本文主要研究内容
作者侯佳音（2019）在《番茄TGA2转录因子在农药百菌清代谢降解中的功能及调控机制》一文中研究指出：设施农业的兴起和快速发展,解决了我国长期以来蔬菜供应不足的困境,但也不可避免地带来了一些问题,如设施内特定环境引起的病虫害高发和连作障碍,以及由于农药过量施用引起的农药残留超标等。因此,寻求一种绿色高效、简便易行且能够大规模应用的农药残留消除技术和方案显得尤为迫切。油菜素内酯(Brassinosteroids,BRs)是一种调控植物生长发育和抗逆性的新型植物激素,并且能够通过激活解毒代谢基因促进异源化合物降解,但内在的调控机制尚不明确。TGA转录因子是bZIP转录因子(basic region-leucine zippers)家族成员,是植物中最早鉴定到的一类转录因子,因其结合基因启动子的TGACG序列而得名。TGA转录因子参与植物的抗病防御、生长发育以及对有毒代谢产物的解毒代谢等多个过程,但其在农药残留代谢降解中的作用尚未见报道。番茄是设施园艺栽培面积最大的蔬菜,本文选择番茄作为实验材料,利用转基因技术,结合分子生物学和植物生理学手段,研究TGA转录因子在BR促进广谱杀菌剂百菌清(Chlorothalonil,CHT)植物体内代谢降解的作用,并研究TGA2转录因子的氧化还原修饰对其调控解毒代谢基因表达和农药残留代谢降解的影响。结果如下:1.明确了番茄中有四个Class Ⅱ TGA转录因子,其中TGA2、TGA2.2和TGA2.3参与了番茄植株对百菌清残留的降解过程。TGA2蛋白能够直接结合谷胱甘肽S-转移酶(GlutathioneS-transferase,GST)编码基因GST3的启动子并促进其转录。利用拟南芥(Arabidopsis thaliana)TGA蛋白序列进行Blast比对,鉴定出番茄的10个TGA转录因子,其中Class Ⅱ TGAs中TGA2、TG2.2、TGA2.3基因沉默抑制CHT处理诱导的解毒基因表达和酶活性的提高以及谷胱甘肽含量的增加,进而导致CHT残留升高。TGA2基因过表达增强了植株对CHT的代谢降解能力。TGA2基因沉默导致解毒基因GST3的转录受到抑制,而过表达TGA2基因使GST3的基因表达显著提高。凝胶电泳迁移实验(EMSA)和染色质免疫共沉淀实验(ChIP-qPCR)结果表明,TGA2蛋白能够与GST3基因启动子结合,促进基因的转录表达。GST3基因沉默抑制了植株的解毒代谢,导致CHT残留量显著增加。2.BR诱导的质外体H2O2激活谷氧还蛋白GRXS13和GRXS25编码基因,促进两者与TGA2转录因子发生蛋白互作并共同参与CHT残留代谢降解。外源BR处理能够通过激活解毒代谢基因表达降低番茄植株CHT残留。TGA2基因沉默抑制了外源BR对植株CHT残留代谢降解的促进作用。通过比较呼吸爆发氧化酶同系物编码基因RBOH1沉默植株与对照植株,发现CC-type谷氧还蛋白基因GRXS13和GRXS25受BR和CHT诱导,但是其表达依赖于RBOH1介导的质外体H2O2。GRXS13和GRXS25基因沉默抑制BR诱导的CHT残留代谢降解。酵母双杂交及双分子荧光互补实验证明,GRXS13和GRXS25与TGA2存在蛋白水平的相互作用,揭示了 TGA2转录因子在BR诱导的质外体H2O2信号下游受到GRX介导的谷胱甘肽化修饰的可能。3.BR和CHT能够诱导植株S-亚硝基硫醇(SNO)累积,S-亚硝基谷胱甘肽(GSNO)作为一氧化氮(NO)供体促进TGA2蛋白S-亚硝基化修饰,进而增强其转录活性、解毒代谢基因表达和农残代谢降解。BR和CHT通过诱导硝酸还原酶编码基因NR转录水平表达,增加叶片SNO含量。亚硝基谷胱甘肽还原酶(GSNOR)能够通过抑制GSNO的积累,降低SNO的水平。沉默GSNOR基因提高了 GSTs的转录水平和酶活性,降低了植株中CHT的残留。体外实验表明,GSNO能够诱导TGA2蛋白发生S-亚硝基化修饰,这种修饰增强了 TGA2蛋白对解毒基因GST3启动子的结合能力。以上结果说明,番茄对CHT解毒代谢过程中,TGA2蛋白的S-亚硝基化修饰对其转录活性及解毒代谢功能有重要的调控作用。
Abstract
she shi nong ye de xing qi he kuai su fa zhan ,jie jue le wo guo chang ji yi lai shu cai gong ying bu zu de kun jing ,dan ye bu ke bi mian de dai lai le yi xie wen ti ,ru she shi nei te ding huan jing yin qi de bing chong hai gao fa he lian zuo zhang ai ,yi ji you yu nong yao guo liang shi yong yin qi de nong yao can liu chao biao deng 。yin ci ,xun qiu yi chong lu se gao xiao 、jian bian yi hang ju neng gou da gui mo ying yong de nong yao can liu xiao chu ji shu he fang an xian de you wei pai qie 。