The success of Acidovorax citrulli detection by seed health testing of commercial cucurbitaceous seeds depends on the efficiency of pathogen extraction. In the present study, different extraction techniques were compared to identify factors that influence A. citrulli
detection in naturally contaminated cucurbit seeds. Embryo-infected watermelon seeds, produced by pistil inoculation, were mixed with non-infected seeds to generate samples (n = 1000 seeds) with low infection levels (approximately 104 cells per sample). Additionally, two
naturally infested melon seed lots were tested. A. citrulli was extracted from seeds by soaking or crushing, followed by one or two centrifugation steps. Samples extracted by soaking seeds yielded better amplification efficiency (103%) compared with crushing (93%), as determined by
quantitative real-time PCR analysis. This was most likely due to a reduction of the concentration of inhibitors present in the DNA samples. PCR assays using three different A. citrulli-specific primer sets highlighted that soaking followed by two centrifugation steps enhanced pathogen
detection (100% of the samples) and the mean cycle threshold (Ct) value was significantly lower than those observed for the other pathogen extraction techniques. These results indicate that the optimised extraction protocol combined with PCR analysis can improve routine seed health testing
for A. citrulli.
No Supplementary Data.
No Article Media
BACTERIAL FRUIT BLOTCH;
PATHOGEN EXTRACTION TECHNIQUES;
REAL-TIME PCR ASSAYS;
SEED HEALTH TESTING;
Document Type: Research Article
April 1, 2018
This article was made available online on December 19, 2017 as a Fast Track article with title: "Factors influencing the detection of Acidovorax citrulli in naturally contaminated cucurbitaceous seeds by PCR-based assays".
More about this publication?
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.
- Editorial Board
- Information for Authors
- Subscribe to this Title
- Membership Information
- Ingenta Connect is not responsible for the content or availability of external websites