阿维菌素亚致死剂量对巴氏新小绥螨不同温度品系实验种群影响研究

发布时间：2018-06-19 20:28

本文选题：巴氏新小绥螨 + 阿维菌素　；参考：《西南大学》2017年硕士论文

【摘要】：巴氏新小绥螨Neoseiulus barkeri(Hughes),隶属于蛛形纲(Arachnida)、蜱螨亚纲(Acari)、寄螨目(Parasitiformes)、植绥螨科(Phytoseiidae),可以捕食叶螨、蓟马等小型害虫,是生物防治非常重要的天敌之一。本文以巴氏新小绥螨常温品系和耐高温品系为研究对象,在测定了阿维菌素对巴氏新小绥螨常温品系和耐高温品系毒力的基础上,开展了阿维菌素亚致死剂量胁迫对巴氏新小绥螨两种温度品系捕食功能的影响、两种温度品系体内主要解毒代谢酶的影响(包括剂量效应和时间效应)以及10个解毒代谢基因对阿维菌素胁迫响应的浓度效应和时间效应等方面的研究。主要研究内容和结果如下:1.阿维菌素对巴氏新小绥螨两种温度品系及二斑叶螨(Tetranychus urticae)的室内毒力测定采用离心管药膜法对巴氏新小绥螨两种温度品系以及靶标猎物二斑叶螨的雌成螨进行了室内毒力测定。结果表明阿维菌素对巴氏新小绥螨常温品系的LC10、LC20、LC30、LC50分别为1066.42mg/L、1739.56mg/L、2475.57mg/L、4435.98mg/L,阿维菌素对巴氏新小绥螨耐高温品系的LC10、LC20、LC30、LC50分别为565.10mg/L、910.08mg/L、1283.23mg/L、2264.59mg/L,阿维菌素对二斑叶螨的LC50为0.355 mg/L。2.阿维菌素亚致死剂量对巴氏新小绥螨两种温度品系捕食功能的影响本实验主要研究了阿维菌素三个亚致死剂量LC10、LC20、LC30处理后的巴氏新小绥螨常温品系和耐高温品系对二斑叶螨卵的捕食作用。结果表明阿维菌素不同亚致死剂量处理后,巴氏新小绥螨两种温度品系对二斑叶螨卵的捕食量均下降,且随阿维菌素亚致死剂量的增加,影响作用也越大;相同级别的阿维菌素亚致死剂量处理下,巴氏新小绥螨常温品系的捕食量大于高耐温品系。巴氏新小绥螨两种温度品系对二斑叶螨的捕食功能反应模型均为HollingⅡ模型,两个品系分别与各自的对照相比,各亚致死剂量处理下的瞬时攻击系数(6减小,处理猎物的时间?延长,捕食能力(6/?降低,最大日捕食量1/?降低,从而降低了巴氏新小绥螨对猎物的控制作用。3.阿维菌素亚致死剂量对巴氏新小绥螨两种温度品系主要解毒酶活性的影响本实验研究了阿维菌素亚致死剂量处理,对巴氏新小绥螨两种温度品系体内GSTs、MFOs和Car Es等解毒酶活性影响的剂量效应和时间效应。结果表明阿维菌素亚致死剂量对巴氏新小绥螨两种温度品系GSTs活性均有一定的抑制作用,存在一定的剂量效应和时间效应,即剂量越大,相同浓度处理时间越长,抑制作用也越强,且对耐高温品系的抑制作用较常温品系更强。阿维菌素亚致死剂量对巴氏新小绥螨两种温度品系MFOs和Car Es活性均有一定的诱导作用。具体表现为在阿维菌素浓度相对较低时,其对巴氏新小绥螨两种温度品系MFOs和Car Es的诱导作用较强,活性升高,以抵御外界不良环境,而后随着浓度增大,诱导作用减弱;阿维菌素LC30处理时间越长,对耐高温品系MFOs和Car Es的诱导作用越强,但对常温品系MFOs和Car Es的诱导作用则会有所减弱。4.巴氏新小绥螨两种温度品系阿维菌素胁迫下解毒代谢基因的表达模式分析本实验从巴氏新小绥螨转录组数据中找出获得注释的三种解毒代谢基因,对其进行系统进化分析后,利用内参基因(Beta-actin)作为参照,使用q PCR技术对解毒代谢基因进行验证,研究分析巴氏新小绥螨常温品系和耐高温品系10个解毒代谢基因(4个GSTs基因、4个MFOs基因以及2个Car Es基因)对阿维菌素胁迫响应的浓度效应和时间效应,明确了这10个解毒代谢基因在阿维菌素诱导后的m RNA表达量的变化趋势,以期为培育抗药品系的巴氏新小绥螨提供参考。结果表明巴氏新小绥螨常温品系:GST1、GST2、CYP3和CYP4的相对表达量在不同浓度以及LC30处理不同时间后均下调,但下调幅度有所差别;GST3在阿维菌素剂量较小时其相对表达量有所下调,剂量较大时其相对表达量上调,其在LC30处理不同时间后相对表达量均上调,但上调的幅度随着时间的延长会有所变化;GST4在阿维菌素各剂量处理后,其相对表达量均上调,但上调的幅度有所差别,其在阿维菌素LC30处理6h和24h后相对表达量均上调,在LC30处理12h后相对表达量下调;CYP1的相对表达量随着阿维菌素剂量的增大而增大,其在LC30处理6h和12h后相对表达量均下调,但在处理24h后,相对表达量却表现上调;CYP2的相对表达量在LC10、LC30处理组有所下调,在LC50处理组却表现上调,其在LC30处理不同时间后相对表达量全部下调;Car E1和Car E2的相对表达量均在阿维菌素剂量较小时表现上调,随着剂量加大,其上调幅度减小或甚至下调,LC30胁迫初期(6h)有所上调,随着胁迫时间延长,其相对表达量下调,但随后又有所回升。表明GST1、GST2、CYP3和CYP4可能不参与巴氏新小绥螨常温品系对阿维菌素的解毒代谢;而GST3、GST4、CYP1、CYP2、Car E1和Car E2可能都有参与巴氏新小绥螨常温品系对阿维菌素的解毒代谢,因而它们也可能参与巴氏新小绥螨常温品系对阿维菌素的抗性形成。巴氏新小绥螨耐高温品系:GST1、GST2的相对表达量在阿维菌素剂量较小时表现上调,但幅度有所差别,剂量达到LC50时却表现下调,其在阿维菌素LC30处理不同时间后相对表达量均上调,但幅度有所差别,于12h处理组达到最大;GST3的相对表达量在阿维菌素剂量较小时表现上调,但幅度有所差别,剂量达到LC50时却下调了,其在阿维菌素LC30处理6h和12h后有所下调,处理24h后却上调了;GST4在阿维菌素各剂量处理后,其相对表达量均上调,且随着阿维菌素剂量的增大,其上调幅度也变大,其在阿维菌素LC30处理不同时间后相对表达量均上调,但幅度有所差别,于12h处理组达到最大;CYP1在阿维菌素各剂量处理后,其相对表达量均上调,随着剂量增大上调幅度有所下降,其在阿维菌素LC30处理不同时间后相对表达量均上调,但均无显著性差异;CYP2在阿维菌素LC10处理后,其相对表达量上调,随着剂量增大,相对表达量却下调了,其在阿维菌素LC30处理6h后上调至对照的7.27倍,随着处理时间的延长,其相对表达量的上调幅度减小甚至下调;CYP3、CYP4、Car E1和Car E2的相对表达量均在阿维菌素剂量较小时表现上调,随着剂量加大,其上调幅度减小或甚至下调,其在阿维菌素LC30处理不同时间后均上调,但上调幅度先增大后减小,于6h处理组达到最大。表明GST1、GST2、GST3、GST4、CYP1、CYP2、CYP3、CYP4、Car E1和Car E2可能都有参与巴氏新小绥螨耐高温品系对阿维菌素的解毒代谢,因而它们也可能参与巴氏新小绥螨耐高温品系对阿维菌素的抗性形成。通过以上几个方面的研究,不仅可为巴氏新小绥螨常温品系和耐高温品系的田间释放提供一定的参考,还能判断解毒酶系以及某一解毒代谢基因在巴氏新小绥螨两种温度品系体内所起的作用,为巴氏新小绥螨抗阿维菌素品系的筛选提供参考依据。
