Abstract
We developed a method to detect Acidovorax avenae subsp. citrulli from cucurbit seeds that is more sensitive than conventional growing-out methods. Seeds were cultivated in test tubes for 2 weeks, and the bacteria grew semi-selectively on the seeds and seedlings. The plants were then vigorously shaken in water to suspend bacterial cells, and the suspension was spread on a selective medium for identifying the bacterium by colony appearance. The method is suitable for research purposes to evaluate the percentage of seeds infested with the bacteria, but not to detect bacterial contamination in commercial lots with thousands of seeds.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bahar O, Efrat M, Hadar E, Dutta B, Walcott RR, Burdman S (2008) New subspecies-specific polymerase chain reaction-based assay for the detection of Acidovorax avenae subsp. citrulli. Plant Pathol 57:754–763
Hirata K, Kobayashi Y, Mizuno A, Saito N, Sumiya T (1998) Quarantine pests not occur in Japan concerning the growing site inspection in exporting countries (in Japanese). Plant Prot 52:83–88
Hopkins DL, Thompson CM (2002) Seed transmission of Acidovorax avenae subsp. citrulli in cucurbits. HortScience 37:924–926
Horita H, Tsukamoto T, Uematsu H, Yasuoka S (2006) Occurrence of melon bacterial fruit blotch caused by Acidovorax avenae subsp. citrulli in Japan (Abstract in Japanese). Jpn J Phytopathol 72:82
Koenraadt H, Borst R, van Vliet A, Hoekstra M, van Schie J, Buimer M (2005) Detection of Acidovorax avenae subsp. citrulli with the sweatbox method (Abstract). Phytopathology 95:962
Komiya Y, Shirakawa T, Abiko K (2003) Rapid diagnosis of watermelon bacterial fruit blotch caused by Acidovorax avenae subsp. citrulli with immunological agglutination methods (in Japanese with English summary). Jpn J Phytopathol 69:15–18
Kubota M, Hagiwara N, Shirakawa T, Nishi K (2009) The method for detection of Acidovorax avenae subsp. citrulli from each melon seed (in Japanese). Annu Rep Kansai Plant Prot 51:23–25
Langston DB Jr, Walcott RD, Gitaitis RD, Sanders FH Jr (1999) First report of a fruit rot of pumpkin caused by Acidovorax avenae subsp. citrulli in Georgia (Abstract). Plant Dis 83:199
Martin HL, O’Brien RG, Abbott DV (1999) First report of Acidovorax avenae subsp. citrulli as a pathogen of cucumber (Abstract). Plant Dis 83:965
Matsuura T, Shirakawa T, Sato M, Inoue Y, Azegami K (2008) Detection and isolation of Acidovorax avenae subsp. citrulli from watermelon seeds using membrane filtration immunostaining (in Japanese with English summary). Jpn J Phytopathol 74:153–156
Ogiso H, Fujinaka M, Shimizu T (2001) Occurrence of watermelon bacterial fruit blotch in Nagano Prefecture (in Japanese). Annu Rep Kanto-Tosan Plant Prot Soc 48:33–36
Ogiso H, Fujinaga M, Matsuzaki R, Uematsu H (2005) First report of a bacterial fruit blotch of wax gourd by Acidovorax avenae subsp. citrulli, and possibility of seed transmission in wax gourd (Abstract in Japanese). Jpn J Phytopathol 71:290
Oya H, Nakagawa H, Saito N, Uematsu H, Ohara T (2008) Detection of Acidovorax avenae subsp. citrulli from seed using LAMP method (in Japanese with English summary). Jpn J Phytopathol 74:304–310
Sato M, Shirakawa T, Matsuura T, Tashiro Y (2006) A method for the detection of Acidovorax avenae subsp. citrulli in watermelon and melon seeds using enrichment system with seedlings (Abstract in Japanese). Jpn J Phytopathol 72:311–312
Shirakawa T (2002) Development of selective medium for detection and quantification (in Japanese). In: Ministry of Agriculture, Forest and Fisheries (ed) Development of methods to control bacterial fruit blotch of watermelon. Ministry of Agriculture, Forest and Fisheries, Tokyo, pp 2–13
Shirakawa T, Aizawa M, Komiya Y, Abiko K (2000a) Development of semiselective medium for isolation and detection of Acidovorax avenae subsp. citrulli from seeds and plant materials (Abstract in Japanese). Jpn J Phytopathol 66:132
Shirakawa T, Kikuchi S, Kato T, Abiko K, Kawai A (2000b) Occurrence of watermelon bacterial fruit blotch in Japan (in Japanese with English summary). Jpn J Phytopathol 66:223–231
Shirakawa T, Komiya Y, Abiko K (2003) Population dynamics of Acidovorax avenae subsp. citrulli on infested seeds and on subsequent seedlings of watermelon (in Japanese with English summary). Jpn J Phytopathol 69:102–106
Suzuki A, Sato M, Yamauchi N, Oya H, Matsuura T, Shirakawa T (2008) Improved PCR method using new primers and polyvinylpolypyrrolidone for detection of Acidovorax avenae subsp. citrulli from plant materials (Abstract in Japanese). Jpn J Phytopathol 74:42
Tomita Y, Ogawara T, Shirakawa T, Sato M, Kashima T, Nakanishi H (2006) Occurrence of bacterial fruit blotch of melon caused by Acidovorax avenae subsp. citrulli in Ibaraki Prefecture (Abstract in Japanese). Jpn J Phytopathol 72:312
Walcott RR, Gitaitis RD (2000) Detection of Acidovorax avenae subsp. citrulli in watermelon seed using immunomagnetic separation and the polymerase chain reaction. Plant Dis 84:470–474
Walcott RR, Castro AC, Fessehaie A, Ling K (2006) Progress towards a commercial PCR-based assay for Acidovorax avenae subsp. citrulli. Seed Sci Technol 34:101–116
Acknowledgments
This work was supported by ‘Research and Development Projects for Application in Promoting New Policy of Agriculture, and Forestry and Fisheries’ by the Ministry of Agriculture, Forestry and Fisheries, Japan. We are very grateful to Mr. Kazuaki Hara (Nanto Seeds Co., Ltd.) and Dr. Masatoshi Sato (National Center of Seeds and Seedlings) for their very helpful advice. We also greatly thank Mr. Katsuyuki Yamauchi, Mr. Tanehisa Bessyo, Ms. Toshiko Umezawa, Ms. Hidemi Fujimoto and Dr. Kazufumi Nishi for laboratory support.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kubota, M., Hagiwara, N. & Shirakawa, T. A method to evaluate percentage of cucurbit seeds infested with Acidovorax avenae subsp. citrulli, pathogen of bacterial fruit blotch. J Gen Plant Pathol 77, 112–115 (2011). https://doi.org/10.1007/s10327-010-0291-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10327-010-0291-9