CAJ | 학술논문

试验以瓜叶菊（Pericallis hybrida B. Nord.）为试验材料，采用RT-PCR和RACE方法获得白粉菌调控相关Mlo基因的cDNA全长序列，通过VIGS技术构建载体，进行基因沉默并转入到农杆菌GV3101中。序列分析表明，该基因cDNA全长包含1个1296 bp的开放阅读框，编码431个氨基酸。荧光定量结果表明，Mlo基因在瓜叶菊的根、茎、叶中都有表达，叶片中表达量最高，根中表达量最低。根据PCR结果和双酶切鉴定结果表明载体构建成功。通过PCR结果显示载体已成功转入农杆菌GV3101中。该研究为下一步分析基因沉默后瓜叶菊中Mlo基因变化提供依据。
시험이과협국（Pericallis hybrida B. Nord.）위시험재료，채용RT-PCR화RACE방법획득백분균조공상관Mlo기인적cDNA전장서렬，통과VIGS기술구건재체，진행기인침묵병전입도농간균GV3101중。서렬분석표명，해기인cDNA전장포함1개1296 bp적개방열독광，편마431개안기산。형광정량결과표명，Mlo기인재과협국적근、경、협중도유표체，협편중표체량최고，근중표체량최저。근거PCR결과화쌍매절감정결과표명재체구건성공。통과PCR결과현시재체이성공전입농간균GV3101중。해연구위하일보분석기인침묵후과협국중Mlo기인변화제공의거。
In this paper, Mlo gene full length cDNA was obtained by reverse transcription polymerase chain reaction (RT-PCR) and RACE, then through VIGS vector gene silencing technology to build and transferred into Agrobacterium GV3101. The result of sequence analysis indicated that the Mlo gene from Pericallis hybrida B. Nord contained about 1 296 bp, encoding 431 amino acids. Finally, through fluorescence quantitative analysis, we found that Mlo gene showed the highest expression level in leaves and the lowest in root. According to PCR results and double digestion results showed the vector was successfully constructed. PCR results showed that the carrier through the broth had been successfully transferred to Agrobacterium GV3101. The research for the next step in the analysis of gene silencing cineraria provides the basis for changes in Mlo gene.