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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (05): 791-799.doi: 10.3724/SP.J.1006.2012.00791

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Quantification of Fusarium oxysporum f. sp. phaseoli Detected by Real-time Quantitative PCR in Different Common Beans Cultivars

XUE Ren-Feng1, ZHU Zhen-Dong1,*, HUANG Yan1,2,WANG Xiao-Ming1,WANG Lan-Fen1,WANG Shu-Min1,*   

  1. 1 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement,Beijing 100081, China; 2 College of Life Science and Technology, Hebei Normal University of Science & Technology, Changli 066600, China
  • Received:2011-11-14 Revised:2012-01-19 Online:2012-05-12 Published:2012-03-05

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Abstract: Fusarium wilt, caused by Fusarium oxysporum f. sp. phaseoli, is one of the most devastating vascular diseases of common bean. A efficient way to prevent and control the disease is to use resistant cultivars, so it is necessary to develop a rapid accurate method for detecting the pathogens. In this study, we developed a real-time quantitative polymerase chain reaction (qRT-PCR) protocol that quantified F. oxysporum f. sp. phaseoli DNA to a minimum of 1 pg in the plant roots and stems. Moreover, the qRT-PCR protocol asymptomatically distinguished resistant level of different bean cultivars challenged by the wilt pathogen FOP-DM01 at 6 d post inoculation. The result indicated FOP-DM01 DNA quantifications in the roots and stems of susceptible BRB-130 and A0640-1 were significantly higher than those in resistant 260205 and Heiyundou, which absolutely matched with the phenotypic identification. The use of this protocol for fast, reliable, and cost-effective quantification of F. oxysporum f. sp. phaseoli in asymptomatic tissues at early stages of the infection process is of great value for common bean breeding and studies of phytopathology and epidemiology.

Key words: Common bean, Fusarium wilt, Real time-PCR, Quantitative analysis

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