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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (25): 150-153.

所属专题： 园艺

• 林学 园艺 园林 • 上一篇    下一篇

黄瓜细菌性角斑病的分子检测

王平 樊金娟 刘长远 赵奎华 梁春浩   

  • 收稿日期:2011-10-08 修回日期:2011-12-21 出版日期:2012-09-05 发布日期:2012-09-05
  • 基金资助:

    辽宁省百千万人才项目

The Detection of Pseudomonas syringae pv. Based on PCR

  • Received:2011-10-08 Revised:2011-12-21 Online:2012-09-05 Published:2012-09-05

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摘要：

探索黄瓜细菌性角斑病菌(Pseudomonas syringae pv.)的分子检测技术，为快速、准确鉴定和检测提供技术和方法。根据丁香假单胞杆菌黄瓜角斑病致病型病菌与其他黄瓜病原菌核糖体基因转录间隔区 (16S-23S Ribosomal DNA Intergenic Spacer，ITS)序列间的差异，设计特异性引物PLf1/PLr2 (PLf1: 5'-ATA AGG GTG AGG TCG GCA GTT-3'，PLr2: 5'-CTC GTC TTT CAT CGC CTT TG-3')。引物PLf1/PLr2可从丁香假单胞杆菌黄瓜角斑病致病型病菌中增出一条473 bp的条带，将黄瓜细菌性角斑病菌和其他菌种分开。建立的黄瓜细菌性角斑病分子检测的方法可用于该病害的快速分子检测，可直接对黄瓜叶片汁液进行病原菌检测，并且可在接种后病症发生前72 h内检测到病原菌。

关键词: 区域, 区域

Abstract:

The aim was to study the detection technology of Pseudomonas syringae pv. in cucumber, and provide the method for quick and precise detection. According to the differences in 16S-23S Ribosomal DNA Intergenic Spacer（ITS）sequences of Pseudomonas syringae pv. and other cucumber pathogens, a couple of specific primers of PLf1/PLr2（PLf1: 5' ATA AGG GTG AGG TCG GCA GTT 3', PLr2: 5' CTC GTC TTT CAT CGC CTT TG 3'）were synthesized. Using PLf1/PLr2 primers, only a single PCR band of 473 bp from Pseudomonas syringae pv. was amplified and no PCR band from other species. The primers could identify and distinguish Pseudomonas syringae pv. from other cucumber pathogens. The PCR-based method could be used for the accurate and rapid identification of Pseudomonas syringae pv. Furthermore, the pathogenic bacteria were detected by this diagnostic technique in 72h before syptome was observational.