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• W2945626139 abstract "HomePlant DiseaseVol. 103, No. 11First Report of Dickeya dianthicola as a Causal Agent of Bacterial Soft Rot of Potato in Hawaii PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Dickeya dianthicola as a Causal Agent of Bacterial Soft Rot of Potato in HawaiiGamze Boluk and Mohammad ArifGamze BolukDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822Search for more papers by this author and Mohammad Arif†Corresponding author: M. Arif; E-mail Address: [email protected]http://orcid.org/0000-0002-5887-2050Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822Search for more papers by this authorAffiliationsAuthors and Affiliations Gamze Boluk Mohammad Arif † Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822 Published Online:23 Aug 2019https://doi.org/10.1094/PDIS-11-18-2094-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Bacterial soft rot caused by Dickeya sp. is one of the most economically important diseases of potato (Solanum tuberosum) around the world. Infected potato plants show wilting, slow growth, and wet, oozy, black stems (Czajkowski et al. 2011). Twenty-one infected plants, showing symptoms of wilting and black stem extending upward, were collected from a potato field at Oahu, Hawaii, in January 2018. Bacteria were isolated on crystal violet pectate medium and incubated at 26 to 28°C for 48 to 78 h. Isolated colonies were streaked on tetrazolium chloride medium (Ahmed et al. 2018) to obtain single colonies. Genomic DNA from four strains was extracted and purified using the UltraClean Microbial DNA Isolation Kit (Mo Bio, Carlsbad, CA) following the manufacturer’s instructions. The Dickeya dnaA (chromosomal replication initiator protein, DnaA) housekeeping gene was used to design a primer set, dnaA-F (5′-TAACAACGTGAACCCCAAGCA-3′) and dnaA-R (5′-TCTTCTTTGATGTCGTGACTTTC-3′), to amplify the partial dnaA gene region of Dickeya species for Sanger sequencing. The PCR reaction contained 10 μl of GoTaq Green Master Mix (Promega, Madison, WI), 1 μl of 5 mM of each forward and reverse primer, 1 μl of template DNA, and 7 μl of Ultra-Pure DNase/RNase-Free distilled water. The dnaA PCR conditions were initial denaturation at 94°C for 5 min followed by 35 cycles of denaturation at 94°C for 20 s each, annealing at 58°C for 60 s, extension at 72°C for 60 s, and final extension at 72°C for 2 min. PCR was performed in a T100 Thermal cycler (Bio-Rad, Hercules, CA). Agarose gel (1.5%) electrophoresis was used to separate PCR amplicons; gels were stained with 0.5 μg/ml of ethidium bromide. The bands were visualized under a UV illuminator. Amplified PCR products were cleaned by adding 2 µl of ExoSAP-IT (Affymetrix, Santa Clara, CA) in 5 µl of PCR product and incubated at 37°C for 15 min followed by 80°C for another 15 min. Treated templates were sequenced using both dnaA-F and dnaA-R primers at GENEWIZ (La Jolla, CA). Obtained sequences of each strain were aligned and edited manually for high accuracy. The consensus sequences of each strain were checked with BLASTn against the NCBI GenBank nucleotide and genome databases to confirm identity. The BLASTn outcomes demonstrated 100% query coverage and 100% identity only with D. dianthicola sequences. The sequences were submitted to GenBank under the accession numbers MK189268 (PL25), MK189269 (PL22), MK189270 (PL23), and MK189271 (PL24). Pectinolytic activity of each strain was assessed by surface inoculation of a potato tuber slice using the stab inoculation method; toothpicks were used to pick and inoculate the bacterial colonies. Negative control potato tuber slices were inoculated with sterile water. The experiment was performed in three replicates, and all inoculated tuber slices were incubated at 28°C. Pectinolytic activity (maceration) was observed after 24 h. No pectinolytic activity was observed in the negative controls. Pathogenicity assays were performed with healthy potato plants grown in 20-cm-diameter plastic pots under the temperature-controlled greenhouse. The plants were inoculated using single and pure bacterial colonies at three