發(fā)布時(shí)間：2021-07-09 13:09

　　該研究以抗病品種中國(guó)野生毛葡萄‘商-24’和感病品種歐洲葡萄‘紅地球’為材料,利用RT-PCR方法克隆TLP15基因,分別命名為VqTLP15和VvTLP15（GSVIVT01018769001）,對(duì)其進(jìn)行生物信息學(xué)分析、亞細(xì)胞定位、轉(zhuǎn)化擬南芥,并接種不同病原菌觀察分析轉(zhuǎn)基因株系的抗性,采用qRT-PCR檢測(cè)SA和JA/Eth信號(hào)途徑以及調(diào)控氣孔運(yùn)動(dòng)的相關(guān)基因表達(dá)。結(jié)果顯示:（1）成功克隆獲得VqTLP15基因的開放閱讀框（ORF）;氨基酸序列比對(duì)顯示,VqTLP15基因與葡萄基因組網(wǎng)站歐洲葡萄‘黑比諾’VvTLP15和‘紅地球’克隆的VvTLP15基因的同源性分別為98.99%和99.66%。（2）亞細(xì)胞定位表明,VqTLP15定位于細(xì)胞質(zhì)。（3）成功獲得VqTLP15轉(zhuǎn)基因擬南芥株系（L1、 L2、 L3）。（4）接種觀察發(fā)現(xiàn):白粉菌處理7 d后轉(zhuǎn)基因株系對(duì)白粉菌的抗性較野生型（Col-0）提高,且其葉片的白粉菌孢子濃度顯著低于Col-0;灰霉菌誘導(dǎo)的葉片壞死性損傷在轉(zhuǎn)基因株系（L1、L2和L3病斑面積>40%的比例分別為71%、62%和67%）中顯著大于Col-0（43%）... 

【文章來(lái)源】：西北植物學(xué)報(bào). 2020,40(05)北大核心CSCD

【文章頁(yè)數(shù)】：12 頁(yè)

【文章目錄】：
1 Materials and methods
    1.1 Plant materials
    1.2 Methods
        1.2.1 VqTLP15 sequence analysis
        1.2.2 Subcellular localization analysis of VqTLP15
        1.2.3 Generation of VqTLP15 over-expressing A. thaliana lines
        1.2.4 A. thaliana disease assays
        1.2.5 Histochemical staining
        1.2.6 Response of stomata to different treatments
        1.2.7 RNA isolation and quantitative RT-PCR
    1.3 Statistical analysis
2 Results
    2.1 VqTLP15 cloning and sequence analysis
    2.2 Subcellular localization of VqTLP15
    2.3 Response of VqTLP15 over-expressing A. thaliana lines to powdery mildew
    2.4 Response of VqTLP15 over-expressing A. thaliana lines to B. cinerea
    2.5 Response of VqTLP15 over-expressing A. thaliana lines to PstDC3000
    2.6 Stomatal closure immunity response
3 Discussion



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