Raoultella terrigena y Pectobacterium carotovorum en hortalizas en dos provincias de Costa Rica

Autores/as

DOI:

https://doi.org/10.15517/am.v32i1.40845

Palabras clave:

pudrición blanda, fitopatología, análisis molecular

Resumen

Introducción. La pudrición blanda en los cultivos es causada por un grupo de bacterias capaces de secretar enzimas que degradan la pectina de la pared celular de las plantas y ocasionan pérdidas económicas importantes en la agricultura a nivel mundial. En Costa Rica existe poca información acerca de la distribución, hospederos y diversidad genética de los agentes causales de esta enfermedad. Objetivo. Identificar la presencia de bacterias causantes de pudrición blanda en hortalizas de la zona de Cartago y Alajuela con análisis moleculares, enzimáticos y técnicas de patogenicidad. Materiales y métodos. El estudio se llevó a cabo entre julio y octubre de 2017 en Cartago y Alajuela, Costa Rica. Se recolectaron plantas con síntomas de pudrición blanda en: chile dulce (Capcicum annum), hojas y bulbos de cebolla (Allium cepa), plantas de zucchini (Cucurbita pepo), frutos de tomate (Solanum lycopersicum) y plantas de papa (Solanum tuberosum). Se realizaron aislamientos bacterianos en un medio selectivo de cristal violeta y pectato (CVP). Se efectuaron ensayos de reacción hipersensible (RH), los aislamientos se inocularon en hojas de chile dulce y se llevaron a cabo pruebas de patogenicidad (postulados de Koch) para las bacterias positivas. Los aislamientos se identificaron molecularmente mediante el gen ARN-Ribosomal 16S. Resultados. Se aislaron cinco bacterias con actividad pectinolítica: Pt1-A, 6-M2, Ech2A, CfspA y Cfsab. Según el análisis de patogenicidad, las cepas CfspA, Cfsab y Ech2A fueron causantes de pudrición blanda en chile. No se logró reproducir los síntomas en cebolla y papa. De acuerdo con la identificación molecular, las cepas CfspA y Cfsab se clasificaron dentro del clado de Pectobacterium carotovorum, mientras que Ech2A se clasificó como Raoultella terrigena. Conclusión. Las bacterias Raoultella terrigena y Pectobacterium carotovorum se consideraron los agentes causales de pudrición blanda del chile en las zonas de Cartago y Alajuela, respectivamente.

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Citas

Agrios, G. N. (2005). Plant Pathology (5th ed.). Elsevier.

Arauz, L. (2011). Fitopatología: Un enfoque agroecológico (5ª ed.). Editorial Universidad de Costa Rica.

Asai, S., & Shirasu, K. (2015). Plant cells under siege: Plant immune system versus pathogen effectors. Current Opinion in Plant Biology, 28, 1–8. https://doi.org/10.1016/j.pbi.2015.08.008

Barny, M. A. (1995). Erwinia amylovora hrpN mutants, blocked in harpin synthesis, express a reduced virulence on host plants and elicit variable hypersensitive reactions on tobacco. European Journal of Plant Pathology, 101(3), 333–340. https://doi.org/10.1007/BF01874789

Bdliya, B. S., & Langerfeld, E. (2005). A semi-selective medium for detection, isolation and enumeration of Erwinia carotovora ssp. Carotovora from plant materials and soil. Tropical Science, 45(2), 90–96. https://doi.org/10.1002/ts.56

Bhat, K., Masood, S., Bhat, N., Bhat, M. A., Razvi, S., Mir, M., Akhtar, S., Wani, N., & Habib, M. (2010). Current status of post harvest soft rot in vegetables: A review. Asian Journal of Plant Sciences, 9(4), 200–208. https://doi.org/10.3923/ajps.2010.200.208

Bonas, U., Schulte, R., Fenselau, S., Minsavage, G. V., Staskawicz, B. J., & Stall, R. E. (1991). Isolation of a gene cluster from Xanthomonas campestris pv. Vesicatoria that determines pathogenicity and the hypersensitive response on pepper and tomato. Molecular Plant-Microbe Interactions, 4(1), 81–88. https://doi.org/10.1094/mpmi-4-081

