https://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
https://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
https://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
https://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2010, Vol. 37 ›› Issue (10): 1567-1574.

• 果树 • 上一篇    下一篇

抗病基因VpPR10转化‘砀山酥梨’及转化条件的优化

孟颢光1,2,3,4,张朝红1,2,3,王跃进1,2,3,*   

  1. (1西北农林科技大学园艺学院,陕西杨凌 712100;2农业部西北园艺植物种质资源利用重点开放实验室,陕西杨凌 712100;3陕西省农业分子生物学重点实验室,陕西杨凌 712100;4河南农业大学植物保护学院,郑州 450002)
  • 收稿日期:2010-05-26 修回日期:2010-08-03 出版日期:2010-10-25 发布日期:2010-10-25
  • 通讯作者: 王跃进

Transforming Diseases-resistant Gene VpPR10 into Pear‘Dangshan Suli’and Optimization of the Transformation System

MENG Hao-guang1,2,3,4,ZHANG Chao-hong1,2,3,and WANG Yue-jin1,2,3,*   

  1. (1College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100,China;2Key Laboratory of Horticultural Plant Germplasm Resources Utilization in Northwest China,Ministry of Agriculture of China,Yangling,Shaanxi 712100,China;3Shaanxi Key Laboratory for Molecular Biology of Agriculture,Yangling,Shaanxi 712100,China;4College of Plant Protection,Henan Agricultural University,Zhengzhou 450002,China)
  • Received:2010-05-26 Revised:2010-08-03 Online:2010-10-25 Published:2010-10-25
  • Contact: WANG Yue-jin

摘要: 将源自中国野生华东葡萄的抗病基因——病程相关蛋白基因导入‘砀山酥梨’,并对转化系统条件进行优化。首先将VpPR10基因重组于中间表达载体pWR-Ⅱ,得到重组质粒pWR-Ⅱ-VpPR10;继而以改进冻融法将其导入根癌农杆菌EHA105,再利用根癌农杆菌介导的叶盘转化法进行遗传转化。根癌农杆菌浓度OD600 = 0.5,离体叶片侵染5 min,在含有100 μmol · L-1乙酰丁香酮的培养基中暗处共培养48 h有利于砀山酥梨的遗传转化;经PCR分析、Southern blot和RT-PCR检测,证明目的基因VpPR10已导入并整合到砀山酥梨基因组中;转化率为1.08%。

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

中图分类号: