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  本文關(guān)鍵詞：小菜蛾保幼激素受體基因的分子特性及生殖調(diào)控作用研究　出處：《福建農(nóng)林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 小菜蛾 保幼激素 保幼激素受體 生殖調(diào)控 害蟲防治

【摘要】：小菜蛾P(guān)lutella xyPostella(L.)(鱗翅目菜蛾科Lepidoptera,Plutellidae)是危害十字花科蔬菜的世界性害蟲,生殖能力強是其成為全球分布最廣的鱗翅目害蟲的重要因素之一。保幼激素(juvenile hormone,JH)是參與昆蟲生殖調(diào)控的重要物質(zhì),在昆蟲卵子發(fā)生中起著至關(guān)重要的作用,但對其分子機制的了解仍然有限,尤其是在非模式昆蟲中。保幼激素受體(methoprene-tolerant,Met)作為JH的關(guān)鍵受體之一,對JH發(fā)揮功能效應(yīng)起著重要調(diào)節(jié)作用,但具有明顯的物種特異性。本研究圍繞生殖調(diào)控決定昆蟲生殖潛能這一科學(xué)問題,以激素調(diào)控生殖為核心,采用生物化學(xué)與分子生物學(xué)、解剖形態(tài)學(xué)與實驗生物學(xué)等方法,從JH的作用受體方面開展研究,重點研究:小菜蛾Met基因的分子結(jié)構(gòu)特征與進化關(guān)系,小菜蛾Met基因在不同齡期與不同組織的表達模式,以及Met基因在小菜蛾生殖調(diào)控中的作用。主要結(jié)果如下:PxMet-1與PxMet-2開放閱讀框(ORF)全長分別為1575和2100bp,預(yù)計編碼524和699個氨基酸,理論分子質(zhì)量為60.5和70.7 kDa,預(yù)測等電點為6.73和5.50。PxMet-1和PxMet-2都具有4個保守結(jié)構(gòu)域,分別是Helix-Loop-Helix結(jié)構(gòu)域(bHLH)以及兩個PAS保守結(jié)構(gòu)域及一個PAC保守基序。氨基酸序列同源比對分析結(jié)構(gòu)顯示,PxMet-1與PxMet-2與鱗翅目昆蟲中的家蠶、棉鈴蟲、冬尺蠖蛾的相似性均大于50%。進化樹分析結(jié)果顯示,小菜蛾P(guān)xMet-1,PxMet-2聚為不同的兩支,說明PxMet-1,PxMet-可能具有不同的生物學(xué)功能以及進化模式,但兩者均與鱗翅目昆蟲聚在一起,且分別與家蠶的PxMet-1與PxMet-2聚在一起,說明PxMet-與PxMet-2與鱗翅目昆蟲Mets可能發(fā)揮相同作。PxMet-1與PxMet-2在蛹期(1-3d)與雌成蟲期(0-72h)均有表達,PxMet-1在蛹期的表達量無明顯差異,但均顯著高于雌成蟲期(0-48h);而PxMet-2在雌成蟲期(0-48 h)的表達量呈先上升后下降的趨勢,在羽化后12 h出現(xiàn)表達高峰,且成蟲期(0-36h)的表達量顯著高于蛹期;PxMet-1與PxMet-2均在雌蟲羽化后72h出現(xiàn)一個明顯的表達升高,推測可能與小菜蛾生殖與壽命維持的能量權(quán)衡相關(guān)。PxMet-1與PxMet-2在脂肪體的表達量顯著高于其他組織(頭、腸道、卵巢、殘渣)。利用RNAi技術(shù)同時干擾PxMet-1和PxMet-2,結(jié)果顯示,在注射Met-l+Met-2 dsRNA24 h后,PxMet-1和PxMet-2的表達量均受到顯著抑制,但48 h與72 h時,干擾效應(yīng)明顯減弱,處理組與對照組表達量無顯著差異。干擾PxMet-1和PxMet-2后,小菜蛾卵巢發(fā)育受到明顯抑制,卵黃沉積減緩,成熟卵子數(shù)目減少,產(chǎn)卵量也隨之下降。本研究明確了小菜蛾Met的分子結(jié)構(gòu)特征、表達模式及基因功能,可為初步闡明JH調(diào)控小菜蛾生殖過程的分子信號途徑奠定基礎(chǔ)。在實踐上有助于篩選控制小菜蛾的潛在靶標(biāo),為有效防治和持續(xù)控制小菜蛾種群暴發(fā)與危害提供創(chuàng)新思路和途徑。
[Abstract]:Diamondback moth Plutella xyPostellaa L. (Lepidoptera, Lepidoptera). Plutellidaeis a worldwide pest that harms cruciferous vegetables. Strong reproductive ability is one of the most widely distributed Lepidoptera pests in the world, and juvenile hormone rejuvenile hormoney (JHH) is an important substance involved in insect reproductive regulation. It plays an important role in insect oogenesis, but its molecular mechanism is still limited. Especially in non-model insects, methoprene-tolerantantone is one of the key receptors of JH. JH plays an important role in regulating the function of JH, but it has obvious species-specificity. This study focuses on the scientific issue of reproductive regulation and determination of insect reproductive potential, with hormone regulation of reproduction as the core. By means of biochemical and molecular biology, anatomical morphology and experimental biology, the study was carried out from the aspect of JH acting receptor, with emphasis on the molecular structural characteristics and evolutionary relationship of the Met gene of Plutella xylostella (Plutella xylostella). Expression patterns of Met gene in different