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Progress towards a commercial PCR-based seed assay for Acidovorax avenae subsp. citrulli

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To develop a commercial-scale PCR-based assay for Acidovorax avenae subsp. citrulli in watermelon seed, parameters for immunomagnetic separation (IMS) were optimized. Optimal conditions for target cell recovery included 40 μg of polyclonal anti-AAC antibody per 108 immunomagnetic beads (IMBs) for coating, ca. 2.5 × 107 IMBs per sample and immunocapture for one hour at 4°C. These parameters consistently facilitated the recovery of A. avenae subsp. citrulli cells from suspensions containing ca.10 CFU/ml. Using lots contaminated with artificially infested seeds (104 A. avenae subsp. citrulli CFU/seed), IMS-PCR detected the pathogen in 25% and 87.5% of samples (n=10,000 seeds) with 0.01 and 0.1% infestation, respectively. For seedlots with similar infestation levels, the detection frequencies for the seedling grow-out assay (SGO) were 12.5% and 37.5%; however, the difference in detection frequency between SGO and IMS-PCR was not statistically significant. While liquid enrichment improved the detection sensitivity of IMS-PCR 100-fold, the difference in detection frequency between enrichment IMS-PCR and IMS-PCR was not statistically significant for seed samples contaminated with artificially and naturally infested seeds. These results suggest that IMS-PCR has great potential as an effective alternative to SGO for the detection of A. avenae subsp. citrulli in commercial watermelon seedlots.
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Document Type: Research Article

Publication date: April 1, 2006

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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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