本文選題：華山松大小蠹　切入點：秦嶺細粘束孢　出處：《西北農(nóng)林科技大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：細胞色素P450酶作為昆蟲重要的解毒酶系,不僅與昆蟲信息素和激素合成密切相關(guān),而且在降解寄主次生代謝毒素和殺蟲劑等方面發(fā)揮著重要的作用。小蠹蟲在入侵寄主樹木,以及在寄主樹木韌皮部和木質(zhì)部組織內(nèi)生存繁殖的整個生命過程中必須具備克服寄主樹木原生性和誘導(dǎo)性抗性,降解寄主樹木樹脂(萜類物質(zhì))毒害的能力。本研究選擇在我國秦嶺巴山林區(qū)持續(xù)發(fā)生的重大森林害蟲——華山松大小蠹(Dendroctonus armandi)為研究對象,旨在通過對華山松大小蠹細胞色素P450基因系統(tǒng)發(fā)育,華山松大小蠹細胞色素P450基因在不同發(fā)育階段與組織部位表達的差異與分布,華山松大小蠹成蟲對華山松揮發(fā)性物質(zhì)的耐受性,以及華山松大小蠹和共生真菌秦嶺細粘束孢(Leptographium qinlingensis)P450基因?qū)︔祁愇镔|(zhì)的響應(yīng)模式等研究,揭示華山松大小蠹P450基因的多樣性,華山松大小蠹克服寄主華山松多元抗性相關(guān)的P450基因類群,華山松大小蠹和共生真菌P450基因在克服寄主華山松抗性和降解寄主華山松萜類物質(zhì)毒害中的作用等重大科學(xué)問題,取得以下研究結(jié)果:1.華山松大小蠹成蟲對寄主華山松5種單萜類化合物((±)-α-Pinene、(-)-α-Pinene、(-)-β-Pinene、(+)-3-Carene、(±)-Limonene)和華山松萜類提取物耐受性存在顯著差異,寄主華山松萜類物質(zhì)對華山松大小蠹具有明顯的抗性和致死閾值,當(dāng)萜類物質(zhì)濃度超過閾值(600 ppm)死亡率迅速上升。2.華山松大小蠹至少具有64條P450基因,且在家族上存在多樣性。在進化分枝CYP3中存在小蠹蟲特異性P450基因,它們的功能與華山松大小蠹適應(yīng)特殊生境、克服寄主華山松多元抗性和華山松大小蠹特有的行為與生活史密切相關(guān)。3.華山松大小蠹43條P450基因在不同發(fā)育階段表達的反映了華山松大小蠹P450基因時間表達的差異性與功能的多樣性,其中在華山松大小蠹幼蟲向蛹和成蟲的發(fā)育過程中P450基因表達差異顯著,成蟲階段隨著羽化、擴散和入侵,華山松大小蠹成蟲CYP3、CYP4進化分枝的P450基因表達差異顯著,這說明在華山松大小蠹幼蟲、蛹和成蟲的整個發(fā)育過程中有多種P450酶的參與,華山松大小蠹P450基因在時間和發(fā)育階段的表達差異性與華山松大小蠹不同時間和發(fā)育階段所需要克服寄主華山松萜烯類物質(zhì)抗性的差異具有密切的關(guān)系,而且與華山松大小蠹寄主選擇有關(guān)。4.華山松大小蠹43條P450基因在雌雄成蟲的觸角、消化道和生殖器中表達差異顯著,且具有性別差異性,這說明華山松大小蠹P450基因在性別或組織中具有功能的特異性。華山松大小蠹消化道特異表達的P450基因與降低華山松樹脂毒害密切相關(guān),觸角中特異性表達的P450基因與降解氣味分子和寄主選擇密切相關(guān)。5.華山松大小蠹關(guān)鍵P450基因和秦嶺細粘束孢(Leptographium qinlingensis)P450基因?qū)闹魅A山松萜類物質(zhì)具有特殊的響應(yīng)模式,華山松大小蠹和共生真菌通過調(diào)控自身P450酶的表達水平,降低寄主華山松樹脂等對華山松大小蠹的毒害。6.華山松大小蠹共生真菌秦嶺細粘束孢CYP51F1基因具有典型的14-α-脫甲基酶蛋白活性位點和膜錨定結(jié)構(gòu)域,對華山松萜烯類物質(zhì)中的三萜(甾醇類化合物)具有顯著的降解作用,從而構(gòu)成華山松大小蠹和共生真菌協(xié)同作用克服寄主樹木抗性的機制。
[Abstract]:Cytochrome P450 enzyme as an importantdetoxification enzyme of insects, and insects not only pheromones and hormones are closely related, but also play an important role in the degradation of primary and secondary students send metabolic toxins and pesticides etc. bristletail in the invasion of host trees, and the survival of the entire life in the process of reproduction must have overcome original and host trees induced resistance in the host tree phloem and xylem tissues, degradation of host tree resin (terpenoids) poison ability. This study continues to occur in China's Qinling Mountains Bashan forests of the major forest pests in Huashan pine beetle (Dendroctonus armandi) as the research object, through the development of Huashan pine beetle cells cytochrome P450 gene system, differential expression of Dendroctonus cytochrome P450 gene in different developmental stages and tissues and the distribution of Huashan pine, Huashan pine beetle into Tolerance to insect volatile substances of Huashan pine, and Huashan pine beetle and symbiotic fungi in Qinling Mountains fine beam (Leptographium qinlingensis) spore of P450 gene on the terpenoid response pattern, diversity revealed Huashan Dendroctonus P450 gene, Huashan pine beetle overcome host Huashan P450 gene related to resistance to multiple groups of pine the Huashan pine beetle and symbiotic fungus P450 gene in host resistance and degradation of Huashan pine to overcome the host role of Huashan