age and tissue of Plutella xylostella. And the role of Met gene in the reproductive regulation of Plutella xylostella. The main results were as follows: 1 575 BP and 2100bp of PxMet-2 open reading frame (ORF). It is predicted to encode 524 and 699 amino acids with theoretical molecular weights of 60.5 and 70.7 kDa. The predicted isoelectric points were 6.73 and 5.50. PxMet-1 and PxMet-2 had four conserved domains. Helix-Loop-Helix domain, two PAS conserved domains and one PAC conserved motif. The similarity between PxMet-1 and PxMet-2 and Bombyx mori, Helicoverpa armigera and Inchworm in Lepidoptera was more than 50. The result of phylogenetic tree analysis showed that PxMet-1 of Plutella xylostella was similar. PxMet-2 was clustered into two different branches, indicating that PxMet-1 PxMet- may have different biological functions and evolutionary models, but both of them were clustered with Lepidoptera insects. And with silkworm PxMet-1 and PxMet-2. It is suggested that PxMet- and Mets of Lepidoptera and PxMet-2 may play the same role. PxMet-1 and PxMet-2 may play the same role in pupa stage (1-3 d) and female adult stage (0-72 h). All expressed. There was no significant difference in the expression of PxMet-1 in pupa stage, but it was significantly higher than that in female adult stage from 0 to 48 hours. However, the expression of PxMet-2 increased first and then decreased at 0-48 h after female adult stage, and reached a peak at 12 h after emergence. And the expression level of the adult stage was significantly higher than that of the pupa stage. Both PxMet-1 and PxMet-2 were significantly increased at 72 hours after emergence. It is speculated that the expression of .PxMet-1 and PxMet-2 in fat body is significantly higher than that in other tissues (head, intestine, ovary), which may be related to the energy tradeoff between reproduction and longevity of Plutella xylostella. Both PxMet-1 and PxMet-2 were interfered with by RNAi technique. The results showed that after Met-l Met-2 dsRNA24 h injection, both PxMet-1 and PxMet-2 were interfered with. The expression of PxMet-1 and PxMet-2 were significantly inhibited, but at 48 and 72 hours, the interference effect was significantly weakened. After interfering with PxMet-1 and PxMet-2, the ovary development of Plutella xylostella was significantly inhibited, the egg yolk deposition slowed down, and the number of mature eggs decreased. The molecular structure, expression pattern and gene function of Plutella xylostella Met were determined. It can lay a foundation for elucidating the molecular signal pathway of JH regulating the reproductive process of Plutella xylostella and is helpful to screen the potential targets for controlling Plutella xylostella in practice. To effectively control and sustainable control of diamondback moth population outbreak and harm to provide innovative ideas and ways.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S433.4

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