First Report of Pectobacterium carotovorum subsp. brasiliense Causing Soft Rot on Squash and Watermelon in Serbia
2019; American Phytopathological Society; Volume: 103; Issue: 10 Linguagem: Inglês
10.1094/pdis-12-18-2213-pdn
ISSN1943-7692
AutoresNevena Zlatković, Anđelka Prokić, Katarina Gašić, Nemanja Kuzmanović, Milan Ivanović, Aleksa Obradović,
Tópico(s)Plant-Microbe Interactions and Immunity
ResumoHomePlant DiseaseVol. 103, No. 10First Report of Pectobacterium carotovorum subsp. brasiliense Causing Soft Rot on Squash and Watermelon in Serbia PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Pectobacterium carotovorum subsp. brasiliense Causing Soft Rot on Squash and Watermelon in SerbiaN. Zlatković, A. Prokić, K. Gašić, N. Kuzmanović, M. Ivanović, and A. ObradovićN. ZlatkovićUniversity of Belgrade, Faculty of Agriculture, Belgrade, Serbia, A. Prokić†Corresponding author: A. Prokić; E-mail Address: [email protected]http://orcid.org/0000-0002-6203-5669University of Belgrade, Faculty of Agriculture, Belgrade, Serbia, K. GašićInstitute for Plant Protection and Environment, Belgrade, Serbia, N. KuzmanovićJulius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Epidemiology and Pathogen Diagnostics, Braunschweig, Germany, M. IvanovićUniversity of Belgrade, Faculty of Agriculture, Belgrade, Serbia, and A. ObradovićUniversity of Belgrade, Faculty of Agriculture, Belgrade, SerbiaAffiliationsAuthors and Affiliations N. Zlatković1 A. Prokić1 † K. Gašić2 N. Kuzmanović3 M. Ivanović1 A. Obradović1 1University of Belgrade, Faculty of Agriculture, Belgrade, Serbia 2Institute for Plant Protection and Environment, Belgrade, Serbia 3Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Epidemiology and Pathogen Diagnostics, Braunschweig, Germany Published Online:25 Jul 2019https://doi.org/10.1094/PDIS-12-18-2213-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Watermelon (Citrullus lanatus) and squash (Cucurbita pepo) are traditionally grown in northern Serbia, in the province of Vojvodina. Most of the cucurbit cultivars grown in Serbia originate from imported seed. In July 2013, round, water-soaked lesions were observed on the fruits on sporadic squash plants grown in two fields of the breeder variety trials in Bački Petrovac. Affected tissue was light brown, slightly sunken, soft, and macerated. From symptomatic plants (cultivar GL Maxima), showing 30% symptom intensity, a uniform population of bacterial colonies was isolated, and one strain was chosen for the collection. In August of the following season, infection of the vine was observed on watermelon plants (cultivar unknown) grown in a commercial field in Čelarevo. Soft rot brownish lesions developed on infected stems, resulting in collapse and wilting of entire vines. The number of plants affected with symptoms was less than 10%. From the colonies isolated from the diseased tissue, two bacterial strains were purified and stored in the collection. Being pectolytic and isolated from cucurbits, the squash strain and two watermelon strains were subjected to further characterization. The strains developed white-gray, nonmucoid, round colonies with irregular margins on nutrient agar medium. They were gram and oxidase negative, tobacco HR and catalase positive, facultative anaerobes, nonfluorescent, caused soft rot on potato tuber slices, grew at 37°C, and reduced nitrate. They neither utilized arginine nor produced acid from sorbitol and inositol, showing characteristics of Pectobacterium species (De Boer and Kelman 2001). The PCR assay was carried out using primers BR1f/L1r (Duarte et al. 2004), specific for Pectobacterium carotovorum subsp. brasiliense (Pcb). A specific fragment of 322 bp was amplified for all three strains tested. The 16S rDNA sequence analysis (GenBank nos. MH031784, MH031785, and MH031786) showed 99% identity to the sequences of Pcb isolated from cucumber (CP020350), pepper (KX377597), and cabbage (KY021040) in China and potato in Japan (LC146476). The recA (MK911735, MK911736, and MK911737) and gapA (MK911738, MK911739, and MK911740) sequence analysis of three strains showed the highest nucleotide identity (100 to 98.35% and 99.42 to 98.08%, respectively) with Pcb previously deposited in NCBI GenBank database. Pathogenicity of the strains was tested on 2-week-old watermelon (cv. Rosa) and melon (cv. Sezam) seedlings, grown in a commercial potting mix in a greenhouse, as well as on mature zucchini fruits. Three seedlings per strain were inoculated by spraying with bacterial suspension (approx. 