you cai su nei zhi (Brassinosteroids,BRs)shi yi chong diao kong zhi wu sheng chang fa yo he kang ni xing de xin xing zhi wu ji su ,bing ju neng gou tong guo ji huo jie du dai xie ji yin cu jin yi yuan hua ge wu jiang jie ,dan nei zai de diao kong ji zhi shang bu ming que 。TGAzhuai lu yin zi shi bZIPzhuai lu yin zi (basic region-leucine zippers)jia zu cheng yuan ,shi zhi wu zhong zui zao jian ding dao de yi lei zhuai lu yin zi ,yin ji jie ge ji yin qi dong zi de TGACGxu lie er de ming 。TGAzhuai lu yin zi can yu zhi wu de kang bing fang yu 、sheng chang fa yo yi ji dui you du dai xie chan wu de jie du dai xie deng duo ge guo cheng ,dan ji zai nong yao can liu dai xie jiang jie zhong de zuo yong shang wei jian bao dao 。fan jia shi she shi yuan yi zai pei mian ji zui da de shu cai ,ben wen shua ze fan jia zuo wei shi yan cai liao ,li yong zhuai ji yin ji shu ,jie ge fen zi sheng wu xue he zhi wu sheng li xue shou duan ,yan jiu TGAzhuai lu yin zi zai BRcu jin an pu sha jun ji bai jun qing (Chlorothalonil,CHT)zhi wu ti nei dai xie jiang jie de zuo yong ,bing yan jiu TGA2zhuai lu yin zi de yang hua hai yuan xiu shi dui ji diao kong jie du dai xie ji yin biao da he nong yao can liu dai xie jiang jie de ying xiang 。jie guo ru xia :1.ming que le fan jia zhong you si ge Class Ⅱ TGAzhuai lu yin zi ,ji zhong TGA2、TGA2.2he TGA2.3can yu le fan jia zhi zhu dui bai jun qing can liu de jiang jie guo cheng 。TGA2dan bai neng gou zhi jie jie ge gu guang gan tai S-zhuai yi mei (GlutathioneS-transferase,GST)bian ma ji yin GST3de qi dong zi bing cu jin ji zhuai lu 。li yong ni na gai (Arabidopsis thaliana)TGAdan bai xu lie jin hang Blastbi dui ,jian ding chu fan jia de 10ge TGAzhuai lu yin zi ,ji zhong Class Ⅱ TGAszhong TGA2、TG2.2、TGA2.3ji yin chen mo yi zhi CHTchu li you dao de jie du ji yin biao da he mei huo xing de di gao yi ji gu guang gan tai han liang de zeng jia ,jin er dao zhi CHTcan liu sheng gao 。TGA2ji yin guo biao da zeng jiang le zhi zhu dui CHTde dai xie jiang jie neng li 。TGA2ji yin chen mo dao zhi jie du ji yin GST3de zhuai lu shou dao yi zhi ,er guo biao da TGA2ji yin shi GST3de ji yin biao da xian zhe di gao 。ning jiao dian yong qian yi shi yan (EMSA)he ran se zhi mian yi gong chen dian shi yan (ChIP-qPCR)jie guo biao ming ,TGA2dan bai neng gou yu GST3ji yin qi dong zi jie ge ,cu jin ji yin de zhuai lu biao da 。GST3ji yin chen mo yi zhi le zhi zhu de jie du dai xie ,dao zhi CHTcan liu liang xian zhe zeng jia 。