發(fā)布時(shí)間：2018-02-20 21:25

  本文關(guān)鍵詞： 病毒誘導(dǎo)的基因沉默 黃瓜花葉病毒 b 八氫番茄紅素脫氫酶　出處：《浙江農(nóng)業(yè)學(xué)報(bào)》2017年04期 　論文類(lèi)型：期刊論文

【摘要】：病毒誘導(dǎo)的基因沉默(virus-induced gene silencing,VIGS)已被廣泛應(yīng)用于植物基因的功能研究。試驗(yàn)以八氫番茄紅素脫氫酶(phytoene desaturase,PDS)作為指示基因,在本生煙體內(nèi)分析黃瓜花葉病毒(Cucumber mosaic virus,CMV)作為VIGS載體對(duì)植物內(nèi)源PDS mRNA的沉默效率。采用分子克隆的方法,將FnyCMV 2b ORF替換為不同長(zhǎng)度的PDS序列,并分析了其在本生煙植物上誘導(dǎo)的PDS沉默效率。結(jié)果表明,該重組病毒能誘導(dǎo)上部系統(tǒng)葉的光漂白表型,且沉默效率與插入片段長(zhǎng)度呈一定的正相關(guān)性,但在頂端新生葉中均不引起明顯的光漂白現(xiàn)象,過(guò)長(zhǎng)的片段插入(≥518 nt)影響病毒基因組的遺傳穩(wěn)定性,導(dǎo)致插入片段的部分缺失。CMV作為VIGS載體,最適合的插入片段約為350 nt。
[Abstract]:Virus-induced gene silencing (VIGS) has been widely used to study the function of plant genes. Phytoene desaturase (PDS) was used as an indicator gene. The silencing efficiency of cucumber mosaic virus (Cucumber mosaic virusus) to plant endogenous PDS mRNA was analyzed in vivo. FnyCMV 2b ORF was replaced with different length PDS sequences by molecular cloning. The results showed that the recombinant virus could induce the photobleaching phenotype of the upper leaves, and the silencing efficiency was positively correlated with the length of the inserted fragment. However, there was no obvious photobleaching in the apical neonate leaves. The long fragment insertion (鈮，

本文編號(hào)：1520062