西红柿原系统素基因转化水稻的抗虫性研究

构建了两个含西红柿原系统素基因的双元载体pNAR304（UbiI5’+Prosystemin+NOS3’）和pNAR305 (UbiI5’+Prosystemin+NOS3’+ PinⅡ5’+PinⅡ+PinⅡ3’)，并用农杆菌介导方法将其转入水稻品种秀水63、合江19和日本晴。经潮霉素抗性、PCR和Southern blot确证，共获得转基因水稻14株。Northern blot检测表明，原系统素基因(Prosystemin)和马铃薯蛋白酶抑制剂基因(PinⅡ)在这些转基因植株中都能转录表达。然而，植株的二化螟和褐飞虱抗虫性鉴定表明：单独转入Prosystemin（pNAR304）不能提高转基因水稻的抗虫能力；Prosystemin和PinⅡ双价（pNAR305）转基因植株能明显提高水稻的抗虫性，但其抗性水平与PinⅡ单个基因的转基因水稻植株间并无显著差异。 这表明，Prosystemin基因转入水稻并不能有效调节转基因水稻PinII基因的表达量。据此试验结果推测，水稻中可能不存在类似于西红柿系统素的信号途径，很可能水稻的伤害信号转导是经由与双子叶植物系统素体系不同的其它途径来实现的。

Insect Resistance Study for Transgenic Rice with Tomato prosystemin Gene

Two binary vectors containing tomato presystemin gene pNAR304（UbiI5’+Prosystemin+NOS3’）and pNAR305 (UbiI5’+Prosystemin+NOS3’ + PinⅡ5’+PinⅡ+PinⅡ3’) were constructed and were introduced into rice cvs. Xiushui63, Hejiang19 and Nipponbare by Agrobacterium-mediated transformation. From tests of hygromycin resistance, PCR and Southern blot, 14 independent transgenic plants were confirmed. Northern blot analysis indicated that both presystemin gene and PinⅡ gene could transcribed in these transgenic plants. But in the rice insect resistance bioassays to stripe stem borer (Chilo suppressalis) and brown plant hopper (Nilaparvata luguns stal), insect resistance of those plants transformed by prosystemin gene alone（pNAR304） showed no improvement than control plants and insect resistance of transgenic plants carrying both prosystemin gene and PinⅡgene（pNAR305）exhibited significant increase than control plants, but their insect resistance level was similar to those transgenic plants only could not regulate the expression of PinⅡgene in transgenic rice. From above experiment results, we think that there may be no signal pathway for tomato systemin in the rice and, probably, signal transmission of injury in rice were other pathway different from dicotyledonous systemin.