First Report of Pectobacterium brasiliense Causing Soft Rot on Brassica oleracea var. sabellica in Hawaii, United States
2020; American Phytopathological Society; Volume: 104; Issue: 10 Linguagem: Inglês
10.1094/pdis-04-20-0701-pdn
ISSN1943-7692
AutoresGamze Bölük, Dario Arizala, Jordie R. Ocenar, Julie Mokwele, Joshua Silva, Shefali Dobhal, Jensen Uyeda, Anne M. Alvarez, Mohammad Arif,
Tópico(s)Plant-Microbe Interactions and Immunity
ResumoHomePlant DiseaseVol. 104, No. 10First Report of Pectobacterium brasiliense Causing Soft Rot on Brassica oleracea var. sabellica in Hawaii, United States PreviousNext DISEASE NOTES OPENOpen Access licenseFirst 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 ArifGamze BolukDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Dario ArizalaDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Jordie OcenarDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Julie MokweleDepartment of Agriculture, Forestry and Fisheries, Arcadia, Pretoria, South Africa, Joshua SilvaDepartment of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Shefali DobhalDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Jensen UyedaDepartment of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., Anne M. AlvarezDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A., and Mohammad Arif†Corresponding author: M. Arif; E-mail Address: arif@hawaii.eduhttp://orcid.org/0000-0002-5887-2050Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A.AffiliationsAuthors and Affiliations Gamze Boluk1 Dario Arizala1 Jordie Ocenar1 Julie Mokwele2 Joshua Silva3 Shefali Dobhal1 Jensen Uyeda3 Anne M. Alvarez1 Mohammad Arif1 † 1Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A. 2Department of Agriculture, Forestry and Fisheries, Arcadia, Pretoria, South Africa 3Department of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, U.S.A. Published Online:18 Aug 2020https://doi.org/10.1094/PDIS-04-20-0701-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Kale (Brassica oleracea var. sabellica L.), family Brassicaceae, is an economically important vegetable crop and an excellent source of natural antioxidants, consumed in many countries worldwide (Ligor et al. 2013). In June 2019, 11 kale plants (variety: Starbor), showing symptoms of wilting and soft rot on the stem, were collected from a field on Oahu, Hawaii. An estimated 30 to 40% of total plants were infected. Small pieces of stems were taken (5 × 5 mm), surface sterilized with 0.6% sodium hypochlorite for 1 min, and rinsed three times with sterilized water. Later, the tissues were macerated in a sterile 1.5-ml centrifuge tube containing 100 μl of sterile water; macerated tissues (∼10 μl) were streaked onto crystal violet pectate (CVP) agar medium and incubated at 26 to 28°C for 48 to 78 h (Dobhal et al. 2020; Helias et al. 2012). Pectolytic bacteria that created pits on CVP were restreaked on nutrient agar plates containing tetrazolium chloride to obtain pure single colonies. Genomic DNA from 11 bacterial isolates was extracted using the DNeasy Blood and Tissue Kit (Qiagen) following the manufacturer's instructions. The dnaA, gapA, and gyrB housekeeping genes were used for identity determination and phylogenetic analysis. For amplification, the dnaA primer set PCR conditions and components, as described by Dobhal et al. (2020), were used for all three primer sets (dnaA gene primers from Dobhal et al. 2020; gapA gene: gapA-F1, 5′-ATATGACTATCAAAGTAGGTATCAACGG-3′, and gapA-R2, 5′-CGTACCAGGAAACCAGTTTCAC-3′; and gyrB gene: gyrB-F1, 5′-ATGTCGAATTCTTATGACTCCTCA-3′, and gyrB-R1, 5′-TGGCTTTCTTGCTGTAGCCTTC-3′). The amplification reaction contained 10 μl of GoTaq Green Master Mix (Promega), 1 μl (5 mM) of each forward and reverse primer, 1 μl of template DNA, and 7 μl of distilled water. The PCR conditions, PCR product cleanup, and sequencing protocols were followed as described by Dobhal