本文選題：斜紋夜蛾　切入點：性信息素　出處：《中國計量學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：斜紋夜蛾(Spodoptera litura)是危害性嚴(yán)重的鱗翅目害蟲,它具有進(jìn)化完善的性信息素通訊系統(tǒng)。斜紋夜蛾雌蟲在信息素腺體釋放性信息素,并通過它來吸引雄蟲進(jìn)而引發(fā)后續(xù)的交配行為。斜紋夜蛾性信息素是4種14碳直鏈不飽和脂肪酸酯類衍生物的混合物,它們的合成經(jīng)歷了軟脂酸合成、碳鏈縮短反應(yīng)、脫飽和作用和官能基團(tuán)的修飾這一系列過程。在該途徑中,多種酶參與其中并催化不同的生化反應(yīng)。本研究參照已有的斜紋夜蛾轉(zhuǎn)錄組數(shù)據(jù)并結(jié)合斜紋夜蛾的性信息素合成途徑,我們對其中的一系列相關(guān)基因進(jìn)行了篩選,并將之與NCBI數(shù)據(jù)庫中其他物種的性信息素合成途徑基因進(jìn)行比對,最終篩選到包括脂肪酸合成酶系、β氧化酶系、脫飽和酶、脂肪酰輔酶A還原酶和乙酰輔酶A轉(zhuǎn)移酶5大類共90個基因。為了進(jìn)一步確認(rèn)它們的表達(dá)特征,我們分別以雌、雄斜紋夜蛾為模板通過PCR進(jìn)行了半定量篩選。結(jié)果顯示,大多數(shù)基因在雌、雄斜紋夜蛾腹部表達(dá)沒有顯著差異;然而,我們發(fā)現(xiàn)少部分基因在雌性個體中特異性表達(dá);另外還有一部分基因在雌雄個體中的表達(dá)水平有顯著差異,因此我們通過熒光定量PCR進(jìn)一步檢測它們的表達(dá)水平并計算出表達(dá)差異的倍數(shù),結(jié)果表明這部分基因在雌蟲中的表達(dá)水平均高于雄蟲3-10倍;我們針對雌蟲特異性表達(dá)基因,研究了它們在雌蟲信息素腺體和腹部其他組織中的表達(dá),結(jié)果表明這些基因絕大多數(shù)在信息素腺體中的表達(dá)水平顯著高于腹部其他組織(p0.05),只有極少數(shù)基因在腹部其他組織中的表達(dá)量高于性信息素腺體。此外,我們選擇了上述篩選到的屬于不同類型(脂肪酸合成酶、脫飽和酶、脂酰輔酶A還原酶),且具有不同的表達(dá)特征的9個基因作進(jìn)一步的研究�？寺×诉@9個基因并合成了雙鏈RNA,然后將雙鏈RNA注射到斜紋夜蛾雌蟲進(jìn)行基因沉默,通過熒光定量PCR檢測基因沉默效果,根據(jù)GC-MS分析基因沉默后的雌蟲性信息素組分的變化。結(jié)果表明注射雙鏈RNA后對應(yīng)的基因表達(dá)量降低,并且對Des3,Des5和FAR17沉默后發(fā)現(xiàn)雌蟲產(chǎn)生的4中性信息素組分相對比例發(fā)生了變化。最后我們研究了雌蟲特異性基因在不同地理區(qū)系的表達(dá)差異,結(jié)果表明大多數(shù)基因在不同地域表達(dá)有顯著差異,而有的基因在不同地域表達(dá)水平相對穩(wěn)定。研究斜紋夜蛾性信息素合成途徑相關(guān)基因的功能和不同地理區(qū)系表達(dá)特征,有助于我們理解性信息素的種內(nèi)變異并推測它的進(jìn)化方向,此外為生物合成性信息擾亂蛾類交配提供理論依據(jù),也為在田間應(yīng)用性信息素防治害蟲提供了重要的支持。
[Abstract]:Spodoptera litura (Spodoptera litura) is a serious pest of Lepidoptera, which has a well-evolved sex pheromone communication system. Females of Spodoptera litura release sex pheromone in pheromone glands. The sex pheromone of Spodoptera litura is a mixture of four 14 carbon straight chain unsaturated fatty acid esters, which undergo palmitic acid synthesis and carbon chain shortening. Desaturation and modification of functional groups. In this pathway, According to the transcriptional data of Spodoptera litura and the sex pheromone synthesis pathway of Spodoptera litura, a series of related genes were screened. The genes of sex pheromone biosynthesis pathway of other species in NCBI database were compared, and finally selected to include fatty acid synthase, 尾 oxidase, desaturase. There are 90 genes in 5 categories of fatty acyl coenzyme A reductase and acetyl coenzyme A transferase. In order to further confirm their expression characteristics, we used female and male Spodoptera litura as templates to carry out semi-quantitative screening by PCR. There was no significant difference in the abdominal expression of most genes between female and male Spodoptera litura, however, we found that a few genes were specifically expressed in female individuals, and there were also significant differences in the expression levels of some genes in female and male individuals. Therefore, we further detected their expression level by fluorescence quantitative PCR and calculated the multiple of the difference in expression. The results showed that the expression level of these genes in females was 3-10 times higher than that in males, and we expressed genes specifically for females. Having studied their expression in female pheromone glands and other abdominal tissues, The results showed that the expression level of these genes was significantly higher in pheromone glands than in other abdominal tissues, and only a few genes expressed more in other abdominal tissues than in sex pheromones. We selected different types of enzymes (fatty acid synthase, desaturase, desaturase). Nine genes with different expression characteristics were further studied. The nine genes were cloned and synthesized double-stranded RNAs. Then double-stranded RNA was injected into female Spodoptera litura for gene silencing. The effect of gene silencing was detected by fluorescence quantitative PCR, and the changes of sex pheromone components of female worm after gene silencing were analyzed according to GC-MS. The results showed that the corresponding gene expression decreased after injection of double-stranded RNA. The relative proportion of 4 neutral pheromones produced by female worms was changed after the silencing of Des3, Des5 and FAR17. Finally, we studied the difference of female specific gene expression in different geographical regions. The results showed that most genes expressed differently in different regions, while some genes were relatively stable in different regions. The function of genes related to sex pheromone biosynthesis pathway of Spodoptera litura and the expression characteristics of different geographical regions were studied. It is helpful for us to understand the intraspecific variation of sex pheromone and to speculate its evolutionary direction. In addition, it provides a theoretical basis for biosynthetic information to disrupt the mating of moths, and also provides important support for the application of pheromones to pest control in the field.
【學(xué)位授予單位】：中國計量學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S433.4

