兄Dl模块的水杨酸(Salicylic acid,SA)合成调控新途径及其在上述抗性中的重要作用。拟南芥cbp60g-1和sard1-1单、双突变体以及sid1、sid2/ics1和npr1-1等功能丧失型(Loss-of-function)突变体接种分析结果表明,CBP60g/SARD1-ICS1介导的SA合成和信号传导途径对上述三类抗性起正调控作用。拟南芥功能丧失型突变体camta3及功能获得型突变体(Gain-of-function mutant)camta3-3D接种分析结果表明,CAMTA3对上述三类抗性起负调控作用。拟南芥camta3及camta3-3D突变体中CBP60和SARD1基因表达分别显著上调和下调;凝胶阻滞分析显示CAMTA3 CG-1结构域和CBP60g基因、但不和SA兄D1基因启动子中CGCG box强烈、特异地结合;启动子活性LUC报告分析表明CAMTA3在植物体内负调控CBP60g和SARD1似基因表达,而Pep1和Xoo减弱CAMTA3的这种负调控作用。说明CAMTA3直接负调控CBP60g、可能间接负调控SARD1,而Pep1和Xoo抑制CAMTA3该调控功能。这些结果表明,CAMTA3通过直接负调控CBP60g、可能间接负调控SARD1,抑制ICS1介导的SA合成,从而负调控抗病性;而Pep1和Xoo通过抑制CAMTA3该功能从而诱导抗病性。(2)明确了钙肥外施对核盘菌抗性的诱导作用及调控分子机制。钙肥(有效成分为CaCl2,0.3 mg/ml)叶面喷雾显著诱导拟南芥和油菜产生对核盘菌的抗性,抗性持续期在5天以上。同样钙肥处理不能诱导cbp60g-1和sard1-1突变体产生这种抗性。LUC报告分析显示,钙肥抑制了 CAMTA3对CBP60g和SARD1的负调控作用,但不显著改变CAMTA3基因表达,表明钙肥通过抑制CAMTA3蛋白对CBP60g和SARD1的负调控作用,释放CBP60g和SARD1,从而激发抗病性。(3)鉴定获得了 茉莉酸(Jasmonic acid,JA)、活性氧(Reactive oxygen species,ROS)和RNA干扰(RNA intefering,RNAi)通路基因中的CAMTA3靶标基因,揭示CAMTA3参与调控这些通路。启动子序列结果显示,JA途径基因JAZ1、JAZ2、JAZ5、JAZ7、JAZ8和 MYC2、ROS 通路 NADPH 氧化酶基因RbohF 以及 RNAi通路基因AGO4 AGO5、AGO6、RDR1、RDR3、RDR4的启动子含有 CGCGbox。EMSA分析表明拟南芥CAMTA3 CG-1结构域在体外直接结合AGO4、AGO5、AGO6、RDR3、RBOHF、JAZ1和MYC2的启动子区CGCG box。启动子活性LUC报告分析发现在植物体内拟南芥CAMTA3正调控AGO5和JAZ1的转录表达。综上所述,本研究阐明了拟南芥CAMTA3对核盘菌的基础抗性和诱导抗性以及对Xoo的非寄主抗性的广谱调控功能,揭示了基于CAMTA3以CBP60g和SARD1为靶标的SA合成调控新途径,明确了以该途径为基础的广谱抗病调控新机制。鉴定了 JA、ROS和RNAi通路中的CAMTA3靶标,揭示了 CAMTA3对这些通路的调控作用。明确了钙肥对核盘菌抗性的诱导作用,揭示了钙肥通过抑制CAMTA3对CBP60g和SARD1的负调控作用激发抗病性的分子机制。这些作用和机制的揭示增进了植物抗病分子机制的理解。Pep1和钙肥诱抗作用的发现,还为菌核病防控提供潜在的新方法。
Abstract
you he pan jun (Sclerotinia sclerotiorum)yin qi de jun he bing he dao huang chan bao jun (Xanthomonas oryzae pv.oryzae,Xoo)suo zhi de bai xie ku bing fen bie shi wo guo you liao he shu cai zuo wu ji shui dao de chong yao bing hai 。kang bing diao kong fen zi ji zhi de jin yi bu jie xi you zhu yu wei zhe xie bing hai de fang kong di chu xin de ce lve 、di gong geng hao de li lun zhi dao 。gai xin hao tu jing shi zhi wu sheng chang fa yo he xie pai xiang ying de chong yao diao kong tu jing 。CAMTA(Calmodulin-binding transcription activator)jia zu zhuai lu yin zi shi jie ge Ca2+shou ti CaMde Ca2+xin hao tu jing chong yao yin zi 。ji zhong CAMTA3dui zhi wu ji chu kang xing (Basal resistance)de chong yao diao kong zuo yong ji ji zhi yi jing ming que ,dan ji dui ji ta lei xing de kang bing xing ru you dao kang xing (Induced resistance)he fei ji zhu kang xing (Nonhost resistance)de diao kong zuo yong ji fen zi ji zhi shang bu ming que 。