| 应用RT-PCR和斑点杂交法检测新疆梨树上的苹果锈果类病毒 |
| |
| 引用本文： | 赵英,牛建新.应用RT-PCR和斑点杂交法检测新疆梨树上的苹果锈果类病毒[J].果树学报,2007,24(6):761-764. |
| |
| 作者姓名： | 赵英 牛建新 |
| |
| 作者单位： | 石河子大学农学院园艺系,新疆石河子,832003 |
| |
| 基金项目： | 国家自然科学基金
,
国家科技攻关项目
,
兵团科委资助项目 |
| |
| 摘 要： | 利用自行设计的方法分离提取总RNA,通过一对特异引物,分别扩增出不同梨树品种上苹果锈果类病毒的特异片段,通过对库尔勒香梨品种上获得的特异片段回收、克隆和测序。结果表明,该片段长250bp,与已发表的AF421195同源性为99%,在此基础上利用克隆的ASSVd质粒,通过RT-PCR合成了生物素标记的cDNA探针,利用该探针对梨样品进行了斑点杂交检测,取得了很好的检测效果。进一步证明该反应体系能很好地用于梨树苹果锈果类病毒的RT-PCR检测。
|
| 关 键 词： | 梨树 新疆 苹果锈果类病毒 RT-PCR cDNA探针 测序 |
| 文章编号： | 1009-9980（2007）06-761-04 |
| 修稿时间： | 2007-06-08 |
Detection of apple scar skin viroid in pear cultivars by RT-PCR and spot hybridization in Xinjiang area |
| |
| ZHAO Ying,NIU Jian-xin.Detection of apple scar skin viroid in pear cultivars by RT-PCR and spot hybridization in Xinjiang area[J].Journal of Fruit Science,2007,24(6):761-764. |
| |
| Authors: | ZHAO Ying NIU Jian-xin |
| |
| Institution: | Department of Horticulture, College of Agriculture, Shihezi University, Shihezi, Xinjiang 832003 China |
| |
| Abstract: | Total RNA was isolated using the method designed by ourselves. Full-length fragments of Apple scar skin viroid (ASSVd) have been amplified by one specific pair of primer from different pear samples. The ASSVd -specific fragment isolated from Korla’s Xiangli pear cultivar has been recovered,cloned and sequenced,and the result showed its length was 250 bp representing the homology of 99% compared with AF421195 published on NCBI database. Then biotin-labelled cDNA probe was synthesized using cloned plasmids by RT-PCR. The probe has been used well for spot hybridization detection of ASSVd,which further proved RT-PCR can be successfully used for the detection of ASSVd in pear. |
| |
| Keywords: | Pear Xinjiang area ASSVd RT-PCR cDNA probe Sequence |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
