本文主要研究内容
作者李金航（2019）在《丁氟螨酯对朱砂叶螨和巴氏新小绥螨的选择性机制》一文中研究指出：朱砂叶螨（Tetranychus cinnabarinus）是一种世界范围内的重要农业害螨,可危害豇豆、茄子、棉花、花卉等多种农业、经济作物。由于杀螨剂大量持续不科学使用以及害螨具繁殖能力强、世代周期短等特点,朱砂叶螨的抗药性问题日趋严重。巴氏新小绥螨（Neoseiulus barkeri）具有发育历期短、产卵率高等优点,主要用于防治为害果树、蔬菜和棉花等作物的害螨、蓟马等小型害虫,目前巴氏新小绥螨已经实现规模化饲养,是生物防治最佳天敌之一。丁氟螨酯是一种苯酰乙腈类触杀型杀螨剂,其作用靶标为线粒体复合物Ⅱ,具有生物活性高、持效性好、对非靶标生物安全等优点。本论文以朱砂叶螨和巴氏新小绥螨为研究对象,在室内确定丁氟螨酯对朱砂叶螨和巴氏新小绥螨毒力存在上万倍差异的基础上,通过室内生物测定、酶活测定、基因克隆和序列比对等方法着重研究了丁氟螨酯对朱砂叶螨和巴氏新小绥螨毒力差异的生理生化及分子机制,主要研究内容和结果如下:1、杀螨剂对朱砂叶螨和巴氏新小绥螨毒力差异研究采用药膜法对朱砂叶螨和巴氏新小绥螨进行了室内生物测定。结果表明丁氟螨酯对朱砂叶螨的LC50为7.816 mg/L,对巴氏新小绥螨的LC50超过了100,000mg/L（无法测出具体数值）,捕食螨的不敏感性超过叶螨的12,794倍;腈吡螨酯对朱砂叶螨的LC50为4.587 mg/L,对巴氏新小绥螨的LC50超过了50,000 mg/L（无法测出具体数值）,捕食螨的不敏感性超过叶螨的10,900倍;对照药剂甲氰菊酯对朱砂叶螨和巴氏新小绥螨的LC50分别为606.98 mg/L和140.923 mg/L,捕食螨的不敏感性为叶螨的0.23倍;马拉硫磷对朱砂叶螨和巴氏新小绥螨的LC50分别为4112.088 mg/L和65.594 mg/L,后者的不敏感性为前者的0.02倍。三种增效剂PBO、DEM和TPP分别单独与丁氟螨酯混用对朱砂叶螨的增效比分别为1.43、1.86和2.33,而三种增效剂PBO、DEM和TPP分别单独与丁氟螨酯混用,对巴氏新小绥螨的LC50仍然大于100,000 mg/L,无法计算出增效比。生物测定结果表明丁氟螨酯对朱砂叶螨和巴氏新小绥螨存在巨大的毒力差异,且对照药剂与增效剂生物测定结果间接表明表皮穿透能力以及代谢酶活性并不是导致丁氟螨酯对叶螨和捕食螨毒力差异的主要原因,并且强烈暗示靶标差异是SQR抑制剂类杀螨剂在叶螨和捕食螨间具有优异选择性的主要原因。2、朱砂叶螨和巴氏新小绥螨解毒酶活性研究在增效剂实验的基础上,进一步检测、比较了朱砂叶螨和巴氏新小绥螨三大解毒酶的活性差异。结果显示朱砂叶螨谷胱甘肽-S-转移酶（GSTs）和细胞色素P450酶（P450s）的活性均显著高于巴氏新小绥螨,羧酸酯酶（CarEs）活性显著低于巴氏新小绥螨。确定丁氟螨酯对朱砂叶螨的LC10、LC30和LC50为诱导处理浓度,朱砂叶螨经不同浓度(LC10、LC30和LC50)丁氟螨酯诱导后酶活性变化如下:羧酸酯酶活性经LC30诱导后显著增加,谷胱甘肽-S-转移酶活性经LC50和LC30诱导后显著降低,其余诱导处理酶活性没有显著变化。巴氏新小绥螨经丁氟螨酯LC50和LC30诱导后P450酶活性显著上升,其余诱导处理酶活性无显著变化。解毒酶活性检测比较结果说明,巴氏新小绥螨的三大解毒酶活性及其响应丁氟螨酯胁迫能力并不比朱砂叶螨更强,反过来说明巴氏新小绥螨对丁氟螨酯的耐药力超过朱砂叶螨12,794倍的原因与解毒代谢酶活性差异关系不大。3、朱砂叶螨和巴氏新小绥螨线粒体复合物Ⅱ活性及ATP含量测定对朱砂叶螨和巴氏新小绥螨的线粒体电子传递链复合物Ⅱ（SQR,丁氟螨酯的作用靶标）活性测定表明:巴氏新小绥螨的SQR活性为朱砂叶螨SQR活性的1.63倍。丁氟螨酯对朱砂叶螨和巴氏新小绥螨离体SQR活性抑制中浓度(IC50)分别为23.141 nmol/L和4,531,160 nmol/L,巴氏新小绥螨SQR对丁氟螨酯的不敏感性(N.b IC50/T.c IC50)是朱砂叶螨的195,807倍,强烈暗示出丁氟螨酯对朱砂叶螨和巴氏新小绥螨的毒力差异与其在二者体内作用靶标的差异有关。线粒体复合物Ⅱ（SQR）的主要生物功能是参与电子传递生成ATP,笔者使用ATP检测试剂盒检测、比较了朱砂叶螨和巴氏新小绥螨的ATP含量,结果显示朱砂叶螨的ATP含量为4.376 nmol/mg Pro,巴氏新小绥螨的ATP含量为13.594nmol/mg Pro。用10 mg/L的丁氟螨酯分别处理朱砂叶螨和巴氏新小绥螨4 h后,朱砂叶螨体内ATP含量显著下降,而巴氏新小绥螨ATP含量并无显著变化,这进一步说明由于朱砂叶螨和巴氏新小绥螨SQR结构的差异,导致丁氟螨酯在两种螨体内与作用靶标的结合抑制能力不同,从而表现为活性差异。4、朱砂叶螨和巴氏新小绥螨线粒体复合物Ⅱ基因克隆及活性位点分析通过对转录组数据进行分析,获得了朱砂叶螨和巴氏新小绥螨SQR基因片段。根据所获得基因片段,利用RACE（rapid-amplification of cDNA ends）技术,分别克隆获得了巴氏新小绥螨和朱砂叶螨SQR的5个亚基基因,分别命名为NbSDHA、NbSDHB、NbSDHC、NbSDHD、NbSDH5和TcSDHA、TcSDHB、TcSDHC、TcSDHD、TcSDH5。通过NCBI BLAST得到各亚基功能区域,对各亚基功能区域进行氨基酸序列比对,发现存在多达220个差异位点。这些差异位点一方面说明朱砂叶螨和巴氏新小绥螨的SQR结构存在较大差异,另一方面说明这些差异可能是丁氟螨酯对朱砂叶螨和巴氏新小绥螨存在优异选择性的主要原因。
Abstract