[Abstract]:Neoseiulus barkeri (Hughes) of the new small suivert mite, belonging to the arachnoid (Arachnida), the acaroid Acara (Acari), the maidacara (Parasitiformes), and the family of the family of the family (Phytoseiidae). It is one of the most important natural enemies in biological control, which is one of the most important natural enemies of biological control. This paper is based on the study of the normal temperature strain and high temperature resistant strain of the new small suivert mite. Object. On the basis of measuring the virulence of abamectin to the normal temperature strain and high temperature resistant strain of mite pasteuri, the effects of Avi Randsuya lethal dose stress on the predatory function of two temperature strains of mite pasteurus pasteuri were carried out. The effects of the main detoxification and metabolic enzymes (including dose effect and time effect) on the main detoxification enzymes in the two kinds of temperature strain were also carried out. Studies on the concentration effect and time effect of 10 detoxification metabolic genes on the response to abamectin stress. The main contents and results are as follows: 1. the indoor virulence of two species of Acara pasteuriaris and the Tetranychus urticae in the Acara pasteuri, two kinds of temperature of the new small suipid mites of pasteurus pasteuri by centrifuge tube medicine membrane method The results showed that the LC10, LC20, LC30, and LC50 of abamectin were 1066.42mg/L, 1739.56mg/L, 2475.57mg/L, 4435.98mg/L, LC10, LC20, LC30, and 565.1, respectively, for the high temperature resistant strains of the new mite of pasteurus pasteuri, respectively, 565.1, respectively. The effects of 0mg/L, 910.08mg/L, 1283.23mg/L, 2264.59mg/L, Abamectin on the predation function of the sublethal dose of 0.355 mg/L.2. Abamectin on the LC50 of the two leaf mite to the two temperature strains of the new suikus pasteurioris, the experiment mainly studied the three sublethal doses of abamectin, LC10, LC20, and LC30, and the resistance to the mite. The results showed that after treatment with different lethal doses of avermectin, the predation of two temperature strains of Acara pasteuri was decreased with the increase of the lethal dose of Avi Randsuya, and the effect was greater with the increase of Avi Randsuya lethal dose; under the same level of sublethal dose of abamectin, BA. The predation rate of the normal temperature strain of suistee mite was greater than that of the high temperature resistant strain. The predatory response model of the two strains of mite pasteurus pasteuri was Holling II model, and the two strains were compared with their respective controls. The number of instantaneous attack lines under each sublethal dose (6 decreased, the time of handling the prey was prolonged, and the predation was prolonged. Capacity (6/? Reduction, maximum daily feed 1/? Reduction, thus reducing the control effect of a new pasteurus mite to prey.3. sublethal dose of abamectin to the main detoxification enzyme activities of the two species of pasteurus suikus The dose effect and time