TRV病毒介導(dǎo)的基因沉默體系在新疆陸地棉和亞洲棉中的建立
發(fā)布時間:2018-04-21 17:03
本文選題:Gossypium + arboreum ; 參考:《棉花學(xué)報》2016年05期
【摘要】:為研究煙草脆裂病毒(Tobacco rattle virus,TRV)介導(dǎo)的基因沉默(Virus-induced gene silencing,VIGS)技術(shù)應(yīng)用的廣譜性,以陸地棉(Gossypium hirsutum)CLA1為標(biāo)記基因,通過構(gòu)建基因沉默載體,在新陸早33和亞洲棉(Gossypium arboreum)建立了TRV-VIGS體系。半定量PCR分析表明,與對照相比,TRV侵染的新陸早33和亞洲棉中CLA1基因的表達(dá)均被有效抑制,真葉表現(xiàn)出明顯的白化癥狀。同時在45份新陸中系列棉花材料中重復(fù)該體系,并且通過控制培養(yǎng)溫度來檢驗溫度對VIGS的影響,結(jié)果表明,當(dāng)晝/夜溫度分別為23℃/21℃和32℃/28℃時,不同品種的新陸中系列棉花均可以維持不同程度的白化表型。證明,TRV介導(dǎo)的VIGS可以用于室內(nèi)研究新陸中系列棉花和亞洲棉基因功能。
[Abstract]:In order to study the broad-spectrum application of Virus-induced gene silencing (VIGS) mediated by Tobacco rattle virus (TMV), the TRV-VIGS system was established in Xinluzao 33 and Gossypium arboreum by constructing gene silencing vector using Gossypium hirsutum)CLA1 as marker gene. Semi-quantitative PCR analysis showed that the expression of CLA1 gene was effectively inhibited in Xinluzao 33 and Asian cotton, and the true leaves showed obvious albinism symptoms. At the same time, the system was repeated in 45 cotton materials, and the effect of temperature on VIGS was tested by controlling the culture temperature. The results showed that the temperature of day / night was 23 鈩,
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