zhu sha xie man （Tetranychus cinnabarinus）shi yi chong shi jie fan wei nei de chong yao nong ye hai man ,ke wei hai jiang dou 、jia zi 、mian hua 、hua hui deng duo chong nong ye 、jing ji zuo wu 。you yu sha man ji da liang chi xu bu ke xue shi yong yi ji hai man ju fan shi neng li jiang 、shi dai zhou ji duan deng te dian ,zhu sha xie man de kang yao xing wen ti ri qu yan chong 。ba shi xin xiao sui man （Neoseiulus barkeri）ju you fa yo li ji duan 、chan luan lv gao deng you dian ,zhu yao yong yu fang zhi wei hai guo shu 、shu cai he mian hua deng zuo wu de hai man 、ji ma deng xiao xing hai chong ,mu qian ba shi xin xiao sui man yi jing shi xian gui mo hua si yang ,shi sheng wu fang zhi zui jia tian di zhi yi 。ding fu man zhi shi yi chong ben xian yi jing lei chu sha xing sha man ji ,ji zuo yong ba biao wei xian li ti fu ge wu Ⅱ,ju you sheng wu huo xing gao 、chi xiao xing hao 、dui fei ba biao sheng wu an quan deng you dian 。ben lun wen yi zhu sha xie man he ba shi xin xiao sui man wei yan jiu dui xiang ,zai shi nei que ding ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man du li cun zai shang mo bei cha yi de ji chu shang ,tong guo shi nei sheng wu ce ding 、mei huo ce ding 、ji yin ke long he xu lie bi dui deng fang fa zhao chong yan jiu le ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man du li cha yi de sheng li sheng hua ji fen zi ji zhi ,zhu yao yan jiu nei rong he jie guo ru xia :1、sha man ji dui zhu sha xie man he ba shi xin xiao sui man du li cha yi yan jiu cai yong yao mo fa dui zhu sha xie man he ba shi xin xiao sui man jin hang le shi nei sheng wu ce ding 。jie guo biao ming ding fu man zhi dui zhu sha xie man de LC50wei 7.816 mg/L,dui ba shi xin xiao sui man de LC50chao guo le 100,000mg/L（mo fa ce chu ju ti shu zhi ）,bu shi man de bu min gan xing chao guo xie man de 12,794bei ;jing bi man zhi dui zhu sha xie man de LC50wei 4.587 mg/L,dui ba shi xin xiao sui man de LC50chao guo le 50,000 mg/L（mo fa ce chu ju ti shu zhi ）,bu shi man de bu min gan xing chao guo xie man de 10,900bei ;dui zhao yao ji jia qing ju zhi dui zhu sha xie man he ba shi xin xiao sui man de LC50fen bie wei 606.98 mg/Lhe 140.923 mg/L,bu shi man de bu min gan xing wei xie man de 0.23bei ;ma la liu lin dui zhu sha xie man he ba shi xin xiao sui man de LC50fen bie wei 4112.088 mg/Lhe 65.594 mg/L,hou zhe de bu min gan xing wei qian zhe de 0.02bei 。