本文選題：巴氏新小綏螨 + 阿維菌素　； 參考：《西南大學(xué)》2017年碩士論文

【摘要】：巴氏新小綏螨Neoseiulus barkeri(Hughes),隸屬于蛛形綱(Arachnida)、蜱螨亞綱(Acari)、寄螨目(Parasitiformes)、植綏螨科(Phytoseiidae),可以捕食葉螨、薊馬等小型害蟲,是生物防治非常重要的天敵之一。本文以巴氏新小綏螨常溫品系和耐高溫品系為研究對(duì)象,在測(cè)定了阿維菌素對(duì)巴氏新小綏螨常溫品系和耐高溫品系毒力的基礎(chǔ)上,開展了阿維菌素亞致死劑量脅迫對(duì)巴氏新小綏螨兩種溫度品系捕食功能的影響、兩種溫度品系體內(nèi)主要解毒代謝酶的影響(包括劑量效應(yīng)和時(shí)間效應(yīng))以及10個(gè)解毒代謝基因?qū)Π⒕S菌素脅迫響應(yīng)的濃度效應(yīng)和時(shí)間效應(yīng)等方面的研究。主要研究內(nèi)容和結(jié)果如下:1.阿維菌素對(duì)巴氏新小綏螨兩種溫度品系及二斑葉螨(Tetranychus urticae)的室內(nèi)毒力測(cè)定采用離心管藥膜法對(duì)巴氏新小綏螨兩種溫度品系以及靶標(biāo)獵物二斑葉螨的雌成螨進(jìn)行了室內(nèi)毒力測(cè)定。結(jié)果表明阿維菌素對(duì)巴氏新小綏螨常溫品系的LC10、LC20、LC30、LC50分別為1066.42mg/L、1739.56mg/L、2475.57mg/L、4435.98mg/L,阿維菌素對(duì)巴氏新小綏螨耐高溫品系的LC10、LC20、LC30、LC50分別為565.10mg/L、910.08mg/L、1283.23mg/L、2264.59mg/L,阿維菌素對(duì)二斑葉螨的LC50為0.355 mg/L。2.阿維菌素亞致死劑量對(duì)巴氏新小綏螨兩種溫度品系捕食功能的影響本實(shí)驗(yàn)主要研究了阿維菌素三個(gè)亞致死劑量LC10、LC20、LC30處理后的巴氏新小綏螨常溫品系和耐高溫品系對(duì)二斑葉螨卵的捕食作用。結(jié)果表明阿維菌素不同亞致死劑量處理后,巴氏新小綏螨兩種溫度品系對(duì)二斑葉螨卵的捕食量均下降,且隨阿維菌素亞致死劑量的增加,影響作用也越大;相同級(jí)別的阿維菌素亞致死劑量處理下,巴氏新小綏螨常溫品系的捕食量大于高耐溫品系。巴氏新小綏螨兩種溫度品系對(duì)二斑葉螨的捕食功能反應(yīng)模型均為HollingⅡ模型,兩個(gè)品系分別與各自的對(duì)照相比,各亞致死劑量處理下的瞬時(shí)攻擊系數(shù)(6減小,處理獵物的時(shí)間?延長,捕食能力(6/?降低,最大日捕食量1/?降低,從而降低了巴氏新小綏螨對(duì)獵物的控制作用。3.阿維菌素亞致死劑量對(duì)巴氏新小綏螨兩種溫度品系主要解毒酶活性的影響本實(shí)驗(yàn)研究了阿維菌素亞致死劑量處理,對(duì)巴氏新小綏螨兩種溫度品系體內(nèi)GSTs、MFOs和Car Es等解毒酶活性影響的劑量效應(yīng)和時(shí)間效應(yīng)。結(jié)果表明阿維菌素亞致死劑量對(duì)巴氏新小綏螨兩種溫度品系GSTs活性均有一定的抑制作用,存在一定的劑量效應(yīng)和時(shí)間效應(yīng),即劑量越大,相同濃度處理時(shí)間越長,抑制作用也越強(qiáng),且對(duì)耐高溫品系的抑制作用較常溫品系更強(qiáng)。阿維菌素亞致死劑量對(duì)巴氏新小綏螨兩種溫度品系MFOs和Car Es活性均有一定的誘導(dǎo)作用。具體表現(xiàn)為在阿維菌素濃度相對(duì)較低時(shí),其對(duì)巴氏新小綏螨兩種溫度品系MFOs和Car Es的誘導(dǎo)作用較強(qiáng),活性升高,以抵御外界不良環(huán)境,而后隨著濃度增大,誘導(dǎo)作用減弱;阿維菌素LC30處理時(shí)間越長,對(duì)耐高溫品系MFOs和Car Es的誘導(dǎo)作用越強(qiáng),但對(duì)常溫品系MFOs和Car Es的誘導(dǎo)作用則會(huì)有所減弱。4.巴氏新小綏螨兩種溫度品系阿維菌素脅迫下解毒代謝基因的表達(dá)模式分析本實(shí)驗(yàn)從巴氏新小綏螨轉(zhuǎn)錄組數(shù)據(jù)中找出獲得注釋的三種解毒代謝基因,對(duì)其進(jìn)行系統(tǒng)進(jìn)化分析后,利用內(nèi)參基因(Beta-actin)作為參照,使用q PCR技術(shù)對(duì)解毒代謝基因進(jìn)行驗(yàn)證,研究分析巴氏新小綏螨常溫品系和耐高溫品系10個(gè)解毒代謝基因(4個(gè)GSTs基因、4個(gè)MFOs基因以及2個(gè)Car Es基因)對(duì)阿維菌素脅迫響應(yīng)的濃度效應(yīng)和時(shí)間效應(yīng),明確了這10個(gè)解毒代謝基因在阿維菌素誘導(dǎo)后的m RNA表達(dá)量的變化趨勢(shì),以期為培育抗藥品系的巴氏新小綏螨提供參考。