ling wai ,gai fei zuo wei zhi wu gai bu chong ji yi jing zai sheng chan shang de dao an fan ying yong 。ran er ,gai fei dui zhi wu kang bing xing de you dao zuo yong yan jiu hen shao ,ji diao kong fen zi ji zhi hai bu ming que 。ben lun wen yi he pan jun he Xoowei bing yuan wu dui xiang ,yi dui he pan jun de ji chu kang xing 、you dao kang xing yi ji dui Xoode fei ji zhu kang xing wei kang xing dui xiang ,yan jiu gai fei wai yuan shi yong he CAMTA3dui kang bing xing de diao kong zuo yong ji fen zi ji zhi 。huo de de zhu yao jie guo ru xia :(1)chan ming le CAMTA3zai ni na gai dui he pan jun de ji chu kang xing he Peplji fa de you dao kang xing yi ji dui Xoode fei ji zhu kang xing zhong de an pu diao kong zuo yong ,jie shi le ji yu CAMTA3-CBP60g/SAxiong Dlmo kuai de shui yang suan (Salicylic acid,SA)ge cheng diao kong xin tu jing ji ji zai shang shu kang xing zhong de chong yao zuo yong 。ni na gai cbp60g-1he sard1-1chan 、shuang tu bian ti yi ji sid1、sid2/ics1he npr1-1deng gong neng sang shi xing (Loss-of-function)tu bian ti jie chong fen xi jie guo biao ming ,CBP60g/SARD1-ICS1jie dao de SAge cheng he xin hao chuan dao tu jing dui shang shu san lei kang xing qi zheng diao kong zuo yong 。ni na gai gong neng sang shi xing tu bian ti camta3ji gong neng huo de xing tu bian ti (Gain-of-function mutant)camta3-3Djie chong fen xi jie guo biao ming ,CAMTA3dui shang shu san lei kang xing qi fu diao kong zuo yong 。ni na gai camta3ji camta3-3Dtu bian ti zhong CBP60he SARD1ji yin biao da fen bie xian zhe shang diao he xia diao ;ning jiao zu zhi fen xi xian shi CAMTA3 CG-1jie gou yu he CBP60gji yin 、dan bu he SAxiong D1ji yin qi dong zi zhong CGCG boxjiang lie 、te yi de jie ge ;qi dong zi huo xing LUCbao gao fen xi biao ming CAMTA3zai zhi wu ti nei fu diao kong CBP60ghe SARD1shi ji yin biao da ,er Pep1he Xoojian ruo CAMTA3de zhe chong fu diao kong zuo yong 。shui ming CAMTA3zhi jie fu diao kong CBP60g、ke neng jian jie fu diao kong SARD1,er Pep1he Xooyi zhi CAMTA3gai diao kong gong neng 。zhe xie jie guo biao ming ,CAMTA3tong guo zhi jie fu diao kong CBP60g、ke neng jian jie fu diao kong SARD1,yi zhi ICS1jie dao de SAge cheng ,cong er fu diao kong kang bing xing ;er Pep1he Xootong guo yi zhi CAMTA3gai gong neng cong er you dao kang bing xing 。(2)ming que le gai fei wai shi dui he pan jun kang xing de you dao zuo yong ji diao kong fen zi ji zhi 。