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 殷坤山;洪北邊;洪子華;;茶尺蠖性信息素腺體部位的研究[J];中國茶葉;1990年05期

2 黃昌本,吳松筠,符文俊;棉褐帶卷葉蛾安徽種群不同日齡雌蛾性信息素的個體變異[J];華東昆蟲學(xué)報;2000年01期

3 王世偉;激素和性信息素在防治害蟲上的應(yīng)用[J];農(nóng)村實用技術(shù);2004年01期

4 薛勇;;激素和性信息素在防治害蟲上的應(yīng)用技術(shù)[J];農(nóng)藥市場信息;2005年14期

5 盧行尚;盧純高;;性信息素誘殺技術(shù)在隆安縣無公害生產(chǎn)上的應(yīng)用[J];廣西植保;2009年S1期

6 胡玉偉;管楚雄;許漢亮;林明江;安玉興;張志林;;害蟲性信息素種群監(jiān)測和大量誘殺技術(shù)的應(yīng)用[J];廣東農(nóng)業(yè)科學(xué);2013年16期

7 程妼青,肖錦澄,黃■廷,陳登隆,李健全,何炎聲,黃尚仁,羅慶昌,楊朝明,楊贊熹;中國玉米螟蟲的性信息素[J];自然雜志;1980年04期

8 楊堅;;昆蟲的性信息素[J];科技簡報;1980年05期

9 趙博光,王曉蕓,黃悅悅;大袋蛾(Clania varigata Snell.)雌蟲性信息素分泌腺體及其釋放機(jī)制的研究——Ⅱ.性信息素的室內(nèi)定量生測[J];南京林業(yè)大學(xué)學(xué)報(自然科學(xué)版);1982年04期

10 顧增光;;應(yīng)用性信息素防治棉花紅鈴蟲[J];浙江科技簡報;1982年06期

相關(guān)會議論文 前10條

1 張濤;楊代斌;李永紅;袁會珠;寧君;梅向東;;盲蝽科害蟲性信息素研究進(jìn)展[A];植物保護(hù)科技創(chuàng)新與發(fā)展——中國植物保護(hù)學(xué)會2008年學(xué)術(shù)年會論文集[C];2008年

2 孔杰;劉s，

本文編號：1588001