結(jié)果表明巴氏新小綏螨常溫品系:GST1、GST2、CYP3和CYP4的相對(duì)表達(dá)量在不同濃度以及LC30處理不同時(shí)間后均下調(diào),但下調(diào)幅度有所差別;GST3在阿維菌素劑量較小時(shí)其相對(duì)表達(dá)量有所下調(diào),劑量較大時(shí)其相對(duì)表達(dá)量上調(diào),其在LC30處理不同時(shí)間后相對(duì)表達(dá)量均上調(diào),但上調(diào)的幅度隨著時(shí)間的延長會(huì)有所變化;GST4在阿維菌素各劑量處理后,其相對(duì)表達(dá)量均上調(diào),但上調(diào)的幅度有所差別,其在阿維菌素LC30處理6h和24h后相對(duì)表達(dá)量均上調(diào),在LC30處理12h后相對(duì)表達(dá)量下調(diào);CYP1的相對(duì)表達(dá)量隨著阿維菌素劑量的增大而增大,其在LC30處理6h和12h后相對(duì)表達(dá)量均下調(diào),但在處理24h后,相對(duì)表達(dá)量卻表現(xiàn)上調(diào);CYP2的相對(duì)表達(dá)量在LC10、LC30處理組有所下調(diào),在LC50處理組卻表現(xiàn)上調(diào),其在LC30處理不同時(shí)間后相對(duì)表達(dá)量全部下調(diào);Car E1和Car E2的相對(duì)表達(dá)量均在阿維菌素劑量較小時(shí)表現(xiàn)上調(diào),隨著劑量加大,其上調(diào)幅度減小或甚至下調(diào),LC30脅迫初期(6h)有所上調(diào),隨著脅迫時(shí)間延長,其相對(duì)表達(dá)量下調(diào),但隨后又有所回升。表明GST1、GST2、CYP3和CYP4可能不參與巴氏新小綏螨常溫品系對(duì)阿維菌素的解毒代謝;而GST3、GST4、CYP1、CYP2、Car E1和Car E2可能都有參與巴氏新小綏螨常溫品系對(duì)阿維菌素的解毒代謝,因而它們也可能參與巴氏新小綏螨常溫品系對(duì)阿維菌素的抗性形成。巴氏新小綏螨耐高溫品系:GST1、GST2的相對(duì)表達(dá)量在阿維菌素劑量較小時(shí)表現(xiàn)上調(diào),但幅度有所差別,劑量達(dá)到LC50時(shí)卻表現(xiàn)下調(diào),其在阿維菌素LC30處理不同時(shí)間后相對(duì)表達(dá)量均上調(diào),但幅度有所差別,于12h處理組達(dá)到最大;GST3的相對(duì)表達(dá)量在阿維菌素劑量較小時(shí)表現(xiàn)上調(diào),但幅度有所差別,劑量達(dá)到LC50時(shí)卻下調(diào)了,其在阿維菌素LC30處理6h和12h后有所下調(diào),處理24h后卻上調(diào)了;GST4在阿維菌素各劑量處理后,其相對(duì)表達(dá)量均上調(diào),且隨著阿維菌素劑量的增大,其上調(diào)幅度也變大,其在阿維菌素LC30處理不同時(shí)間后相對(duì)表達(dá)量均上調(diào),但幅度有所差別,于12h處理組達(dá)到最大;CYP1在阿維菌素各劑量處理后,其相對(duì)表達(dá)量均上調(diào),隨著劑量增大上調(diào)幅度有所下降,其在阿維菌素LC30處理不同時(shí)間后相對(duì)表達(dá)量均上調(diào),但均無顯著性差異;CYP2在阿維菌素LC10處理后,其相對(duì)表達(dá)量上調(diào),隨著劑量增大,相對(duì)表達(dá)量卻下調(diào)了,其在阿維菌素LC30處理6h后上調(diào)至對(duì)照的7.27倍,隨著處理時(shí)間的延長,其相對(duì)表達(dá)量的上調(diào)幅度減小甚至下調(diào);CYP3、CYP4、Car E1和Car E2的相對(duì)表達(dá)量均在阿維菌素劑量較小時(shí)表現(xiàn)上調(diào),隨著劑量加大,其上調(diào)幅度減小或甚至下調(diào),其在阿維菌素LC30處理不同時(shí)間后均上調(diào),但上調(diào)幅度先增大后減小,于6h處理組達(dá)到最大。表明GST1、GST2、GST3、GST4、CYP1、CYP2、CYP3、CYP4、Car E1和Car E2可能都有參與巴氏新小綏螨耐高溫品系對(duì)阿維菌素的解毒代謝,因而它們也可能參與巴氏新小綏螨耐高溫品系對(duì)阿維菌素的抗性形成。通過以上幾個(gè)方面的研究,不僅可為巴氏新小綏螨常溫品系和耐高溫品系的田間釋放提供一定的參考,還能判斷解毒酶系以及某一解毒代謝基因在巴氏新小綏螨兩種溫度品系體內(nèi)所起的作用,為巴氏新小綏螨抗阿維菌素品系的篩選提供參考依據(jù)。
[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.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S476.2

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