本文選題：綠盲蝽 + 性信息素��； 參考：《華中農(nóng)業(yè)大學》2016年碩士論文

【摘要】：綠盲蝽(Apolygus lucorum)是我國危害棉花的盲蝽蟓優(yōu)勢種之一,目前主要依賴化學防治,研究綠盲蝽體內(nèi)重要基因的功能,對探尋綠色安全的防治措施具有重要意義。盲蝽性信息成分主要為短碳鏈的醛和酯組成,前人研究報道醇氧化酶(alcohol oxidase,AOX)、乙�；D移酶(acetyltransferase,ATF)和去飽和酶(desaturase,DES)是醛和酯類物質(zhì)的合成過程中的重要酶類,但它們在綠盲蝽中的功能并不清楚,因此本文開展了這三類基因在綠盲蝽體內(nèi)的功能研究。RNA注射干擾是本文基因功能研究的重要手段,我們首先研究了RNA注射干擾綠盲蝽的最佳注射體積,結果表明羽化第一天的綠盲蝽成蟲的最佳注射體積為100n L。本研究采用熒光定量PCR研究了8個ATF基因,2個AOX和1個DES基因在性信息素釋放高峰期和未成熟雌雄蟲臭腺中的表達差異,結果表明ATF基因NAA25在3齡綠盲蝽雌蟲臭腺中特異性表達的特異性高表達,因此NAA25可能與綠盲蝽性信息素合成相關。本研究采用顯微注射法對綠盲蝽的ATF基因NAA25、NAA20和NAA15進行了干擾,其中干擾NAA25后,性成熟綠盲蝽雌蟲體內(nèi)成熟卵子數(shù)量降低了87.1%(p0.005),干擾NAA20后,性成熟綠盲蝽雌蟲體內(nèi)成熟卵子數(shù)量降低了95.6%(p0.005),干擾NAA15后,性成熟綠盲蝽雌蟲體內(nèi)成熟卵子數(shù)量降低了98.8%(p0.005)。利用實時熒光定量檢測了這3個基因從羽化第一天到性成熟時在全蟲中的表達量,這3個基因均在羽化第一天和性成熟前一天高表達,由此我們推測這3個基因在卵巢發(fā)育中起著重要的功能。在本研究中,我們通過RACE技術獲得2個N-alpha-acetyltransferase(NAA)基因的全長。其中N-alpha-acetyltransferase 25(NAA25)屬于Nat B復合酶的非催化亞基,N-alpha-acetyltransferase 15(NAA15)屬于Nat A復合酶的輔助亞基。其中NAA25序列全長為2898bp,ORF(open reading frame)區(qū)域為2064bp,編碼687個氨基酸。NAA15序列全長為1103bp,ORF區(qū)域為759bp,編碼252個氨基酸。
[Abstract]:Apolygus lucorum (Apolygus lucorum) is one of the dominant species that harm cotton in China. At present, it is very important to study the function of important genes in the plant by chemical control. It is of great significance to explore the control measures of green safety. The sex information components of stink bug mainly consist of aldehydes and esters of short carbon chain. Previous studies have reported that alcohol oxidase, acetyltransferase (ATF) and desaturase (DES) are important enzymes in the synthesis of aldehydes and esters, and some previous studies have reported that alcohol oxidase, acetyltransferase (ATF) and desaturase (DES) are important enzymes in the synthesis of aldehydes and esters. However, the function of these three genes in green bug is not clear. RNA injection interference is an important method to study the gene function in this paper. We first studied the optimal injection volume of RNA to interfere with the green bug. The results showed that the best injection volume of the adult was 100nL on the first day of Eclosion. In this study, the expression of 8 ATF genes, 2 AOX genes and 1 DES gene were studied by fluorescence quantitative PCR in the osmidine glands of male and female females during the peak period of sex pheromone release. The results showed that the specific expression of ATF gene NAA25 was highly specific in the stink glands of the third instar female bugs, so NAA25 might be related to the synthesis of sex pheromones. In this study, the microinjection method was used to interfere with the ATF gene NAA25, NAA20 and NAA15. After interfering with NAA25, the number of mature eggs decreased by 87.1% (P 0.005) and NAA20 (P < 0.05), respectively. The number of mature eggs decreased by 95.6% (p 0.005), and the number of mature eggs decreased by 98.8% (P 0.005) after interfering with NAA15. The expression of these three genes in the whole insect from the first day of emergence to the first day of sexual maturity was detected by real-time fluorescence quantitative analysis. The three genes were highly expressed on the first day of emergence and the day before sexual maturation. Therefore, we speculate that these three genes play an important role in ovarian development. In this study, we obtained two full length N-alpha-acetyltransferase NAA genes by RACE technique. Among them, N-alpha-acetyltransferase 25 (NAA25) belongs to the non-catalytic subunit N-alpha-acetyltransferase 15 (NAA15) of Nat B complex enzyme. It belongs to the auxiliary subunit of Nat A complex enzyme. The total length of NAA25 sequence is 2898bpSORF open reading frame region is 2064bp. the total length of NAA25 sequence encoding 67-amino acid. NAA15 sequence is 1103bpHORF region is 759bp, encoding 252amino acid.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S433

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