关键词: 梨, 抗病基因, 农杆菌介导, 病程相关蛋白基因（VpPR10）, 条件优化

Abstract: To provide the basis for increasing the genetic transformation efficiency of Pyrus bretshneideri‘Dangshan Suli’，the VpPR10 gene，cloned from Chinese wild Vitis pseudoreticulata W. T. Wang，was introduced into pear genome and the optimum conditions of the genetic transformation system were also examined. The VpPR10 gene was constructed into the plant expression vector pWR-Ⅱ and the recombinant plasmid pWR-Ⅱ-VpPR10 was obtained；Then the pWR-Ⅱ-VpPR10 was introduced into the Agrobacterium tumefaciens strain EHA105 by improved freeze-thaw method，and was transformed into leaf disc explants of Dangshan Suli via an Agrobacterium tumefaciens-mediated system. The results showed that the suitable transformation conditions were 5 min infection time，0.5 bacterium concentration （OD600），100 μmol · L-1 Acetosyringone（AS），and 48 h co-cultivation in dark. The PCR，Southern blot and RT-PCR analysis results revealed that VpPR10 gene was successfully recombined into the genome of Dangshan Suli，and the rate of genetic transformation was 1.08%.

Key words: Pyrus bretschneideri Rehd., disease-resistant gene, Agrobacterium tumefaciens- mediated, pathogenesis-related protein gene（VpPR10）, optimization