et al. (2020). Using the error-free consensus sequences, BLASTn results showed 100% query coverage and ∼98% identity with Pectobacterium brasiliense sequences. All the sequences were submitted to the NCBI GenBank database under the following accession numbers: for dnaA gene, MN544620 to MN544630 (all 11 strains); gapA gene, MN732539 and MN732540 (PL131, PL141); and gyrB gene, MN732542 and MN732543 (PL131, PL141). Pathogenicity assays were performed with three strains on kale plants grown in plastic pots under the temperature-controlled greenhouse. Bacterial inoculum (200 µl of 3.4 × 108 CFU/ml) was inoculated into three consecutive nodes using sterile pipet tips. Controls were inoculated with sterile water; the experiment was performed in three replicates. Typical disease symptoms, wilting across the plant and soft rot on the stem, were observed after 24 h, whereas no symptoms were observed on control plants. To confirm successful P. brasiliense infection, strains were reisolated from inoculated plants and confirmed as P. brasiliense through sequencing. In the phylogenetic tree, the kale strains were grouped with seven other P. brasiliense strains retrieved from the NCBI GenBank database. Pectobacterium and Dickeya species, causing soft rot, have been reported in Hawaii on potato but not on kale (Arizala et al. 2020; Boluk and Arif 2019). Only in Brazil, P. brasiliense has been reported to cause soft rot on kale (Queiroz et al. 2017). So far, this is the first report of P. brasiliense causing bacterial soft rot on kale in Hawaii and the United States.The author(s) declare no conflict of interest.References:Arizala, D., et al. 2020. Plant Dis. 104:970. https://doi.org/10.1094/PDIS-09-19-1894-PDN Link, Google ScholarBoluk, G., and Arif, M. 2019. Plant Dis. 103:2943. https://doi.org/10.1094/PDIS-11-18-2094-PDN Link, ISI, Google ScholarDobhal, S., et al. 2020. J. Appl. Microbiol. 128:1703. https://doi.org/10.1111/jam.14579 Crossref, ISI, Google ScholarHelias, V., et al. 2012. Plant Pathol. 61:339. https://doi.org/10.1111/j.1365-3059.2011.02508.x Crossref, ISI, Google ScholarLigor, M., et al. 2013. Food Anal. Methods. 6:630. https://doi.org/10.1007/s12161-012-9367-9 Crossref, ISI, Google ScholarQueiroz, M. F., et al. 2017. Plant Dis. 101:2144. https://doi.org/10.1094/PDIS-02-17-0266-PDN Link, 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. 104, No. 10 October 2020SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionSymptoms of yellow leaf disease of Areca catechu caused by areca palm velarivirus 1 (H. X. Wang et al.). Photo credit: X. Huang. Fungal fruiting bodies of Phyllachora maydis on corn foliage resemble spots of tar (J. Valle-Torres et al.). Photo credit: C. Cruz. Geranium (Pelargonium hortorum) showing pale green and little leaves, phyllody, virescence, and witches'-broom (A. R. Amirmijani et al.). Photo credit: M. Azadvar. Metrics Article History Issue Date: 25 Sep 2020Published: 18 Aug 2020First Look: 8 May 2020Accepted: 4 May 2020 Page: 2721 Information© 2020 The American Phytopathological SocietyFundingUSDA National Institute of Food and AgricultureGrant/Award Number: Hatch project 9038HKeywordsPectobacterium brasiliensekalebacterial soft rotplant bacteriaThe author(s) declare no conflict of interest.Cited byDevelopment of a multiplex TaqMan qPCR targeting unique genomic regions for the specific and sensitive detection of Pectobacterium species and P. parmentieri1 April 2022 | Journal of Applied Microbiology, Vol. 132, No. 4Editorial: Genome-Wide Analyses of Pectobacterium and Dickeya Species3 March 2022 | Frontiers in Plant Science, Vol. 13First Report of Pectobacterium brasiliense Causing Soft Rot on Mizuna (Brassica rapa var. japonica) in the United StatesDiksha Klair, Joshua Silva, Dario Arizala, Gamze Boluk, Shefali Dobhal, Amjad A. Ahmad, Jensen Uyeda, Anne M. 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