pinene toxicity in some important scientific problems, and achieved the following results: 1. Huashan pine beetle adults to host Huashan pine 5 monoterpenes ((+) - alpha -Pinene. (-) -Pinene (-) - alpha, beta -Pinene (+) -3-Carene (+) -Limonene) and Huashan pine terpenoids extract tolerance are significantly different, the host Huashan pine terpenes of Huashan pine beetle has obvious resistance and lethal threshold When the value of the terpenoid concentration exceeds the threshold (600 ppm) mortality rose rapidly.2. Huashan pine beetle has at least 64 P450 genes, and there exists diversity in the family. There are bristletail specific P450 gene in clade CYP3, their functions and the Huashan pine beetle adapt to special habitat, overcome host Huashan pine and Huashan pine beetle resistance multiple unique behavior and life history of.3. is closely related to the Huashan pine beetle of 43 P450 genes in different developmental stages of expression reflects the diversity of function and expression of P450 gene of Huashan pine beetle time, the pine beetle larvae to pupae in Huashan P450 development process the adults and the gene expression in the adult stage had significant difference, with the emergence, proliferation and invasion, CYP3 size of Huashan pine beetle, the differential expression of P450 gene CYP4 evolutionary branching significantly, which shows in Huashan pine beetle larvae There are a variety of insects, the P450 enzyme involved in the whole developmental process of pupae and adults in Huashan, difference of Dendroctonus P450 gene in time and developmental expression difference and Huashan pine beetle in different time and stage of development to overcome host Huashan pine terpenes resistance has close relationship with Huashan, and the size of pine choose the.4. Huashan host beetle Dendroctonus P450 43 gene in male and female adults, differential expression of digestive tract and genital significantly, and have gender differences, indicating the specificity of Huashan pine beetle has the function of P450 gene in sex or tissues. The P450 gene of Huashan pine beetle digestive tract specific expression closely associated with the reduction of Huashan pine resin toxicity, specific expression of P450 gene in the antennae and degradation of odorants and host selection of.5. is closely related to the Huashan pine beetle key gene and P450 in Qinling Mountains Fine beam (Leptographium qinlingensis) from P450 gene has a special response pattern to host Huashan pine terpenes, Huashan pine beetle and its symbiotic fungi by regulating the expression levels of the P450 enzyme, reduce the toxicity of.6. beetle host Huashan pine resin on Huashan pine and Huashan pine beetle symbiotic fungi in Qinling Mountains fine stick bundle spore the CYP51F1 gene has typical 14- alpha demethylase protein active site and membrane anchoring domain of Huashan pine, three terpene terpenes in (sterols) has significant degradation effect, which constitute the Huashan pine beetle and symbiotic fungus synergy mechanism to overcome host tree resistance.

【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S763.38
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本文編號：1600006