土壤中烟草根黑腐病菌的实时定量PCR检测技术研究

康振辉, 黄俊丽

植物病理学报 ›› 2010, Vol. 40 ›› Issue (2) : 210-213.

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PDF(264 KB)
植物病理学报 ›› 2010, Vol. 40 ›› Issue (2) : 210-213.
研究简报

土壤中烟草根黑腐病菌的实时定量PCR检测技术研究

  • 康振辉, 黄俊丽
作者信息 +

Detection of Thielaviopsis basicola in soil with real-time quantitative PCR

  • KANG Zhen-hui, HUANG Jun-li
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文章历史 +

摘要

Thielaviopsis basicola is a soil-borne plant pathogen which causes root rot disease in tobacco plants. Detection and monitoring of T. basicolain soil is of great significance to control this disease. Based on the differences in internal transcribed spacer (ITS) sequences of T. basicola and other fungal pathogens, a specific primer pair Tb1/Tb2 for T. basicolawas developed. The results showed that the primer pair gave a single amplicon of 330 bp from T. basicola and revealed no undesirable cross-reaction with other seven soil-borne pathogen isolates and three tobacco rhizosphere dominant fungi isolates. With a series of 10-fold genomic DNA dilutions of T. basicola, the detection limit of 1 pg/μL in conventional PCRand100 fg/μL in real-time quantitative PCR was achieved. With DNA from the soil inoculated with different numbers of T. basicola conidia, the detection limit was 10 conidia per reaction in conventional PCR and 0.4 conidia per reaction in real-time quantitative PCR.

Abstract

Thielaviopsis basicola is a soil-borne plant pathogen which causes root rot disease in tobacco plants. Detection and monitoring of T. basicolain soil is of great significance to control this disease. Based on the differences in internal transcribed spacer (ITS) sequences of T. basicola and other fungal pathogens, a specific primer pair Tb1/Tb2 for T. basicola was developed. The results showed that the primer pair gave a single amplicon of 330 bp from T. basicola and revealed no undesirable cross-reaction with other seven soil-borne pathogen isolates and three tobacco rhizosphere dominant fungi isolates. With a series of 10-fold genomic DNA dilutions of T. basicola, the detection limit of 1 pg/μL in conventional PCRand100 fg/μL in real-time quantitative PCR was achieved. With DNA from the soil inoculated with different numbers of T. basicola conidia, the detection limit was 10 conidia per reaction in conventional PCR and 0.4 conidia per reaction in real-time quantitative PCR.

关键词

Thielaviopsis basicola / internal transcribed spacer (ITS) / real-time quantitative PCR / soil

Key words

Thielaviopsis basicola / internal transcribed spacer (ITS) / real-time quantitative PCR / soil

引用本文

导出引用
康振辉, 黄俊丽. 土壤中烟草根黑腐病菌的实时定量PCR检测技术研究[J]. 植物病理学报, 2010, 40(2): 210-213
KANG Zhen-hui, HUANG Jun-li. Detection of Thielaviopsis basicola in soil with real-time quantitative PCR[J]. Acta Phytopathologica Sinica, 2010, 40(2): 210-213

基金

国家自然科学基金项目(50608071)
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