Brisse, S., Grimont, F., & Grimont, P. A. D. (2006). The genus Klebsiella. In M. Dworkin, S. Falkow, E. Rosenberg, K.-H. Schleifer, & E. Stackebrandt (Eds.), The Prokaryotes: Volume 6: Proteobacteria: Gamma Subclass (pp. 159–196). Springer New York. https://doi.org/10.1007/0-387-30746-X_8

Brown, C., & Seidler, R. J. (1973). Potential pathogens in the environment: Klebsiella pneumoniae, a taxonomic and ecological enigma. Applied Microbiology, 25(6), 900–904. https://doi.org/10.1128/AEM.25.6.900-904.1973

Byrd, A. L., & Segre, J. A. (2016). Adapting Koch’s postulates. Science, 351(6270), 224–226. https://doi.org/10.1126/science.aad6753

Chandrashekar, B. S., Prasannakumar, M. K., Puneeth, M. E., Teli, K., Priyanka, K., Mahesh, H. B., & Desai, R. U. (2018). First report of bacterial soft rot of carrot caused by Klebsiella variicola in India. New Disease Reports, 37, Article 21. https://doi.org/10.5197/j.2044-0588.2018.037.021

Charkowski, A.O. (2007). The soft rot Erwinia. In S. S. Gnanamanickam (Ed.), Plant-Associated Bacteria (pp. 423-505). Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-4538-7_13

Charkowski, A. O. (2018). The Changing Face of Bacterial Soft-Rot Diseases. Annual Review of Phytopathology, 56(1), 269–288. https://doi.org/10.1146/annurev-phyto-080417-045906

Charkowski, A., Blanco, C., Condemine, G., Expert, D., Franza, T., Hayes, C., Hugouvieux-Cotte-Pattat, N., Solanilla, E. L., Low, D., Moleleki, L., Pirhonen, M., Pitman, A., Perna, N., Reverchon, S., Rodríguez Palenzuela, P., San Francisco, M., Toth, I., Tsuyumu, S., van der Waals, J., Yedidia, I. (2012). The role of secretion systems and small molecules in soft-rot enterobacteriaceae pathogenicity. Annual Review of Phytopathology, 50(1), 425–449. https://doi.org/10.1146/annurev-phyto-081211-173013

Charkowski, A. O., Lind, J., & Rubio-Salazar, I. (2014). Genomics of plant-associated bacteria: The soft rot enterobacteriaceae. In D. C. Gross, A. Lichens-Park, & C. Kole, (Eds.), Genomics of plant-associated bacteria (pp. 37–58). Springer. https://doi.org/10.1007/978-3-642-55378-3_2

Chinchilla, C., Gonzales, L., & Morales, F. (1979). Purdrición bacteriana del cogollo de la piña en Costa Rica. Agronomía Costarricense, 3(2), 183–185.

Choi, O., & Kim, J. (2013). Pectobacterium carotovorum subsp. Brasiliense Causing Soft Rot on Paprika in Korea. Journal of Phytopathology, 161(2), 125–127. https://doi.org/10.1111/jph.12022

Czajkowski, R., Pérombelon, M. C. M., Jafra, S., Lojkowska, E., Potrykus, M., van der Wolf, J. M., & Sledz, W. (2015). Detection, identification and differentiation of Pectobacterium and Dickeya species causing potato blackleg and tuber soft rot: A review. Annals of Applied Biology, 166(1), 18–38. https://doi.org/10.1111/aab.12166

Davidsson, P. R., Kariola, T., Niemi, O., & Palva, T. (2013). Pathogenicity of and plant immunity to soft rot pectobacteria. Frontiers in Plant Science, 4, Article 191. https://doi.org/10.3389/fpls.2013.00191

Duarte, V., Boer, S. H. D., Ward, L. J., & de Oliveira, A. M. R. (2004). Characterization of atypical Erwinia carotovora strains causing blackleg of potato in Brazil. Journal of Applied Microbiology, 96(3), 535–545. https://doi.org/10.1111/j.1365-2672.2004.02173.x

Estrada, R. S. G., Reyes, C. J., Fasio, J. A. C., Molar, R. A., Zequera, I. M., & Rangel, M. D. M. (2000). Marchitez bacteriana en chile bell causada por Erwinia carotovora subsp carotovora. Revista Mexicana de Fitopatología, 18(2), 120–124.

