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Journal of Environmental Biology

pISSN: 0254-8704 ; eISSN: 2394-0379 ; CODEN: JEBIDP

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        Abstract - Issue Jan 2019, 40 (1)                                                                                                             Back



nstantaneous and historical temperature effects on a-pinene

Development of species-specific primers for detection of Xanthomonas campestris pv. campestris causing black rot of crucifers

Paper received: 05.01.2018                          Revised received: 14.03.2018                               Accepted: 02.06.2018

 

 

Authors Info

R. Kiran1*, A. Kandan2, P. Kumar1,

D. Singh3, J. Akhtar1, B. Singh1

and S.C. Dubey1 

 

1Division of Plant Quarantine, ICAR-National Bureau of Plant Genetic Resources,

New Delhi-110 012, India

 

2Division of Insect Ecology, ICAR-National Bureau of Agricultural Insect Resources, Bengaluru-560 024, India

 

3Division of Plant Pathology, ICAR-Indian Agricultural Research Institute,

New Delhi-110 012, India 

 

 

*Corresponding Author Email :

paurajkiran@gmail.com

 

 

Abstract

Aim: Black rot of crucifers caused by Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc) is a major seed-borne disease. The present study aimed to develop a rapid diagnostic protocol for the specific and sensitive detection of this pathogen.  

 

Methodology: A specific primer set was designed based on rpf gene and optimization of PCR condition was done for specific detection of Xcc. Sensitivity of PCR for primer set was then determined by diluting the Xcc DNA and cells.      

 

Results: Specific primer set was able to amplify a specific band of 304 bp in all 11 isolates of Xcc but failed to amplify other Xanthomonas species and one each of Ralstonia solanacearum, Erwinia caratovora subsp. caratovora, Bacillus subtilis, Pseudomonas fluorescens and P. aeruginosa. The primer set was highly sensitive as it was able to detect 10 pg μl-1 bacterial DNA and up to 3x103 CFU ml-1 corresponding to 12 viable cells of Xcc which were used as template for PCR reaction         

 

Interpretation: The results suggest that developed PCR primers are highly specific and sensitive and it can be used to detect the pathogen at an early stage of infection for disease management.

 

Key words: Black rot, Diagnostic protocol, pv. campestris, rpf gene, Xanthomonas campestris

 

 

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