gai fei (you xiao cheng fen wei CaCl2,0.3 mg/ml)xie mian pen wu xian zhe you dao ni na gai he you cai chan sheng dui he pan jun de kang xing ,kang xing chi xu ji zai 5tian yi shang 。tong yang gai fei chu li bu neng you dao cbp60g-1he sard1-1tu bian ti chan sheng zhe chong kang xing 。LUCbao gao fen xi xian shi ,gai fei yi zhi le CAMTA3dui CBP60ghe SARD1de fu diao kong zuo yong ,dan bu xian zhe gai bian CAMTA3ji yin biao da ,biao ming gai fei tong guo yi zhi CAMTA3dan bai dui CBP60ghe SARD1de fu diao kong zuo yong ,shi fang CBP60ghe SARD1,cong er ji fa kang bing xing 。(3)jian ding huo de le mo li suan (Jasmonic acid,JA)、huo xing yang (Reactive oxygen species,ROS)he RNAgan rao (RNA intefering,RNAi)tong lu ji yin zhong de CAMTA3ba biao ji yin ,jie shi CAMTA3can yu diao kong zhe xie tong lu 。qi dong zi xu lie jie guo xian shi ,JAtu jing ji yin JAZ1、JAZ2、JAZ5、JAZ7、JAZ8he MYC2、ROS tong lu NADPH yang hua mei ji yin RbohF yi ji RNAitong lu ji yin AGO4 AGO5、AGO6、RDR1、RDR3、RDR4de qi dong zi han you CGCGbox。EMSAfen xi biao ming ni na gai CAMTA3 CG-1jie gou yu zai ti wai zhi jie jie ge AGO4、AGO5、AGO6、RDR3、RBOHF、JAZ1he MYC2de qi dong zi ou CGCG box。qi dong zi huo xing LUCbao gao fen xi fa xian zai zhi wu ti nei ni na gai CAMTA3zheng diao kong AGO5he JAZ1de zhuai lu biao da 。zeng shang suo shu ,ben yan jiu chan ming le ni na gai CAMTA3dui he pan jun de ji chu kang xing he you dao kang xing yi ji dui Xoode fei ji zhu kang xing de an pu diao kong gong neng ,jie shi le ji yu CAMTA3yi CBP60ghe SARD1wei ba biao de SAge cheng diao kong xin tu jing ,ming que le yi gai tu jing wei ji chu de an pu kang bing diao kong xin ji zhi 。jian ding le JA、ROShe RNAitong lu zhong de CAMTA3ba biao ,jie shi le CAMTA3dui zhe xie tong lu de diao kong zuo yong 。ming que le gai fei dui he pan jun kang xing de you dao zuo yong ,jie shi le gai fei tong guo yi zhi CAMTA3dui CBP60ghe SARD1de fu diao kong zuo yong ji fa kang bing xing de fen zi ji zhi 。zhe xie zuo yong he ji zhi de jie shi zeng jin le zhi wu kang bing fen zi ji zhi de li jie 。Pep1he gai fei you kang zuo yong de fa xian ,hai wei jun he bing fang kong di gong qian zai de xin fang fa 。
论文参考文献
论文详细介绍
论文作者分别是来自浙江大学的徐雅静，发表于刊物浙江大学2019-05-20论文，是一篇关于钙肥论文,抗病性论文,核盘菌论文,稻白叶枯病菌论文,分子机制论文,水杨酸论文,茉莉酸论文,活性氧论文,浙江大学2019-05-20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，资料来自浙江大学2019-05-20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：钙肥论文; 抗病性论文; 核盘菌论文; 稻白叶枯病菌论文; 分子机制论文; 水杨酸论文; 茉莉酸论文; 活性氧论文; 浙江大学2019-05-20论文;