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Immuno-capture PCR method for detecting Acidovorax avenae subsp. citrulli from watermelon

Published online by Cambridge University Press:  02 August 2007

Wang Xiao ,
Zhang Le ,
Xu Fu-Shou ,
Zhao Li-Han  and
Xie Guan-Lin
Wang Xiao
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
Zhang Le
Affiliation:
Chinese Academy of Inspection and Quarantine, Beijing 200025, China
Xu Fu-Shou
Affiliation:
Hangzhou Plant Protection Station, Hangzhou 310020, China
Zhao Li-Han
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
Xie Guan-Lin*
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
*
*Corresponding author. E-mail: glxie@zju.edu.cn
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Abstract

An immuno-capture polymerase chain reaction (IC-PCR) method for detection of Acidovorax avenae subsp. citrulli (AAC), the causal organism of bacterial fruit blotch (BFB) of watermelon, was developed by combining the immunosorbent enrichment (ISE) method with classical PCR and comparing with the direct PCR and growth check methods. The results showed that all A. avenae subsp. citrulli strains tested have produced 360 bp specific fragments using IC-PCR and direct PCR methods, while other strains from 10 different genera showed negative PCR results. The minimum detection concentration was about 50–100 cfu/ml and 104 cfu/ml, respectively. The IC-PCR sensitivity was 100 times higher than that of direct PCR. The examination of seven batches of different melon seeds from the markets by IC-PCR showed that one cantaloupe, two honeydew melon and two watermelon seed varieties carried the pathogen, indicating that the IC-PCR is an accurate, sensitive, rapid and low-cost technique.

Keywords

bacterial fruit blotch of watermelonPCRimmuno-capture PCR
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 4 , Issue 2 , August 2007 , pp. 173 - 179
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

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Footnotes

First published in Journal of Agricultural Biotechnology 2006, 14(6): 946–951

References

Cai, XQ, Huang, YY, Yang, JZ, Chen, J, Cai, GL and Hu, FP (2005) Pathogen identification of bacterial fruit blotch of watermelon in Fujian. Journal of Fujian Agriculture and Forestry University (Natural Science Edition) 34: 434–437 (in Chinese with English abstract).Google Scholar
Fang, ZD (1998) Research Methods of Plant Pathology. Beijing: Agricultural Press, pp. 212–218 (in Chinese).Google Scholar
Ren, YZ, Li, H and Li, GY (2004) Detection of Acidovorax avenae subsp. citrulli in melon seed using the polymerase chain reaction. Xinjiang Agricultural Sciences 41(5): 329–332 (in Chinese with English abstract).Google Scholar
Schaad, NW and Bonde, MR (1997) EIO-PCR: A highly sensitive technique for detecting seedborne fungi and bacteria (A). Seed Health Testing: Progress Towards the 21st century. Wallingford, UK: CAB International, pp. 159–164.Google Scholar
Song, WY, Sechler, AJ and Hatziloukas, E (2003) Use of PCR for rapid identification of Acidovorax avenae and A. avenae subsp. citrulli (A). Presentations from the 6th International Conference on Pseudomonas syringae Pathovars and Related Pathogens. Netherlands: Kluwer Academic, pp. 531–544.Google Scholar
van Vuurde, JWL (1997) New approach in detecting phytopathogenic bacteria by combined immunoisolation and immunoidentification assays. EPPO Bulletin 17: 139–148.CrossRefGoogle Scholar
Walcott, RD and Gitaitis, RD (2000) Detection of Acidovorax avenae subsp. citrullin in watermelon seed using immunomagnetic separation and the polymerase chain reaction. Plant Disease 84(4): 470–474.CrossRefGoogle Scholar
Xie, GL and Mew, TW (1998) A leaf inoculation method for detection of Xanthomonas oryzae pv. oryzicola from rice seed. Plant Disease 82(9): 1007–1011.Google Scholar
Zhang, X, Li, GY and Jin, Q (2002) Identification of pathogen of bacterial disease of melon and pathogen's alternate predomination investigation in field in XinJiang. Scientia Agricultura Sinica 35(7): 888–893 (in Chinese with English abstract).Google Scholar
Zhao, TC, Sun, FZ and Wang, BW (2001a) The research development of bacterial fruit blotch of watermelon. Plant Protection Technology and Extension 21: 37–38 (in Chinese).Google Scholar
Zhao, TC, Sun, FZ, Liu, SP and Kang, J (2001b) Bacterial Fruit Blotch of melon and its control. Plant Protection 27(1): 46–47 (in Chinese).Google Scholar