Kollár A, Thole V, Dalmay T, Salamon P, Balázs E
Agricultural Biotechnology Center, Institute for Plant Sciences, Gödöllö, Hungary.
Biochimie. 1993;75(7):623-9. doi: 10.1016/0300-9084(93)90069-5.
The coat protein (CP) gene from potato virus Y (Hungarian isolate, PVY-H) was engineered into Agrobacterium tumefaciens binary vector for expression in different tobacco lines. Three different Nicotiana tabacum breeding lines were transformed and the integration of the CP gene was confirmed by PCR technique using genomic DNA preparations. The transcription and expression of the integrated CP gene was detected by Northern and Western blots. Pathogen-derived resistance was demonstrated by inoculation of the R1 progeny of the transformed lines with purified PVY-H. The efficiency of protection varied between different transgenic plants ranging from almost complete to no protection. Five CP expressing tobacco lines were resistant to challenge infection with PVY-H as indicated by attenuation or absence of symptom development associated with reduction or lack of detectable virus accumulation. Data from Western blots showed that there is no correlation between the level of the expressed CP and the extent of protection. This suggests that the mechanism of the observed resistance is independent of the level of CP accumulation in the transgenic tobacco plants.
将马铃薯Y病毒（匈牙利分离株，PVY-H）的外壳蛋白（CP）基因构建到根癌农杆菌双元载体中，以便在不同烟草品系中表达。对三个不同的烟草育种品系进行了转化，并使用基因组DNA制备物通过PCR技术确认了CP基因的整合。通过Northern杂交和Western印迹检测整合的CP基因的转录和表达。用纯化的PVY-H接种转化品系的R1后代，证明了病原物衍生抗性。不同转基因植物之间的保护效率各不相同，从几乎完全保护到没有保护。五个表达CP的烟草品系对PVY-H的挑战感染具有抗性，表现为症状发展减弱或没有症状发展，同时病毒积累减少或无法检测到。Western印迹数据表明，表达的CP水平与保护程度之间没有相关性。这表明观察到的抗性机制与转基因烟草植物中CP的积累水平无关。