Etalo, D. W., Stulemeijer, I. J. E., Esse, H. P. van, de Vos, R. C. H., Bouwmeester, H. J., & Joosten, M. H. A. J. (2013). System-Wide Hypersensitive Response-Associated Transcriptome and Metabolome Reprogramming in Tomato. Plant Physiology, 162(3), 1599–1617. https://doi.org/10.1104/pp.113.217471

Fan, H. C., Zeng, L., Yang, P. W., Guo, Z. X., & Bai, T. T. (2016). First report of banana soft rot caused by Klebsiella variicola in China. Plant Disease, 100(2), 517–517. https://doi.org/10.1094/PDIS-05-15-0586-PDN

Feng, F., & Zhou, J. M. (2012). Plant–bacterial pathogen interactions mediated by type III effectors. Current Opinion in Plant Biology, 15(4), 469–476. https://doi.org/10.1016/j.pbi.2012.03.004

Fontecha, G. (2003). Análisis comparativo de las comunidades de procariotas intestinales de Rothschildia lebeau (Lepidoptera) mediante los polimorfismos en los fragmentos terminales de restricción (T-RFLP) del gen ARNr 16S [Tesis de Maestría no publicada]. Universidad de Costa Rica.

Gardan, L., Gouy, C., Christen, R., & Samson, R. (2003). Elevation of three subspecies of Pectobacterium carotovorum to species level: Pectobacterium atrosepticum sp. nov., Pectobacterium betavasculorum sp. nov. and Pectobacterium wasabiae sp. nov. International Journal of Systematic and Evolutionary Microbiology, 53(2), 381–391. https://doi.org/10.1099/ijs.0.02423-0

Gillis, A., Santana, M. A., Rodríguez, M., & Romay, G. (2017). First Report of Bell Pepper Soft-Rot Caused by Pectobacterium carotovorum subsp. Brasiliense in Venezuela. Plant Disease, 101(9), 1671–1671. https://doi.org/10.1094/PDIS-03-17-0361-PDN

Govrin, E. M., & Levine, A. (2000). The hypersensitive response facilitates plant infection by the necrotrophic pathogen Botrytis cinerea. Current Biology, 10(13), 751–757. https://doi.org/10.1016/S0960-9822(00)00560-1

Gupta, R., Lee, S. E., Agrawal, G. K., Rakwal, R., Park, S., Wang, Y., & Kim, S. T. (2015). Understanding the plant-pathogen interactions in the context of proteomics-generated apoplastic proteins inventory. Frontiers in Plant Science, 6, Article 352. https://doi.org/10.3389/fpls.2015.00352

Hall, T. A. (1999). BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98. https://doi.org/10.14601/Phytopathol_Mediterr-14998u1.29

Hélias, V., Hamon, P., Huchet, E., Wolf, J. V. D., & Andrivon, D. (2012). Two new effective semiselective crystal violet pectate media for isolation of Pectobacterium and Dickeya. Plant Pathology, 61(2), 339–345. https://doi.org/10.1111/j.1365-3059.2011.02508.x

Herrera, J., & González, L. (1977). Development and control of potato blackleg, caused by Erwinia carotovora var. Atroseptica, in Costa Rica. Agronomía Costarricense, 1(2), 161–163.

Huang, H. E., Ger, M. J., Yip, M. K., Chen, C. Y., Pandey, A. K., & Feng, T. Y. (2004). A hypersensitive response was induced by virulent bacteria in transgenic tobacco plants overexpressing a plant ferredoxin-like protein (PFLP). Physiological and Molecular Plant Pathology, 64(2), 103–110. https://doi.org/10.1016/j.pmpp.2004.05.005

Hugouvieux-Cotte-Pattat, N., Condemine, G., & Shevchik, V. E. (2014). Bacterial pectate lyases, structural and functional diversity. Environmental Microbiology Reports, 6(5), 427–440. https://doi.org/10.1111/1758-2229.12166

Izard, D., Ferragut, C., Gavini, F., Kersters, K., De Ley, J., & Leclerc, H. (1981). Klebsiella terrigena, a New Species from Soil and Water. International Journal of Systematic Bacteriology, 31(2), 116–127. https://doi.org/10.1099/00207713-31-2-116

