井冈霉素刺激褐飞虱生殖的蛋白质组学分析及关键基因FAS的功能研究

发布时间：2018-02-04 21:51

本文关键词： 褐飞虱井冈霉素蛋白质组学分析脂肪酸合酶 RNA干扰　出处：《扬州大学》2016年硕士论文　论文类型：学位论文

【摘要】：褐飞虱(同翅目、飞虱科)是典型的农药诱导再猖獗型水稻害虫。井冈霉素是防治水稻纹枯病的抗生素类杀菌剂,由于毒性低,残留低,成本低等特点,很容易被滥用。目前已发现井冈霉素喷雾处理和点滴处理均能刺激褐飞虱生殖,且喷雾处理的刺激作用大于点滴处理,但其刺激褐飞虱生殖的分子机制仍不清楚。本试验通过井冈霉素不同处理间褐飞虱卵巢的蛋白质组学分析,筛选出井冈霉素刺激褐飞虱生殖的关键蛋白,以阐明井冈霉素刺激褐飞虱生殖的分子机制,同时揭示喷雾处理刺激作用大于点滴处理的原因。1、井冈霉素不同处理间褐飞虱卵巢的蛋白质组学分析利用同重同位素相对与绝对定量技术,对井冈霉素各处理间褐飞虱卵巢进行蛋白质组学分析。喷雾井冈霉素(JGM-S) vs喷雾对照(S-control)处理组共鉴定出284个差异蛋白(142个上调,142个下调),点滴井冈霉素(JGM-T) vs点滴对照(T-control)处理组共鉴定出267个差异蛋白(130个上调,137个下调),喷雾井冈霉素(JGM-S) vs点滴井冈霉素(JGM-T)处理组共鉴定出114个差异蛋白(62个上调,52个下调)。通过pathway注释与富集分析,筛选出喷雾井冈霉素(JGM-S) vs喷雾对照(S-control)处理组与喷雾井冈霉素(JGM-S) vs点滴井冈霉素(JGM-T)处理组中都显著上调的脂肪酸合酶进一步研究。2、干扰脂肪酸合酶基因对褐飞虱生殖的影响利用饲喂法对井冈霉素喷雾处理(JGM-S)的褐飞虱进行RNA干扰,形成干扰处理JGM-S+dsFAS(脂肪酸合酶基因表达量下降了58%)。产卵结果表明,干扰脂肪酸合酶基因后,褐飞虱产卵量下降46%,产卵历期下降33%,雌虫寿命下降24%。生理生化测定结果表明,dsFAS处理显著降低了褐飞虱雌虫体重和卵巢中粗脂肪含量。JGM-S+dsFAS处理组褐飞虱雌虫卵巢中油酸、棕榈酸、亚油酸、硬脂酸、肉豆蔻酸,脂肪体中油酸、棕榈酸、肉豆蔻酸含量与对照(JGM-S)相比均显著下降。3、褐飞虱吸食井冈霉素后生理生化变化取食100ppm和20ppm的井冈霉素使褐飞虱脂肪酸合酶基因表达量分别提高了54%和56%,卵黄原蛋白基因表达量分别提高了133%和94%。选取20ppm井冈霉素进一步饲喂研究发现,褐飞虱雌虫可溶性糖含量提高了46%、胰岛素含量提高了17%、体重提高了10%,共生菌含量提高了38%。干扰脂肪酸合酶基因使褐飞虱雌虫体重显著下降,但对共生菌含量没有显著影响。
[Abstract]:Brown planthopper (Homoptera, planthopper) is a typical insecticide-induced re-rampant rice pest. Jinggangmycin is an antibiotic fungicide to control rice sheath blight, because of its low toxicity, low residue and low cost. It has been found that both spray treatment and drip treatment can stimulate the reproduction of brown planthopper, and the stimulative effect of spray treatment is greater than that of drip treatment. However, the molecular mechanism of the stimulation of brown planthopper reproduction is still unclear. Through proteomic analysis of the ovaries of brown planthopper in different treatments of Jinggangmycin, the key proteins of Jinggangmycin to stimulate the reproduction of brown planthopper were screened out. In order to elucidate the molecular mechanism of Jinggangmycin to stimulate the reproduction of brown planthopper, and to reveal the reason that spray treatment has more stimulative effect than that of drip treatment. Proteomics analysis of the ovaries of brown planthopper with different treatments of Jinggangmycin using the technique of iso-isotopic and absolute quantification. Proteomic analysis of the ovaries of brown planthopper treated with Jinggangmycin. JGM-S vs S-control). 284 differentially expressed proteins (142 up-regulated) were identified in the treatment group. A total of 267 differentially expressed proteins (130 up-regulated and 137 down-regulated) were identified in 142 down-regulated and infused Jinggangmycin (JGM-T) vs control (T-control) groups. A total of 114 differentially expressed proteins (62 up-regulated) were identified in the spray Jinggangmycin (JGM-S) vs drip Jinggangmycin (JGM-T) treatment group. 52 of them were down-regulated by pathway annotation and enrichment analysis. The jingangmycin (JGM-S) vs the spray control (S-control) treatment group and the spray Jinggangmycin (JGM-S) vs droplet Jinggangmycin (JGM-T) treatment group were screened out. Fatty acid synthase, which was significantly up-regulated in treatment group, was further studied. Effect of interfering Fatty Acid Synthase Gene on the Reproduction of Brown planthopper the RNA interference was carried out on the brown planthopper treated with jinggangmycin spray. The expression of fatty acid synthase gene in JGM-S dsFASs decreased by 58%. The results showed that the egg production of BPH decreased by 46% after interfering with the fatty acid synthase gene. The egg laying period decreased 33%, the female longevity decreased 24%. The physiological and biochemical results showed that. DsFAS treatment significantly decreased the body weight of female brown planthopper and the content of crude fat in ovary. JGM-S dsFAS treatment group showed oleic acid, palmitic acid, linoleic acid and stearic acid in ovary of female brown planthopper. The contents of myristic acid, oleic acid, palmitic acid and myristic acid in fat body were significantly lower than those in control group (JGM-S). Physiological and biochemical changes of Brown planthopper after ingesting Jinggangmycin 100 ppm and 20 ppm Jinggangmycin increased the expression of fatty acid synthase gene of brown planthopper by 54% and 56% respectively. The expression of vitellogenin gene was increased by 133% and 94ppm respectively. The results showed that the soluble sugar content of female brown planthopper increased by 46% by 20 ppm Jinggangmycin. Insulin content increased 17%, body weight increased 10%, symbiotic bacteria content increased 38%. Interference with fatty acid synthase gene significantly decreased body weight of female brown planthopper, but had no significant effect on symbiotic bacteria content.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.112.3

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