[与仙人掌丛枝病相关的植原体CWB1菌株16S rRNA基因的克隆与测序]

[Cloning and sequencing of 16S rRNA gene of Phytoplasma CWB1 strain associated with cactus witches' broom].

作者信息

Cai H, Li F, Kong B, Chen H

机构信息

Key Laboratory of Plant Pathology of Yunnan Province, Yunnan Agricultural University, Kunming 650201.

出版信息

Wei Sheng Wu Xue Bao. 2001 Dec;41(6):693-8.

PMID:12552825

Abstract

A 1.5 kb DNA fragment was amplified in DNA samples extracted from Opuntia salmiana porm showed witches'-broom symptom. The result indicates the existence of phytoplasma associated with this disease and this phytoplasma was designated as CWB1. The amplified fragment was ligated to pGEM-T easy vector and then transformed into JM109 strain of E. coli. Cloned DNA fragments were verified by PCR, restriction endonuclease (EcoRI) digestion and sequence analysis. The result revealed that the 16S rRNA gene of CWB1 consists of 1489 bp and shared 99.7% homology with Faba bean phyllody which belongs to phytoplasma 16S rII-C subgroup. So we can classify this strain into phytoplasma 16S rII-C subgroup.

摘要

从表现出丛枝病症状的肉质仙人掌（Opuntia salmiana porm）中提取的DNA样本中扩增出一个1.5 kb的DNA片段。结果表明存在与该病相关的植原体，该植原体被命名为CWB1。将扩增片段连接到pGEM-T easy载体上，然后转化到大肠杆菌JM109菌株中。通过PCR、限制性内切酶（EcoRI）消化和序列分析对克隆的DNA片段进行验证。结果显示，CWB1的16S rRNA基因由1489 bp组成，与属于植原体16S rII-C亚组的蚕豆叶变病植原体具有99.7%的同源性。因此，我们可以将该菌株归类为植原体16S rII-C亚组。