RNAi和 GroEL 介导的双价转基因番茄对 TYLCV的抗性

摘要

Tomato yellow leaf curl virus ( TYLCV) is one of the most devastating viruses of cultivated tomatoes in the world. Recently, the disease occurred suddenly in China, spreading towards north from south, which has resulted in great economic losses. The lack of resistance resource limits the TYLCV resistance breeding. In order to generate engineered re-sistance, the plant expression vectors SUC2-GroEL and pBI121-AC1-AC2 were co-transformed into tomato by agrobacterium-mediated genetic transformation method, and nineteen kanamycin resistant regenerated tomato plants were obtained, inclu-ding five plants of SUC2-GroEL vector, three plants of pBI121-AC1-AC2 vector and three plants of double vectors confirmed by PCR and RT-PCR. T0 generation of the transgenic plants were inoculated with TYLCV, and it turned out 8 were TYLCV resistant. The carrying rates of TYLCV by T1 generation achieved via vectors SUC2-GroEL, pBI121-AC1-AC2, and SUC2-GroEL/pBI121-AC1-AC2 were 35%, 25%, and 15%, respectively, suggesting that the plants with double genes have a stronger disease resistance than other plants with single gene.%本研究将已构建的SUC2-GroEL和pBI121-AC1-AC2植物表达载体共转化番茄，共获得19株卡那霉素抗性再生植株。经过PCR和RT-PCR检测，初步获得转SUC2-GroEL载体的阳性植株5株，转pBI121-AC1-AC2载体的阳性植株3株以及转SUC2-GroEL/ pBI121-AC1-AC2双载体的阳性植株2株。利用TYLCV侵染性克隆农杆菌注射接种法对T0代转基因阳性植株进行抗性鉴定，获得抗TYLCV的转基因植株7株。对T0代未扩出病毒条带的T1代植株进行抗性鉴定，结果发现转SUC2-GroEL、pBI121-AC1-AC2、SUC2-GroEL/pBI121-AC1-AC2不同载体的株系植株中平均带毒率依次是35%、25%和15%，推断转双基因的植株的抗病能力优于转单个基因植株的抗病能力。