Journal of Environmental Biology

Application of loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive detection of fungal pathogen, Colletotrichum capsici in Capsicum annuum

 Aravindaram Kandan*, Jameel Akhtar, Baleshwar Singh, Deepa Pal, Dinesh Chand,

Prakash Chandra Agarwal and Sunil Chandra Dubey

Division of Plant Quarantine, Indian Council of Agricultural Research (ICAR)-National Bureau of Plant Genetic Resources (NBPGR), Pusa Campus, New Delhi-110 012, India

*Corresponding Author E-mail: genekannz@gmail.com

Publication Data

Paper received:

26 May 2016

Revised received:

01 August 2016

Re-revised received:

12 August 2016

Accepted:

22 August 2016

Abstract

Colletotrichum capsici is an economically important pathogen that severely affects the fruits of chilli (Capsicum annuum) plants. Loop-mediated isothermal amplification (LAMP) assay was used for accurate and sensitive detection of C. capsici. Sequences of β-tubulin gene of C. capsici were used as target template to design LAMP primers. The primers were very specific to C. capsici and did not react against any other seed-borne fungi of chillies. LAMP assay was sensitive and could detect as little as 10 fg μl^-1 of C. capsici pathogen in comparison with 1 ng ?l^-1 of C. capsici detection using polymerase chain reaction. Further test detection of infected chilli seeds collected from different locations in India showed that the LAMP assay was more sensitive than the PCR-based detection of C. capsici. This LAMP assay is simple, fast and sensitive in detection of C. capsici infected field samples, tissue culture materials and for schedule screening of plant quarantine materials under exchange. In the present study, LAMP assay has been used for fast and sensitive detection of C. capsici pathogen.?????????

 Key words

Colletotrichum capsici, Capsicum annuum, Fungal pathogens, LAMP assay

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