Detection of black rot disease causing pathogen Xanthomonas campestris pv. campestris by bio-PCR from seeds and plant parts of cole crops
Xanthomonas campestris pv. campestris which causes black rot disease of cole crops, is a serious worldwide problem, resulting in > 50% loss of crops under environmental conditions that are favourable to the pathogen. For detection of the bacteria from seeds and planting materials, two primers (Dhrp_Xcc_F: 5'-GTGGCCATGTCGTCGACTC-3' and Dhrp Xcc R: 5'-GGAATAAACTGTTTCCCCAATG-3') were designed to amplify a 769 bp DNA fragment of X. campestris pv. campestris. The primer was highly sensitive and detected 0.1ng μl-1 bacterial DNA. 136 black rot disease-infected samples of cauliflower and cabbage plants were collected from different states of India to validate the primer and 94.1% samples showed a positive reaction through bio-PCR. To detect X. campestris pv. campestris from seeds, cauliflower seeds were artificially contaminated with the bacteria at 100, 75, 50, 25, 10, 5, 1.0, 0.1 and 0.01% by dipping in broth culture containing 109 cfu ml-1 for 2.5 or 12 hours. Bio-PCR using the primers successfully detected X. campestris pv. campestris down to 0.01% contaminated seeds, whereas in NSCAA agar medium, the pathogenic bacteria was detected only at 10.0 and 1.0% contaminated seeds for 2.5- and 12-hour durations of dipping seeds in broth culture, respectively. Bio-PCR was highly sensitive to detect X. campestris pv. campestris and to rapidly diagnose the disease with great accuracy.
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Document Type: Research Article
Publication date: April 1, 2014
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