First Report of Soft Rot of Cabbage Caused by Pectobacterium wasabiae in Japan

HomePlant DiseaseVol. 105, No. 8First Report of Soft Rot of Cabbage Caused by Pectobacterium wasabiae in Japan PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Soft Rot of Cabbage Caused by Pectobacterium wasabiae in JapanTaketo Fujimoto, Takato Nakayama, Takehiro Ohki, and Tetsuo MaokaTaketo Fujimotohttps://orcid.org/0000-0002-8893-2325Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization (HARC/NARO), Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, JapanSearch for more papers by this author, Takato NakayamaHokkaido Agricultural Research Center, National Agriculture and Food Research Organization (HARC/NARO), Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, JapanSearch for more papers by this author, Takehiro OhkiHokkaido Agricultural Research Center, National Agriculture and Food Research Organization (HARC/NARO), Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, JapanSearch for more papers by this author, and Tetsuo Maoka†Corresponding author: T. Maoka; E-mail Address: [email protected]Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization (HARC/NARO), Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, JapanSearch for more papers by this authorAffiliationsAuthors and Affiliations Taketo Fujimoto Takato Nakayama Takehiro Ohki Tetsuo Maoka † Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization (HARC/NARO), Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, Japan Published Online:8 Sep 2021https://doi.org/10.1094/PDIS-02-21-0238-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleCabbage (Brassica oleracea var. capitata) is one of the important vegetables in Japan. In the summer of 2019, some cabbages with soft rot were found in commercial fields in Hokkaido, the northern island in Japan. All diseased plants showed gray to brown discoloration and expanding water-soaked lesions on leaves. We obtained two independent strains (NACAB191 and NACAB192) from diseased leaves. DNA from these strains yielded an expected single-size amplicon with the primer set of PhF/PhR for Pectobacterium wasabiae (De Boer et al. 2012) by PCR but did not yield the expected amplicon with the primer set of BR1f/L1r for P. carotovorum subsp. brasiliense (Duarte et al. 2004) and Eca1f/Eca2r for P. atrosepticum (De Boer and Ward 1995) by PCR. These two strains grew at 37°C and had the ability to utilize raffinose and lactose. These bacterial strains were gram-negative and rod-shaped. The bacterium was positive for O-nitrophenyl-beta-D-galactopyranoside, N-acetylglucosaminyl transferase, gelatin liquefaction, and acid production from D-galactose, lactose, melibiose, raffinose, citrate, and trehalose. The bacterium was negative for indole production and acid production from maltose, α-methyl-D-glucoside, sorbitol, D-arabitol, inositol, inulin, and melezitose. All strains exhibited pectolytic activity on potato slices. The sequence analysis of 16S rDNA (LC597897 and LC597898) showed more than 98% identities to P. wasabiae strain (e.g., HAFL01 in Switzerland) by BLAST analysis. In addition, multilocus sequence analysis (Ma et al. 2007) was performed by MEGA10 (Kumar et al. 2018) using concatenated DNA sequences of seven housekeeping genes (aconitate hydratase [acnA, LC597923 and LC597924], glyceraldehyde-3-phosphate dehydrogenase A [gapA, LC597970 and LC597971], isocitrate dehydrogenase [icdA, LC597996 and LC597997], malate dehydrogenase [mdh, LC598022 and LC598023], mannitol-1-phosphate dehydrogenase [mtlD, LC598048 and LC598049], glucose-6-phosphate isomerase [pgi, LC598074 and LC598075], and gamma-glutamyl phosphate reductase [proA, LC598079 and LC598080]), and all clustered NACAB191 and NACAB192 into a clade containing other confirmed strains of P. wasabiae. As a result, these two strains shared high identity with each other (>98%, E-values showed 0). The clade containing these two strains was consistently placed in a larger clade with the other P. wasabiae and 100% bootstrap support for its separation from other Pectobacterium species available in GenBank when the consensus tree was constructed using the maximum likelihood method. Pathogenicity of these strains against cabbage (cv. Rakuen) was confirmed by the field experiments with 5-weeks-growth plants sprayed with bacterial suspension (1 × 107 CFU/ml). Thirty cabbages per strain were used in this study (12 plants treated the suspension of NACAB191 and 16 plants treated the suspension of NACAB192), which died with the same soft rot symptoms about 4 weeks after inoculation, whereas water-inoculated plants remained symptomless. Strains reisolated from the artificially diseased stems were confirmed as P. wasabiae using the methods as biochemical characterization and multiple genetic analyses. Based on the disease symptoms and on the cultural, molecular, and pathological features of the strains, we conclude that the soft rot symptoms of cabbage in Hokkaido in 2019 were caused by P. wasabiae. To our knowledge, this is the first report of P. wasabiae as a soft rot disease agent of cabbage in Japan.The author(s) declare no conflict of interest.References:De Boer, S. H., and Ward, L. J. 1995. Phytopathology 85:854. https://doi.org/10.1094/Phyto-85-854 Crossref, ISI, Google ScholarDe Boer, S. H., et al. 2012. Phytopathology 102:937. https://doi.org/10.1094/PHYTO-04-12-0083-R Link, ISI, Google ScholarDuarte, V., et al. 2004. J. Appl. Microbiol. 96:535. https://doi.org/10.1111/j.1365-2672.2004.02173.x Crossref, ISI, Google ScholarKumar, S., et al. 2018. Mol. Biol. Evol. 35:1547. Crossref, ISI, Google ScholarMa, B., et al. 2007. Phytopathology 97:1150. https://doi.org/10.1094/PHYTO-97-9-1150 Link, ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by Research program on development of innovative technology (01022C).DetailsFiguresLiterature CitedRelated Vol. 105, No. 8 August 2021SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionComparison of banana plants in silicon-deficient soil amended or nonamended with calcium silicate and infected by Fusarium oxysporum f. sp. cubense (W. Zellner et al.). Photo credit: A. A. Fortunato and F. A. Rodrigues. Infected spicebush tree outer bark showing black vascular discoloration of the sapwood typical of laurel wilt (R. Olatinwo et al.). Photo credit: R. Olatinwo. Maize plants naturally infected by Bipolaris zeicola (S. S. Liu et al.). Photo credit: S. S. Liu. Metrics Article History Issue Date: 1 Dec 2021Published: 8 Sep 2021First Look: 6 Apr 2021Accepted: 24 Mar 2021 Page: 2236 Information© 2021 The American Phytopathological SocietyFundingResearch program on development of innovative technologyGrant/Award Number: 01022CKeywordsprokaryotescabbagesoft rotPectobacteriumThe author(s) declare no conflict of interest.PDF downloadCited byDiversity of Bacterial Soft Rot-Causing Pectobacterium Species Affecting Cabbage in Serbia29 January 2023 | Microorganisms, Vol. 11, No. 2