consecutive nodes starting from the base of the stem using a sterile scalpel and incubated at 25 to 28°C; five strains were used for inoculation. The experiment was performed three times. Typical disease symptoms, including blackleg on the stem, were observed within 3 to 4 days. Strains were reisolated from artificially inoculated plants and confirmed as D. dianthicola. Based on disease symptoms and cultural, molecular, and pathological features of the strains, we concluded that the disease was caused by D. dianthicola. This is the first report of D. dianthicola causing soft rot of potato in Hawaii. This incident warrants further surveys for pectinolytic bacteria causing blackleg-like symptoms on potato and other vegetable crops in Hawaii.The author(s) declare no conflict of interest.References:Ahmed, F. A., et al. 2018. Sci. Rep. 8:15972. https://doi.org/10.1038/s41598-018-34275-0 Crossref, ISI, Google ScholarCzajkowski, R., et al. 2011. Plant Pathol. 60:999. https://doi.org/10.1111/j.1365-3059.2011.02470.x Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: This work was supported by the USDA National Institute of Food and Agriculture, Hatch project 9038H, managed by the College of Tropical Agriculture and Human Resources.DetailsFiguresLiterature CitedRelated Vol. 103, No. 11 November 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionAdvanced symptoms of bacterial blotch disease on mushroom caps (Osdaghi et al.). Photo credit: C. Bull. Powdery mildew caused by Golovinomyces neosalviae on Salvia fruticosa (Soylu et al.). Photo credit: S. Soylu. Metrics Article History Issue Date: 4 Nov 2019Published: 23 Aug 2019First Look: 22 May 2019Accepted: 17 May 2019 Pages: 2943-2943 Information© 2019 The American Phytopathological SocietyFundingUSDA National Institute of Food and AgricultureGrant/Award Number: Hatch project 9038HKeywordsDickeya dianthicolapotatosoft rotplant bacteriablacklegThe author(s) declare no conflict of interest.Cited byDickeya dianthicola (slow wilt of Dianthus and potato)CABI Compendium, Vol. CABI CompendiumBioactive potential of Albizia lebbeck extract against phytopathogens and protective properties on tomato plant against speck disease in greenhousePhysiological and Molecular Plant Pathology, Vol. 117Species of Dickeya and Pectobacterium Isolated during an Outbreak of Blackleg and Soft Rot of Potato in Northeastern and North Central United States14 August 2021 | Microorganisms, Vol. 9, No. 8Complete Genome Sequence Resource for the Necrotrophic Plant-Pathogenic Bacterium Dickeya dianthicola 67-19 Isolated From New Guinea ImpatiensYingyu Liu, Tyler C. Helmann, Paul Stodghill, and Melanie J. Filiatrault9 March 2021 | Plant Disease, Vol. 105, No. 4First Report of Pectobacterium brasiliense Causing Soft Rot on Brassica oleracea var. sabellica in Hawaii, United StatesGamze Boluk, Dario Arizala, Jordie Ocenar, Julie Mokwele, Joshua Silva, Shefali Dobhal, Jensen Uyeda, Anne M. Alvarez, and Mohammad Arif18 August 2020 | Plant Disease, Vol. 104, No. 10First Report of Pectobacterium brasiliense Causing Bacterial Soft Rot and Blackleg Diseases of Potato in HawaiiDario Arizala, Shefali Dobhal, Sujan Paudel, Gamze Boluk, Joshua Silva, Amjad A. Ahmad, Jensen Uyeda, Jari Sugano, Anne M. Alvarez, and Mohammad Arif22 July 2020 | Plant Disease, Vol. 104, No. 9Genome-Informed Recombinase Polymerase Amplification Assay Coupled with a Lateral Flow Device for In-Field Detection of Dickeya SpeciesGamze Boluk, Shefali Dobhal, Alex B. Crockford, Michael Melzer, Anne M. Alvarez, and Mohammad Arif12 June 2020 | Plant Disease, Vol. 104, No. 8Comparative genomics reveals signature regions used to develop a robust and sensitive multiplex TaqMan real‐time qPCR assay to detect the genus Dickeya and Dickeya dianthicola12 February 2020 | Journal of Applied Microbiology, Vol. 128, No. 6First Report of Bacterial Soft Rot and Blackleg on Potato Caused by Pectobacterium parmentieri in HawaiiDario Arizala, Shefali Dobhal, Sujan Paudel, Samudra Gunarathne, Gamze Boluk, and Mohammad Arif5 January 2020 | Plant Disease, Vol. 104, No. 3" @default.
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