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Detection of black rot disease causing pathogen Xanthomonas campestris pv. campestris by bio-PCR from seeds and plant parts of cole crops

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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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  • Seed Science and Technology (SST) is one of the leading international journals featuring original papers and review articles on seed quality and physiology as related to seed production, harvest, processing, sampling, storage, distribution and testing. This widely recognised journal is designed to meet the needs of researchers, advisers and all those involved in the improvement and technical control of seed quality.
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