effect of the detoxification enzyme activities such as STs, MFOs and Car Es. The results showed that the sublethal dose of abamectin had a certain inhibitory effect on the activity of GSTs in the two strains of mite pasteuri. There was a certain dose effect and time effect, that is, the greater the dose, the longer the treatment time of the same concentration, the stronger the inhibition effect. The sublethal doses of abamectin have a certain induction effect on the activity of two kinds of temperature strain MFOs and Car Es of the mite of pasteurus pasteuri. The specific expression is that, when the abamectin concentration is relatively low, the induction effect on the two temperature strains of the mite pasteuri, MFOs and Car Es, is relatively low. The stronger and more active to resist the outside environment, and then decrease the induction effect with the increase of concentration, the longer the treatment time of abamectin LC30, the stronger the induction of MFOs and Car Es in high temperature resistant strain, but the induction of MFOs and Car Es on the normal temperature strain will weaken the Abamectin stress of the two temperature strains of the new small suipah mites of.4. Bart. Analysis of the expression pattern of detoxification metabolic genes, three detoxifying metabolic genes were found from the data of the new small suipid mite transcriptional group. After the phylogenetic analysis, the Beta-actin was used as a reference, and the Q PCR technique was used to verify the detoxification gene, and the new small suivert mite was studied and analyzed. The concentration effect and time effect of 10 detoxifying metabolic genes (4 GSTs genes, 4 MFOs genes and 2 Car Es genes) on the response of avermectin stress, and the change trend of the m RNA expression of the 10 detoxifying metabolic genes after abamectin induction was defined. The results showed that the relative expressions of GST1, GST2, CYP3 and CYP4 were down regulated at different concentrations and after LC30 treatment at different times, but the decrease was different, and the relative expression of GST3 at the lower dose of abamectin was lower, and the relative expression was up to be up when the dose was larger, and it was in LC30. The relative expression was up up after treatment at different time, but the range of up-regulation was changed with time. The relative expression of GST4 was up up after all doses of avermectin treatment, but the range of up regulation was different. The relative expression was up up after abamectin LC30 treatment of 6h and 24h, and the relative expression after LC30 treatment of 12h was under the relative expression. The relative expression of CYP1 increased with the increase of the avermectin dose, and its relative expression decreased after the LC30 treatment of 6h and 12h, but the relative expression increased after 24h treatment. The relative expression of CYP2 was in LC10, LC30 treatment group was down, and the relative expression was up in the LC50 processing group, and the relative expression after LC30 was treated at different time. The