拟南芥抗根结线虫和蚜虫的功能基因鉴定

摘要

To identify plant genes that can increase resistance towards root knot nematode ( RKN) and aphid, nine Arabidopsis activation tag mutants carrying aphid resistance were used to explore RKN resistance. Inverse PCR was per⁃formed to locate the activation tag on Arabidopsis genome and quantitative RT⁃PCR was used to quantify the expression lev⁃els of upstream and downstream genes. Three out of the nine aphid resistant mutants were identified with resistance towards RKN, indicated by lower infection level compared to wild type at 7 days after inoculation. Among these three mutants, two had almost the same location of the activation tag and activated SKS13 gene. In the other mutant, activation tag enhanced the expression level of PERK2 gene. Transgenic lines overexpressing either SKS13 or PERK2 resulted in reduced infection level of RKN and poorer aphid performance.%为了鉴定出增强植物对根结线虫和蚜虫抗性的功能基因，从9个抗蚜虫的拟南芥激活标签突变体中筛选增强根结线虫抗性的突变体。利用反向PCR技术定位激活标签在基因组上的插入位点，并通过荧光定量PCR技术分析插入位点上、下游基因表达水平，结合根结线虫和蚜虫在过量表达转基因植株上的表现验证基因功能。结果显示接种根结线虫后7 d，侵入3个拟南芥突变体根系的根结线虫数量明显少于野生型。其中2个突变体中的激活标签插入位点只相差2个碱基，SKS13基因表达水平提高；另一个株系中激活标签导致PERK2基因的表达水平提高。与野生型植株相比，分别过量表达SKS13和PERK2的转基因植株能明显减少侵入根系的根结线虫数量，同时蚜虫的繁殖数量也较低。