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中国农学通报 ›› 2012, Vol. 28 ›› Issue (9): 163-168.

所属专题: 烟草种植与生产

• 生物技术科学 • 上一篇    下一篇

一种土壤中烟草疫霉菌的快速分离鉴定及计数方法

王晗   

  • 收稿日期:2011-09-09 修回日期:2011-12-23 出版日期:2012-03-25 发布日期:2012-03-25
  • 基金资助:

    四川省烟草专卖局项目

A Fast Method of Isolation, Identification and Counting of Phytophthora nicotianae from Soil

  • Received:2011-09-09 Revised:2011-12-23 Online:2012-03-25 Published:2012-03-25

摘要:

烟草疫霉菌是一种常见的土传植物病原菌,其所导致的烟草黑胫病对烟草业造成了巨大经济损失。准确统计土壤中烟草疫霉菌的数量,有助于了解土壤中烟草疫霉菌的分布,密度以及发生动态,进而可以及时发布灾情预报,减少经济损失。本研究采用土壤稀释平板法,在选择性培养基上较好的对烟草疫霉菌进行了分离;运用微量真菌DNA提取方法成功提取了平板上菌株的DNA;采用特异引物,对提取的7个疑似菌株DNA进行PCR验证,结果有6个菌株扩增出了737bp的烟草疫霉菌特异性目的基因片段;ITS进一步验证6个菌株与已报道烟草疫霉菌相似度达99%以上;表明6个疑似菌株是烟草疫霉菌。该方法具有相对准确、低成本、简单易推广的优点。

关键词: 重金属, 重金属

Abstract:

Phytophthora nicotianae is a conventional soil-borne plant pathogens,which induces tobacco black shank and causes great financial loss to tobacco industry. Accurate quantity analysis of P. nicotianae in soil helps researchers acquire their distribution, density and dynamics,provide information for disaster forecast and minimizes the economic losses. Pour plate method was used to separate the fungi on the selective medium,and then DNA of the fungal strain was extracted by the method for mini-preparation of fungal DNA from the plate. After that the selected 7 strains were tested by PCR with specific primers. The results showed that specific gene fragments of P. nicotianae with the length of 737bp were obtained from 6 of 7 strains. Further ITS testing found their gene sequence to be in good agreement with that of the reported Phytophthora nicotianae by 99%. Those proved the 6 strains were P. nicotianae in certain. This method has advantages over others in accuracy, cost and ease for application.

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