relative expression of Car E1 and Car E2 up-regulated at the lower dose of abamectin, as the dosage increased, the up-regulation amplitude decreased or even down, and the LC30 stress early (6h) up up. As the time of stress prolonged, its relative expression decreased, but then rebounded. It indicated that GST1, GST2, CYP3 and CYP4 may not. GST3, GST4, CYP1, CYP2, Car E1 and Car E2 may be involved in the detoxification of abamectin, and they may also be involved in the resistance to avermectin. The relative expression of GST1, GST2 was up-regulated at the low dose of abamectin, but the amplitude was different. The relative expression was down when the dose reached LC50. The relative expression of the abamectin LC30 was up, but the amplitude was different in the 12h treatment group. The relative expression of GST3 in the abamectin dose was more than that of the abamectin. The hourly performance was up, but the amplitude was different. When the dose reached LC50, it was down. After the abamectin LC30 was treated with 6h and 12h, it was down regulated. After treatment of 24h, the relative expression of GST4 was up, and its up range increased with the increase of abamectin, and it was in the abamectin LC3. 0 at different time, the relative expression was up up, but the amplitude was different, and the 12h treatment group reached the maximum. After all doses of abamectin were treated, the relative expression of CYP1 was up, and the relative expression of the abamectin LC30 was up up, but there was no significant difference. After the treatment of abamectin LC10, the relative expression of CYP2 was up. As the dose increased, the relative expression was down. The relative expression was up to 7.27 times after the abamectin LC30 was treated with 6h. The relative expression of the relative expression decreased and even decreased with the prolongation of the treatment time. The relative expressions of CYP3, CYP4, Car E1 and Car E2 were all in avi. The amount of bacteriocin was up-regulated in the lower dose. As the dose increased, its up range decreased or even downregulated. It was up up after the treatment of abamectin LC30 at different times, but the up range increased first and then decreased, and reached the maximum in the 6h treatment group. It indicated that GST1, GST2, GST3, GST4, CYP1, CYP2, CYP3, CYP4, Car E1, and CYP4 were probably involved in pasteurized new small pacing. They may also participate in the resistance formation of abamectin to the high temperature resistant strain of the mite. Through the study above, it can not only provide some reference for the field release of the normal temperature strain and high temperature resistant strain of pasteurus pasteuriaris, but also can judge the detoxification enzyme system. The function of a detoxifying metabolic gene in two temperature strains of the new suibitus pasteuriaris, which provides a reference for the screening of the anti abamectin strains of the new small suipaiacara pasteuri.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S476.2

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