本文關(guān)鍵詞： 棉花 病毒介導的基因沉默(VIGS) 多酚氧化酶基因(PPO) 抗蟲性 基因表達　出處：《農(nóng)業(yè)生物技術(shù)學報》2017年05期 　論文類型：期刊論文

【摘要】：多酚氧化酶(polyphenol oxidase,PPO)是一類含銅的氧化還原酶,能夠抵御植食性昆蟲或病原菌的危害。為了研究棉花(Gossypium arboretum)中PPO1基因的抗蟲作用,本研究利用病毒介導的基因沉默(virus-induced gene silencing,VIGS),構(gòu)建基因沉默重組載體pTRV2-GhPPO1,轉(zhuǎn)化農(nóng)桿菌(Agrobacterium tumefaciens)GV3101,并侵染子葉展平的棉花植株;通過qRT-PCR檢測沉默效果,并用獲得的棉花植株飼喂二齡初期棉鈴蟲(Helicoverpa armigera)幼蟲,對沉默后棉花植株的抗蟲性及GhPPO1基因表達量的變化進行檢測。結(jié)果表明,利用煙草脆裂病毒(Tobacco rattle virus,TRV)體系成功抑制了棉花GhPPO1的表達,沉默后GhPPO1基因相對表達量僅為對照組(空載體pTRV0)表達量的20%(P0.05)。與飼喂正常棉花葉片的棉鈴蟲相比,飼喂GhPPO1基因沉默后棉花葉片的棉鈴蟲72 h后,體重增加86%,表明GhPPO1是棉花抗蟲物質(zhì)中的一類防御機制基因。棉鈴蟲直接誘導18、72 h的葉片中GhPPO1的表達量高于間接誘導葉片,證明沉默該基因后,消弱了棉花植株直接防御的能力,致使棉鈴蟲增重效果顯著。由此推測該基因可能參與棉花直接防御反應。本研究對于GhPPO1在棉花防御植食性昆蟲中的作用具有重要意義,為棉鈴蟲與棉花誘導防御之間的關(guān)系提供基礎(chǔ)資料。
[Abstract]:Polyphenol oxidase (PPO) is a kind of copper-containing redox enzyme. In order to study the insect-resistance of PPO1 gene in cotton Gossypium arboretum. In this study, virus-induced gene silencing (VIGSs) was used in virus-mediated gene silencing. The recombinant vector pTRV2-GhPPO1 was constructed and transformed into Agrobacterium tumefaciens)GV3101. And infecting cotton plants with flattened cotyledons; The silencing effect was detected by qRT-PCR, and the obtained cotton plants were fed with Helicoverpa armigera larvae. The insecticidal resistance and the expression of GhPPO1 gene in cotton plants after silencing were detected. The results showed that tobacco rattle virus was used for tobacco embrittlement virus. TRV) system successfully inhibited the expression of GhPPO1 in cotton. The relative expression of GhPPO1 gene after silencing was only 20% of that of the control group (empty vector pTRV0), compared with that of cotton bollworm fed normal cotton leaves. Cotton bollworm (Helicoverpa armigera) fed with GhPPO1 gene silencing for 72 h increased its weight by 86%. The results showed that GhPPO1 was a kind of defense gene in cotton insect-resistant materials, and the expression of GhPPO1 in leaves induced by Helicoverpa armigera for 1872 h was higher than that in indirectly induced leaves. It was proved that silencing the gene weakened the ability of cotton plants to defend directly. It is suggested that this gene may be involved in direct defense response of cotton. This study is of great significance for the role of GhPPO1 in cotton plant insect defense. To provide basic data for the relationship between cotton bollworm and cotton induced defense.
【作者單位】： 新疆農(nóng)業(yè)大學農(nóng)學院;中國農(nóng)業(yè)科學院棉花研究所/棉花生物學國家重點試驗室;
【基金】：國家轉(zhuǎn)基因生物新品種培育科技重大專項(No.2016ZX08012-004)
【分類號】：S435.62
【正文快照】： 棉花(Gossypium arboreum)是重要的經(jīng)濟作物,是紡織纖維、飼料等的重要來源。隨著棉花全基因組測序的完成,應用分子工具和技術(shù)資源分析基因功能至關(guān)重要(Wang et al.,2012)。棉花的生長周期長、基因組功能冗雜,遺傳轉(zhuǎn)化過程費力耗時并且通常很難實現(xiàn)(Gu et al.,2014)。病毒介，

本文編號：1458388