First Report of Pectobacterium punjabense Causing Potato Soft Rot and Blackleg in Serbia
2021; American Phytopathological Society; Volume: 106; Issue: 5 Linguagem: Inglês
10.1094/pdis-06-21-1199-pdn
ISSN1943-7692
AutoresMarta Loc, Dragana Milošević, Maja Ignjatov, Žarko Ivanović, Dragana Budakov, Jovana Grahovac, Vanja Vlajkov, Ivana Pajčin, Mila Grahovac,
Tópico(s)Plant Pathogens and Fungal Diseases
ResumoHomePlant DiseaseVol. 106, No. 5First Report of Pectobacterium punjabense Causing Potato Soft Rot and Blackleg in Serbia PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Pectobacterium punjabense Causing Potato Soft Rot and Blackleg in SerbiaM. Loc, D. Miloševic, M. Ignjatov, Ž. Ivanović, D. Budakov, J. Grahovac, V. Vlajkov, I. Pajčin, and M. GrahovacM. Loc†Corresponding author: M. Loc; E-mail Address: [email protected]https://orcid.org/0000-0003-0273-8474University of Novi Sad, Faculty of Agriculture, Department of Plant and Environmental Protection, Novi Sad, Serbia, D. MiloševicInstitute of Field and Vegetable Crops, Laboratory for Seed Testing, Novi Sad, Serbia, M. IgnjatovInstitute of Field and Vegetable Crops, Laboratory for Seed Testing, Novi Sad, Serbia, Ž. Ivanovićhttps://orcid.org/0000-0002-4132-1367Institute for Plant Protection and Environment, Department of Plant Disease, Belgrade, Serbia, D. BudakovUniversity of Novi Sad, Faculty of Agriculture, Department of Plant and Environmental Protection, Novi Sad, Serbia, J. GrahovacUniversity of Novi Sad, Faculty of Technology, Department of Biotechnology and Pharmaceutical Engineering, Novi Sad, Serbia, V. VlajkovUniversity of Novi Sad, Faculty of Technology, Department of Biotechnology and Pharmaceutical Engineering, Novi Sad, Serbia, I. PajčinUniversity of Novi Sad, Faculty of Technology, Department of Biotechnology and Pharmaceutical Engineering, Novi Sad, Serbia, and M. GrahovacUniversity of Novi Sad, Faculty of Agriculture, Department of Plant and Environmental Protection, Novi Sad, SerbiaAffiliationsAuthors and Affiliations M. Loc1 † D. Miloševic2 M. Ignjatov2 Ž. Ivanović3 D. Budakov1 J. Grahovac4 V. Vlajkov4 I. Pajčin4 M. Grahovac1 1University of Novi Sad, Faculty of Agriculture, Department of Plant and Environmental Protection, Novi Sad, Serbia 2Institute of Field and Vegetable Crops, Laboratory for Seed Testing, Novi Sad, Serbia 3Institute for Plant Protection and Environment, Department of Plant Disease, Belgrade, Serbia 4University of Novi Sad, Faculty of Technology, Department of Biotechnology and Pharmaceutical Engineering, Novi Sad, Serbia Published Online:29 Mar 2022https://doi.org/10.1094/PDIS-06-21-1199-PDNAboutSectionsPDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Soft rot and blackleg are common diseases of potato (Solanum tuberosum) in Serbia. Opportunistic pectinolytic plant pathogens in the genus Pectobacterium cause soft rot and wilt diseases by cell wall degradation. They cause major economic losses in potato crops worldwide and are among the top 10 plant pathogenic bacteria (Mansfield et al. 2012). Potato plants ('VR808') with symptoms of wilting, slow growth, stem blackening, and tuber softening were collected from a commercial field in Zobnatica, Serbia, in July 2019 and analyzed. All symptoms occurred in the same field, and ∼5% of plants were symptomatic. Isolation was performed from 10 randomly chosen symptomatic plants and tubers. Plant tissue was surface disinfected, and 1-cm sections from the margins of lesions were macerated in sterile distilled water (SDW) for 25 min and streaked on nutrient agar. After 48 h of incubation at 26°C, predominant shiny, cream-colored, round colonies were obtained from all samples. Three representative isolates (MMZKVR1, MMZCVR2, and MMZKVR3) from independent samples were selected randomly for biochemical and pathogenicity tests. Isolates were gram-negative, nonfluorescent facultative anaerobes exhibiting pectinolytic activity on potato tuber slices and hypersensitive response on tobacco leaves. They expressed catalase activity but did not express oxidase or acid phosphatase activity or produce indole. All strains grew at 37°C in 5% NaCl and reduced nitrate. Pathogenicity was tested on healthy 3-week-old potato plants (VR808 and 'Kiebitz') grown in Baltic Tray Substrate (Hawita) in a greenhouse and tubers of the