san chong zeng xiao ji PBO、DEMhe TPPfen bie chan du yu ding fu man zhi hun yong dui zhu sha xie man de zeng xiao bi fen bie wei 1.43、1.86he 2.33,er san chong zeng xiao ji PBO、DEMhe TPPfen bie chan du yu ding fu man zhi hun yong ,dui ba shi xin xiao sui man de LC50reng ran da yu 100,000 mg/L,mo fa ji suan chu zeng xiao bi 。sheng wu ce ding jie guo biao ming ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man cun zai ju da de du li cha yi ,ju dui zhao yao ji yu zeng xiao ji sheng wu ce ding jie guo jian jie biao ming biao pi chuan tou neng li yi ji dai xie mei huo xing bing bu shi dao zhi ding fu man zhi dui xie man he bu shi man du li cha yi de zhu yao yuan yin ,bing ju jiang lie an shi ba biao cha yi shi SQRyi zhi ji lei sha man ji zai xie man he bu shi man jian ju you you yi shua ze xing de zhu yao yuan yin 。2、zhu sha xie man he ba shi xin xiao sui man jie du mei huo xing yan jiu zai zeng xiao ji shi yan de ji chu shang ,jin yi bu jian ce 、bi jiao le zhu sha xie man he ba shi xin xiao sui man san da jie du mei de huo xing cha yi 。jie guo xian shi zhu sha xie man gu guang gan tai -S-zhuai yi mei （GSTs）he xi bao se su P450mei （P450s）de huo xing jun xian zhe gao yu ba shi xin xiao sui man ,suo suan zhi mei （CarEs）huo xing xian zhe di yu ba shi xin xiao sui man 。que ding ding fu man zhi dui zhu sha xie man de LC10、LC30he LC50wei you dao chu li nong du ,zhu sha xie man jing bu tong nong du (LC10、LC30he LC50)ding fu man zhi you dao hou mei huo xing bian hua ru xia :suo suan zhi mei huo xing jing LC30you dao hou xian zhe zeng jia ,gu guang gan tai -S-zhuai yi mei huo xing jing LC50he LC30you dao hou xian zhe jiang di ,ji yu you dao chu li mei huo xing mei you xian zhe bian hua 。ba shi xin xiao sui man jing ding fu man zhi LC50he LC30you dao hou P450mei huo xing xian zhe shang sheng ,ji yu you dao chu li mei huo xing mo xian zhe bian hua 。jie du mei huo xing jian ce bi jiao jie guo shui ming ,ba shi xin xiao sui man de san da jie du mei huo xing ji ji xiang ying ding fu man zhi xie pai neng li bing bu bi zhu sha xie man geng jiang ,fan guo lai shui ming ba shi xin xiao sui man dui ding fu man zhi de nai yao li chao guo zhu sha xie man 12,794bei de yuan yin yu jie du dai xie mei huo xing cha yi guan ji bu da 。3、zhu sha xie man he ba shi xin xiao sui man xian li ti fu ge wu Ⅱhuo xing ji ATPhan liang ce ding dui zhu sha xie man he ba shi xin xiao sui man de xian li ti dian zi chuan di lian fu ge wu Ⅱ（SQR,ding fu man zhi de zuo yong ba biao ）huo xing ce ding biao ming :ba shi xin xiao sui man de SQRhuo xing wei zhu sha xie man SQRhuo xing de 1.63bei 。ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man li ti SQRhuo xing yi zhi zhong nong du (IC50)fen bie wei 23.141 nmol/Lhe 4,531,160 nmol/L,ba shi xin xiao sui man SQRdui ding fu man zhi de bu min gan xing (N.b IC50/T.c IC50)shi zhu sha xie man de 195,807bei ,jiang lie an shi chu ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man de du li cha yi yu ji zai er zhe ti nei zuo yong ba biao de cha yi you guan 。xian li ti fu ge wu Ⅱ（SQR）de zhu yao sheng wu gong neng shi can yu dian zi chuan di sheng cheng ATP,bi zhe shi yong ATPjian ce shi ji he jian ce 、bi jiao le zhu sha xie man he ba shi xin xiao sui man de ATPhan liang ,jie guo xian shi zhu sha xie man de ATPhan liang wei 4.376 nmol/mg Pro,ba shi xin xiao sui man de ATPhan liang wei 13.594nmol/mg Pro。yong 10 mg/Lde ding fu man zhi fen bie chu li zhu sha xie man he ba shi xin xiao sui man 4 hhou ,zhu sha xie man ti nei ATPhan liang xian zhe xia jiang ,er ba shi xin xiao sui man ATPhan liang bing mo xian zhe bian hua ,zhe jin yi bu shui ming you yu zhu sha xie man he ba shi xin xiao sui man SQRjie gou de cha yi ,dao zhi ding fu man zhi zai liang chong man ti nei yu zuo yong ba biao de jie ge yi zhi neng li bu tong ,cong er biao xian wei huo xing cha yi 。4、zhu sha xie man he ba shi xin xiao sui man xian li ti fu ge wu Ⅱji yin ke long ji huo xing wei dian fen xi tong guo dui zhuai lu zu shu ju jin hang fen xi ,huo de le zhu sha xie man he ba shi xin xiao sui man SQRji yin pian duan 。gen ju suo huo de ji yin pian duan ,li yong RACE（rapid-amplification of cDNA ends）ji shu ,fen bie ke long huo de le ba shi xin xiao sui man he zhu sha xie man SQRde 5ge ya ji ji yin ,fen bie ming ming wei NbSDHA、NbSDHB、NbSDHC、NbSDHD、NbSDH5he TcSDHA、TcSDHB、TcSDHC、TcSDHD、TcSDH5。tong guo NCBI BLASTde dao ge ya ji gong neng ou yu ,dui ge ya ji gong neng ou yu jin hang an ji suan xu lie bi dui ,fa xian cun zai duo da 220ge cha yi wei dian 。zhe xie cha yi wei dian yi fang mian shui ming zhu sha xie man he ba shi xin xiao sui man de SQRjie gou cun zai jiao da cha yi ,ling yi fang mian shui ming zhe xie cha yi ke neng shi ding fu man zhi dui zhu sha xie man he ba shi xin xiao sui man cun zai you yi shua ze xing de zhu yao yuan yin 。
论文参考文献
论文详细介绍
论文作者分别是来自西南大学的李金航，发表于刊物西南大学2019-09-24论文，是一篇关于朱砂叶螨论文,巴氏新小绥螨论文,丁氟螨酯论文,毒力差异论文,西南大学2019-09-24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，资料来自西南大学2019-09-24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：朱砂叶螨论文; 巴氏新小绥螨论文; 丁氟螨酯论文; 毒力差异论文; 西南大学2019-09-24论文;