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中国农学通报 ›› 2015, Vol. 31 ›› Issue (32): 48-53.doi: 10.11924/j.issn.1000-6850.casb15050187

所属专题: 园艺

• 生物技术科学 • 上一篇    下一篇

黄瓜疫霉菌ITS及β-tubulin的扩增和序列分析

丁 婧1,吴晓金2,雷 颖1,易图永1,3,4   

  1. (1湖南农业大学植物保护学院,长沙 410128;2湖南省洪江市农业局,湖南黔城 418116;3植物病虫害生物学与防控湖南省重点实验室,长沙410128;4湖南省生物农药与制剂加工工程技术研究中心,长沙410128)
  • 收稿日期:2015-05-29 修回日期:2015-07-31 接受日期:2015-08-13 出版日期:2015-11-16 发布日期:2015-11-16
  • 通讯作者: 易图永
  • 基金资助:
    湖南省高校创新平台建设项目“黄瓜疫霉菌遗传多样性的AFLP分析”(13K064);农业部公益行业计划“蔬菜卵菌病害综合防控技术研究与示范”(201003004)。

Amplification and Sequence Analysis of ITS and β-tubulin from Phytophthora melonis

Ding Jing1, Wu Xiaojin2, Lei Ying1, Yi Tuyong1,3,4   

  1. (1College of Plant Protection, Hunan Agricultural University, Changsha 410128; 2Agricutural Bureau of Hongjiang City, Hunan Province, Qiancheng Hunan 418116; 3Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Plant Pests, Changsha 410128;4Hunan Provincial Engineering & Technology Research Center for Biopesticide and Formulation Processing, Changsha 410128)
  • Received:2015-05-29 Revised:2015-07-31 Accepted:2015-08-13 Online:2015-11-16 Published:2015-11-16

摘要: 为研究不同来源地黄瓜疫霉菌的系统发育关系以及遗传多样性,从湖南省内5个黄瓜种植区采集具有典型症状的黄瓜疫病病株分离纯化,利用形态学方法观察孢子囊形态。再运用PCR技术分别进行ITS以及β-tubulin扩增测序并与Genbank的中相关序列构建系统发育树。经形态学初步鉴定22株黄瓜疫霉菌株,ITS-PCR序列长度约为780 bp,β-tubulin-PCR序列长度约为800 bp,所有测得序列与下载自Genbank的相关序列存在不同程度的相似性。ITS序列的系统发育分析,用于构建系统发育树的序列分为2个大的聚类I和II,黄瓜疫霉菌均位于I组内,由于来源地差异,不同地区的黄瓜疫霉菌又被聚在不同小类;β-tubulin序列在系统发育分析中被分为4个聚类,22个P. melonis序列与下载的P. melonis序列均在聚类组I中。聚类分析结果表明:疫霉属种群间具有明显的亲缘关系,不同地域的黄瓜疫霉菌遗传多样性表现出差异,但总体趋势是相同来源地的菌株亲缘性比较接近。

关键词: 环县, 环县, 气温, 降水, 干旱

Abstract: For researching the genetic diversity and phylogenetic relationships of Phytophthora melonis, infected plants with classical symptoms of cucumber phytophthora blight were collected from 5 different cucumber growing areas in Hunan Province. P. Melonis isolated and purified from diseased cucumbers were preliminarily identified by morphological method of observing the sporangia, then ITS and β-tubulin of 22 strains were amplified using PCR technology and sequenced, these sequenced strains were aligned with related sequences downloaded from Genbank with bioinformatics software. The sequence length of ITS-PCR was about 780 bp, and β-tubulin-PCR was about 800 bp. There was similarity between the sequenced strains and the related sequences downloaded from Genbank. According to the phylogenetic analysis of ITS sequence, the sequences used to construct phylogenetic tree were divided into two large clusters: group I and group II, the sequenced P. melonis was located in group I. Since the sources were different, 22 strains from different areas were gathered in different small branches. According to phylogenetic analysis of β-tubulin sequences, the sequences were divided into four big clusters, 22 P. melonis sequences and downloaded P. melonis sequences were in the group I. The results of classification indicated that the two methods were able to reflect the affinity of different phytophthora, and strains isolated from different areas had genetic diversity, but the same source isolates had close genetic relationship.