2.BRyou dao de zhi wai ti H2O2ji huo gu yang hai dan bai GRXS13he GRXS25bian ma ji yin ,cu jin liang zhe yu TGA2zhuai lu yin zi fa sheng dan bai hu zuo bing gong tong can yu CHTcan liu dai xie jiang jie 。wai yuan BRchu li neng gou tong guo ji huo jie du dai xie ji yin biao da jiang di fan jia zhi zhu CHTcan liu 。TGA2ji yin chen mo yi zhi le wai yuan BRdui zhi zhu CHTcan liu dai xie jiang jie de cu jin zuo yong 。tong guo bi jiao hu xi bao fa yang hua mei tong ji wu bian ma ji yin RBOH1chen mo zhi zhu yu dui zhao zhi zhu ,fa xian CC-typegu yang hai dan bai ji yin GRXS13he GRXS25shou BRhe CHTyou dao ,dan shi ji biao da yi lai yu RBOH1jie dao de zhi wai ti H2O2。GRXS13he GRXS25ji yin chen mo yi zhi BRyou dao de CHTcan liu dai xie jiang jie 。jiao mu shuang za jiao ji shuang fen zi ying guang hu bu shi yan zheng ming ,GRXS13he GRXS25yu TGA2cun zai dan bai shui ping de xiang hu zuo yong ,jie shi le TGA2zhuai lu yin zi zai BRyou dao de zhi wai ti H2O2xin hao xia you shou dao GRXjie dao de gu guang gan tai hua xiu shi de ke neng 。3.BRhe CHTneng gou you dao zhi zhu S-ya xiao ji liu chun (SNO)lei ji ,S-ya xiao ji gu guang gan tai (GSNO)zuo wei yi yang hua dan (NO)gong ti cu jin TGA2dan bai S-ya xiao ji hua xiu shi ,jin er zeng jiang ji zhuai lu huo xing 、jie du dai xie ji yin biao da he nong can dai xie jiang jie 。BRhe CHTtong guo you dao xiao suan hai yuan mei bian ma ji yin NRzhuai lu shui ping biao da ,zeng jia xie pian SNOhan liang 。ya xiao ji gu guang gan tai hai yuan mei (GSNOR)neng gou tong guo yi zhi GSNOde ji lei ,jiang di SNOde shui ping 。chen mo GSNORji yin di gao le GSTsde zhuai lu shui ping he mei huo xing ,jiang di le zhi zhu zhong CHTde can liu 。ti wai shi yan biao ming ,GSNOneng gou you dao TGA2dan bai fa sheng S-ya xiao ji hua xiu shi ,zhe chong xiu shi zeng jiang le TGA2dan bai dui jie du ji yin GST3qi dong zi de jie ge neng li 。yi shang jie guo shui ming ,fan jia dui CHTjie du dai xie guo cheng zhong ,TGA2dan bai de S-ya xiao ji hua xiu shi dui ji zhuai lu huo xing ji jie du dai xie gong neng you chong yao de diao kong zuo yong 。
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论文作者分别是来自浙江大学的侯佳音，发表于刊物浙江大学2019-09-18论文，是一篇关于百菌清残留论文,翻译后修饰论文,油菜素内酯论文,谷氧还蛋白论文,亚硝基化论文,番茄论文,浙江大学2019-09-18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2019-09-18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：百菌清残留论文; 翻译后修饰论文; 油菜素内酯论文; 谷氧还蛋白论文; 亚硝基化论文; 番茄论文; 浙江大学2019-09-18论文;