Overexpression of VpPR10.1 by an efficient transformation method enhances downy mildew resistance in V-vinifera
Overexpression of VpPR10.1 by an efficient transformation method enhances downy mildew resistance in V-vinifera
复制标题
通过有效转化方法过度表达 VpPR10.1 可增强葡萄树的霜霉病抗性
DOI：
10.1007/s00299-018-2271-z
复制
发表时间：
2018-05-01
影响因子：
6.2
通讯作者：
Xu, Yan
中科院分区：
文献类型：
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作者：
Su, Hang;Jiao, Yun-Tong;Xu, Yan
腐胺和亚精胺提高了葡萄属葡萄的转化效率。CV.汤普森无籽。转基因无核葡萄中VpPR10.1的积累可能会增加其对霜霉病的抗性。描述了一种更有效的促进农杆菌介导的葡萄转化的方法。CV.利用多胺（PAs）进行Thompson无核体细胞胚胎发生。腐胺、亚精胺和精胺的效力在体细胞胚生长期间加入培养基的浓度范围（10 μ M、100 μ M和1mM）下鉴定。腐胺（PUT）和亚精胺（SPD）促进共培养后原胚团（PEM）的恢复和体细胞胚（SE）的发育。从时间框架在遗传转化中的重要性来看，在共培养阶段添加多胺比延迟选择处理具有更强的效果，在选择阶段上级抗生素处理。最佳的胚发生反应是在共培养基中加入1mMPUT和100 μ M SPD。利用上述方法，将中国野生葡萄属葡萄病程相关基因VpPR 10. 1导入无核白葡萄中，并对其进行功能分析。将通过蛋白质印迹分析确认的转基因株系接种葡萄霜霉病菌以测试霜霉病抗性。基于观察到的菌丝生长的限制和H2O2积累的增加，VpPR 10.1的积累可能增强了转基因植物对霜霉病的抗性。
Putrescine and spermidine increase the transformation efficiency of Vitis vinifera L. cv. Thompson seedless. Accumulation of VpPR10.1 in transgenic V. vinifera Thompson seedless, likely increases its resistance to downy mildew.A more efficient method is described for facilitating Agrobacterium-mediated transformation of Vitis vinifera L. cv. Thompson Seedless somatic embryogenesis using polyamines (PAs). The efficacies of putrescine, spermidine and spermine are identified at a range of concentrations (10 A mu M, 100 A mu M and 1 mM) added to the culture medium during somatic embryo growth. Putrescine (PUT) and spermidine (SPD) promote the recovery of proembryonic masses (PEM) and the development of somatic embryos (SE) after co-cultivation. Judging from the importance of the time-frame in genetic transformation, PAs added at the co-cultivation stage have a stronger effect than delayed selection treatments, which are superior to antibiotic treatments in the selection stage. Best embryogenic responses are with 1 mM PUT and 100 A mu M SPD added to the co-culture medium. Using the above method, a pathogenesis-related gene (VpPR10.1) from Chinese wild Vitis pseudoreticulata was transferred into Thompson Seedless for functional evaluation. The transgenic line, confirmed by western blot analysis, was inoculated with Plasmopara viticola to test for downy mildew resistance. Based on observed restrictions of hyphal growth and increases in H2O2 accumulation in the transgenic plants, the accumulation of VpPR10.1 likely enhanced the transgenic plants resistance to downy mildew.