same varieties. Three stems per isolate were inoculated by toothpick piercing (Duarte et al. 2004) using a bacterial suspension (∼1 × 108 CFU/ml) and incubated under plastic bags in a greenhouse at 25 ± 2°C. Blackleg symptoms and stem wilting developed within 48 h. No symptoms were observed on plants inoculated with sterile toothpicks dipped in SDW. The pathogen was reisolated from symptomatic plants, fulfilling Koch's postulates, and sequencing of 16S rDNA confirmed the original pathogen. Three tubers per isolate were inoculated by toothpicks dipped in bacterial suspension (∼1 × 108 CFU/ml) and placed in a sealed plastic container at 25 ± 2°C. Treatment with SDW was used as a control. Tissue softening around the inoculation point developed within 48 h; no symptoms developed on controls. For molecular analyses, total DNA of isolates was extracted with a DNeasy Plant Mini Kit (Qiagen). The isolates were not detected in diagnostic PCR assays using specific primers Br1F/L1R for the detection of P. brasiliense (Duarte et al. 2004) and primers EXPCCF/EXPCCR for P. carotovorum subsp. carotovorum (Kang et al. 2003). The 16S rDNA PCR amplification was done using the universal PCR primer pair 27F/1492R (Fredriksson et al. 2013) followed by Sanger sequencing (Macrogen Europe BV). BLASTn analysis of sequences (GenBank nos. MZ048661, MZ048662, and MZ157274) revealed 100% query coverage and 100% identity to P. punjabense sequences in NCBI (MT242589 and CP038498) isolated from potato in China and Pakistan, respectively (Sarfraz et al. 2018). All three obtained isolates were proposed to belong to P. punjabense sp. nov. To further validate identification, MMZCVR2 was used for multilocus sequence analyses of five housekeeping genes: gyrA, recA, recN, rpoA, and rpoS. The gyrA (MZ161817), recA (MZ161818), recN (MZ161819), rpoA (MZ161820), and rpoS (MZ161821) sequence analysis had the highest nucleotide identity (99.44 to 100%) with P. punjabense strain SS95 (Sarfraz et al. 2018) in the NCBI GenBank database. To our knowledge, this is the first report of blackleg and soft rot caused by P. punjabense on potato in Serbia. P. punjabense causes soft rot and blackleg disease in potatoes (Sarfraz et al. 2018). Its distribution is poorly known but important because soft rot bacteria are easily transported long distances in latently infected seed tubers and can cause major economic losses.The author(s) declare no conflict of interest.References:Duarte, V., et al. 2004. J. Appl. Microbiol. 96:535. https://doi.org/10.1111/j.1365-2672.2004.02173.x Crossref, ISI, Google ScholarFredriksson, N. J., et al. 2013. PLoS One 8:e76431. https://doi.org/10.1371/journal.pone.0076431 Crossref, Google ScholarKang, H. W., et al. 2003. Plant Pathol. 52:127. https://doi.org/10.1046/j.1365-3059.2003.00822.x Crossref, ISI, Google ScholarMansfield, J., et al. 2012. Mol. Plant Pathol. 13:614. https://doi.org/10.1111/j.1364-3703.2012.00804.x Crossref, ISI, Google ScholarSarfraz, S., et al. 2018. J. Syst. Evol. 68:3551. https://doi.org/10.1099/ijsem.0.003029 Google ScholarThe author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 106, No. 5 May 2022SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 28 Apr 2022Published: 29 Mar 2022First Look: 13 Oct 2021Accepted: 12 Oct 2021 Page: 1513 Information© 2022 The American Phytopathological SocietyKeywordsblacklegpathogen detectionPectobacteriumPectobacterium punjabensepotatosoft rotThe author(s) declare no conflict of interest.PDF downloadCited byControlling soft rot of postharvest chilli pepper (Capsicum annuum L.) by an antagonist Bacillus amyloliquefaciens S917: Efficacy and action modeBiological Control, Vol. 178Diversity of Bacterial Soft Rot-Causing Pectobacterium Species Affecting Cabbage in Serbia29 January 2023 | Microorganisms, Vol. 11, No. 2Isolation and identification of Epicoccum nigrum as the causal agent of brown spot disease in Solanum tuberosum in China10 January 2023 | Plant Pathology, Vol. 49Genetic Diversity of Pectobacterium spp. on Potato in Serbia15 September 2022 | Microorganisms, Vol. 10, No. 9Antagonist Temperature Variation Affects the Photosynthetic Parameters and Secondary Metabolites of Ocimum basilicum L. and Salvia officinalis L.8 July 2022 | Plants, Vol. 11, No. 14
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