本文選題：酚氧化酶 + 槲皮素��； 參考：《山東農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：酚氧化酶(phenoloxidase,簡稱PO,EC.1.14.18.1)在昆蟲的生長發(fā)育和內(nèi)源性免疫過程中具有重要功能,如具有黑化或包被入侵的病原物、昆蟲傷口的愈合等功能。酚氧化酶可以作為害蟲控制中一個潛在的作用靶標,通過開發(fā)酚氧化酶抑制劑,對于害蟲綠色防控和環(huán)境生態(tài)安全提供新思路。本論文利用植物源的槲皮素和微生物農(nóng)藥球孢白僵菌處理小菜蛾,通過制備小菜蛾兩個酚氧化酶基因的多克隆抗體,研究不同處理對酚氧化酶蛋白表達水平的影響。酚氧化酶及其抑制劑也一直是國內(nèi)外科學(xué)家研究的熱點。本論文以小菜蛾為研究對象,在制備獲得小菜蛾P(guān)xPPO1和PxPPO2基因抗體的基礎(chǔ)之上,利用蛋白質(zhì)印跡技術(shù)(Western Blot)研究了槲皮素和球孢白僵菌(Beauveria bassiana)處理小菜蛾后,小菜蛾酚氧化酶蛋白翻譯表達水平的相對變化,從而來探討外來效應(yīng)物對酚氧化酶基因在翻譯水平上的影響。主要結(jié)果如下:1.成功制備獲得小菜蛾P(guān)xPPO1和PxPPO2基因的多克隆抗體。2.分別對內(nèi)參基因β-actin以及目的基因PxPPO1和PxPPO2進行了 Western Blot曝光飽和分析,根據(jù)上樣量與信號飽和點出現(xiàn)的時間,確定Western Blot試驗中上樣量為 10μg。3.采用微量點滴法,用不同濃度槲皮素處理小菜蛾3齡幼蟲,處理不同時間后,PxPPO1和PxPPO2基因翻譯相對表達量隨槲皮素濃度升高而明顯減少。4.采用浸漬法,用球孢白僵菌侵染小菜蛾3齡幼蟲,成功侵染不同時間后,PxPPO1和PxPPO2基因翻譯相對表達量在處理2h時比對照處理提高,在6h、12h、24h和48h時蛋白表達量均比對照處理減少。
[Abstract]:Phenoloxidase (POC. 1.14.18.1) plays an important role in the growth and development of insects and endogenous immune process, such as melanization or encapsulation of invading pathogens, wound healing of insects and so on. Phenoloxidase can be used as a potential target in pest control. Through the development of phenoloxidase inhibitors, it provides a new idea for pest green control and environmental ecological safety. In this paper, quercetin from plant and microbial pesticide Beauveria bassiana were used to treat diamondback moth. The polyclonal antibody of two phenoloxidase genes of Plutella xylostella was prepared to study the effect of different treatments on the expression of phenoloxidase protein. Phenoloxidase and its inhibitors have also been the focus of scientists at home and abroad. Based on the preparation of PxPPO1 and PxPPO2 gene antibodies from diamondback moth (Plutella xylostella), the effects of quercetin and Beauveria bassiana on diamondback moth (Plutella xylostella) were studied by Western blot. The relative changes of phenoloxidase protein translation and expression level in Plutella xylostella were used to explore the effect of foreign effectors on the translation level of phenoloxidase gene. The main results are as follows: 1. The polyclonal antibodies against PxPPO1 and PxPPO2 genes of Plutella xylostella were successfully prepared. The 尾 -actin gene and the target genes PxPPO1 and PxPPO2 were analyzed by Western Blot. According to the amount of sample and the time of signal saturation, it was determined that the amount of sample was 10 渭 g 路3 in Western Blot. The third instar larvae of Plutella xylostella were treated with quercetin at different concentrations by microdrop method. The relative expression of PxPPO1 and PxPPO2 genes decreased significantly with the increase of quercetin concentration. The third instar larvae of Plutella xylostella were infected with Beauveria bassiana by dipping method. The relative expression of PxPPO1 and PxPPO2 genes was increased at 2 h after successful infection, and decreased at 6 h, 12 h, 24 h and 48 h, respectively.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S433.4

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