本文選題：亞洲玉米螟 + 絲素輕鏈基因��； 參考：《揚(yáng)州大學(xué)》2015年碩士論文

【摘要】：亞洲玉米螟(Ostrinia furnacalis)是一種分布廣泛且多食性的害蟲(chóng),對(duì)我國(guó)的玉米、棉花危害較大。為研究亞洲玉米螟的吐絲機(jī)理,本文克隆了亞洲玉米螟的絲素輕鏈基因片段且進(jìn)行了全長(zhǎng)拼接,并研究了在其不同發(fā)育階段、不同溫度、不同寄主條件下該基因的相對(duì)表達(dá)量；同時(shí),通過(guò)飼喂法對(duì)亞洲玉米螟絲素輕鏈基因進(jìn)行了RNA干擾�，F(xiàn)將研究結(jié)果報(bào)告如下：1、采用RACE技術(shù)克隆到亞洲玉米螟絲素輕鏈基因的cDNA全長(zhǎng)(已提交GeneBank: KF669645)。該基因全長(zhǎng)1098bp,自39bp至830bp為其開(kāi)放閱讀框,起始密碼子ATG,終止密碼子TAA,一共編碼263個(gè)氨基酸,相對(duì)分子量為92.76kDa,等電點(diǎn)pI為4.99。通過(guò)同源性比對(duì),亞洲玉米螟絲素輕鏈基因與棉大卷葉螟(Sylepta derogata Fabricius)同源性最高,為75%,與外米綴蛾(Corcyra cephalonica)的同源性為69%；與大蠟螟(Galleria mellonella)的同源性為68%。2、運(yùn)用實(shí)時(shí)熒光定量PCR,檢測(cè)了在不同發(fā)育階段、不同溫度條件、喂食不同寄主后亞洲玉米螟絲素輕鏈基因的相對(duì)表達(dá)量。結(jié)果表明,絲素輕鏈基因在5齡幼蟲(chóng)時(shí)相對(duì)表達(dá)量最高,其次是2齡幼蟲(chóng)與3齡幼蟲(chóng)；當(dāng)幼蟲(chóng)進(jìn)入預(yù)蛹后,該基因相對(duì)表達(dá)量開(kāi)始降低,至成蟲(chóng)時(shí),相對(duì)表達(dá)量幾乎為零；研究了22℃,25℃,28℃,31℃,34℃條件下亞洲玉米螟絲素輕鏈基因的相對(duì)表達(dá)量,發(fā)現(xiàn)34℃時(shí)相對(duì)表達(dá)量最高；在棉花、玉米、酸模葉蓼三種寄主間,取食棉花的亞洲玉米螟絲素輕鏈基因的表達(dá)量最高,其次分別是玉米、酸模葉蓼。文章對(duì)處于逆境條件下玉米螟絲素輕鏈基因的相對(duì)表達(dá)量較高進(jìn)行了討論。3、通過(guò)飼喂法,將表達(dá)目標(biāo)基因dsRNA的菌液飼喂亞洲玉米螟幼蟲(chóng),檢測(cè)沉默效果。結(jié)果發(fā)現(xiàn),相對(duì)于喂食添加了清水的常規(guī)飼料的亞洲玉米螟幼蟲(chóng)(CK),喂食添加了表達(dá)dsRNA菌液飼料的亞洲玉米螟的絲素輕鏈基因表達(dá)量分別下調(diào)了34.81%、35.05%。經(jīng)過(guò)RNA干擾后,相對(duì)于對(duì)照,亞洲玉米螟的幼蟲(chóng)發(fā)育歷期縮短0.99d、蛹重下降8.53%、單雌產(chǎn)卵量減少34.86%。這表明絲素輕鏈基因表達(dá)量的降低與亞洲玉米螟的生長(zhǎng)發(fā)育是有關(guān)聯(lián)的。
[Abstract]:Asian corn borer (Ostrinia furnacalis) is a widely distributed and multifood pest, which is harmful to maize and cotton in China. In order to study the silking mechanism of Asian corn borer, the silk light chain gene fragment of Asian corn borer was cloned and spliced, and the relative expression amount of the gene was studied at different development stages, different temperatures and different host conditions. At the same time, RNA interference of Asian corn borer fibroin light chain gene was carried out by feeding method. The results are reported as follows: 1. The full-length cDNA of Asian corn borer fibroin light chain gene was cloned by race technique (GeneBank: KF669645). The total length of the gene is 1098bp. the open reading frame is from 39bp to 830bp, the start codon is ATG, and the stop codon is TAA. It encodes 263 amino acids, the relative molecular weight is 92.76 kDaand the isoelectric point Pi is 4.99. By homology comparison, Asian corn borer silk fibroin light chain gene has the highest homology with Sylepta derogata Fabricius, which is 75% and 69% with Corcyra cephalonica. The homology with Galleria mellonella was 68.2. The relative expression of silk fibroin light chain gene of Asian corn borer at different development stage and different temperature was detected by real-time fluorescence quantitative PCR. The results showed that the relative expression of silk fibroin light chain gene was the highest in the 5th instar larva, followed by the 2nd instar larva and the 3rd instar larva, and the relative expression level of the fibroin light chain gene began to decrease when the larva entered the prepupae, and almost zero at the adult stage. The relative expression of fibroin light chain gene of Asian corn borer was studied at 22 鈩，

本文編號(hào)：2061654