1 × 108 CFU/ml) using a handheld sprayer. Inoculated plants were incubated under plastic bags for 24 h and afterward maintained in a greenhouse at 28 ± 2°C. Two days after inoculation, water-soaked lesions developed on cotyledons of the watermelon and melon plants. No symptoms were observed on control plants sprayed with water. Three zucchini fruits were inoculated by pricking with a syringe and hypodermic needle and leaving a droplet of bacterial suspension (approx. 1 × 107 CFU/ml) at the point of inoculation. Inoculated fruits were placed on wet filter paper in a sealed plastic container at 28 ± 2°C. Treatment with sterile distilled water was used as a negative control. Discoloration of the tissue and soft rot spreading from the inoculation point developed within 48 h from inoculation. No symptoms developed on the control fruits. To complete Koch's postulates, bacteria were reisolated from the inoculated plant and fruit tissue and were identified as Pcb by PCR assay (Duarte et al. 2004). This is the first report of Pcb causing soft rot of squash and watermelon in Serbia. The origin of the pathogen remained unknown. It might have been present in Serbia for some time but misidentified in previous studies that could not discriminate this subspecies or recently introduced by infected seed.The author(s) declare no conflict of interest.References:De Boer, S. H., and Kelman, A. 2001. Page 56 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd Ed. American Phytopathological Society, St. Paul, MN. https://doi.org/10.1007/978-94-010-0003-1 Crossref, 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 ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by Ministry of Education, Science and Technological Development of Republic of Serbia (grant no. III46008).DetailsFiguresLiterature CitedRelated Vol. 103, No. 10 October 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionBell pepper cv. Bomby infected with zucchini yellow mosaic virus along with cucumber mosaic virus, pepper mild mottle virus, and tobacco mosaic virus (Verma et al.). Photo credit: S. Tripathi. Severe stunting, leaf chlorosis, and horizontal head growth of sunflower infected with Plasmopara halstedii, causal agent of downy mildew (Humann et al.). Photo credit: S. Markell. Metrics Article History Issue Date: 3 Oct 2019Published: 25 Jul 2019First Look: 22 May 2019Accepted: 17 May 2019 Pages: 2667-2667 Information© 2019 The American Phytopathological SocietyFundingMinistry of Education, Science and Technological Development of Republic of SerbiaGrant/Award Number: III46008KeywordsPectobacterium carotovorum subsp. brasiliensewatermelonsquashidentificationsoft rotThe author(s) declare no conflict of interest.Cited byDiversity of Bacterial Soft Rot-Causing Pectobacterium Species Affecting Cabbage in Serbia29 January 2023 | Microorganisms, Vol. 11, No. 2Occurrence, identification, and host range of Pectobacterium brasiliense causing soft rot on seed potato tubers in Turkey20 October 2022 | Journal of Plant Diseases and Protection, Vol. 130, No. 1Genetic Diversity of Pectobacterium spp. on Potato in Serbia15 September 2022 | Microorganisms, Vol. 10, No. 9Bakteriyel yumuşak çürüklük hastalığına neden olan Pectobacterium polaris'in konukçu aralığının belirlenmesi26 July 2022 | Mustafa Kemal Üniversitesi Tarım Bilimleri Dergisi, Vol. 27, No. 2Host range and virulence diversity of Pectobacterium carotovorum subsp. brasiliense strain RDKLR infecting radish in India, and development of a LAMP-based diagnostics1 June 2022 | Journal of Applied Microbiology, Vol. 132, No. 6Pectobacterium brasiliense (soft rot and blackleg of ornamentals and potato)CABI Compendium, Vol. CABI CompendiumComplete Genome Sequence Resource for the Necrotrophic Plant-Pathogenic Bacterium Pectobacterium carotovorum WPP14Yingyu Liu, Tyler C. Helmann, Paul Stodghill, and Melanie J. Filiatrault17 November 2020 | Plant Disease, Vol. 105, No. 1Pectobacterium brasiliense: Genomics, Host Range and Disease Management5 January 2021 | Microorganisms, Vol. 9, No. 1Pectobacterium and Dickeya: Environment to Disease Development5 January 2021
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