Jaramillo, A., Huertas, C. A., & Gómez, E. D. (2016). First Report of Bacterial Stem Rot of Tomatoes Caused by Pectobacterium carotovorum subsp. Brasiliense in Colombia. Plant Disease, 101(5), 830. https://doi.org/10.1094/PDIS-08-16-1184-PDN

Jensen, M. A., Webster, J. A., & Straus, N. (1993). Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Applied and Environmental Microbiology, 59(4), 945–952. https://doi.org/10.1128/AEM.59.4.945-952.1993

Kim, J. H., Joen, Y. H., Kim, S. G., & Kim, Y. H. (2007). First report on bacterial soft rot of graft-cactus Chamaecereus silvestrii caused by Pectobacterium carotovorum subsp. Carotovorum in Korea. The Plant Pathology Journal, 23(4), 314-317. https://doi.org/10.5423/PPJ.2007.23.4.314

Kim, H. S., Thammarat, P., Lommel, S. A., Hogan, C. S., & Charkowski, A. O. (2011). Pectobacterium carotovorum Elicits Plant Cell Death with DspE/F but the P. carotovorum DspE Does Not Suppress Callose or Induce Expression of Plant Genes Early in Plant–Microbe Interactions. Molecular Plant-Microbe Interactions, 24(7), 773–786. https://doi.org/10.1094/MPMI-06-10-0143

Klement, Z. (1963). Rapid detection of the pathogenicity of phytopathogenic pseudomonads. Nature, 199, 299–300. https://doi.org/10.1038/199299b0

Klement, Z., & Goodman, R. (1967). The hypersensitive reaction to infection by bacterial plant pathogens. Annual Review of Phytopathology, 5(1), 17-44. https://doi.org/10.1146/annurev.py.05.090167.000313

Knittel, M. D., Seidler, R. J., Eby, C., & Cabe, L. M. (1977). Colonization of the botanical environment by Klebsiella isolates of pathogenic origin. Applied and Environmental Microbiology, 34(5), 557–563. https://doi.org/10.1128/AEM.34.5.557-563.1977

Kumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33(7), 1870–1874. https://doi.org/10.1093/molbev/msw054

Kushalappa, A. C., Yogendra, K. N., & Karre, S. (2016). Plant innate immune response: qualitative and quantitative resistance. Critical Reviews in Plant Sciences, 35(1), 38–55. https://doi.org/10.1080/07352689.2016.1148980

Lane, D. (1991). 16S/23S rRNA sequencing. In E. Stackebrandt, & M. Goodfellow (Eds.), Nucleic acid techniques in bacterial systematics (pp. 115-175). John Wiley & Sons, Inc.

Ma, B., Hibbing, M. E., Kim, H. S., Reedy, R. M., Yedidia, I., Breuer, J., Breuer, J., Glasner, J. D., Perna, N. T., & Kelman, A. (2007). Host range and molecular phylogenies of the soft rot enterobacterial genera Pectobacterium and Dickeya. Phytopathology, 97(9), 1150–1163. https://doi.org/10.1094/PHYTO-97-9-1150

McFarland, J. (1907). The nephelometer: An instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. Journal of the American Medical Association, 49(14), 1176–1178. https://doi.org/10.1001/jama.1907.25320140022001f

Meng, X., Chai, A., Shi, Y., Xie, X., Ma, Z., & Li, B. (2016). Emergence of bacterial soft rot in cucumber caused by Pectobacterium carotovorum subsp. Brasiliense in China. Plant Disease, 101(2), 279-287. https://doi.org/10.1094/PDIS-05-16-0763-RE

Nabhan, S., Boer, S. H. D., Maiss, E., & Wydra, K. (2012). Taxonomic relatedness between Pectobacterium carotovorum subsp. Carotovorum, Pectobacterium carotovorum subsp. Odoriferum and Pectobacterium carotovorum subsp. Brasiliense subsp. Nov. Journal of Applied Microbiology, 113(4), 904–913. https://doi.org/10.1111/j.1365-2672.2012.05383.x

Ngadze, E., Coutinho, T. A., & van der Waals, J. E. (2010). First Report of Soft Rot of Potatoes Caused by Dickeya dadantii in Zimbabwe. Plant Disease, 94(10), 1263–1263. https://doi.org/10.1094/PDIS-05-10-0361

Nguyen-the, C., & Carlin, F. (1994). The microbiology of minimally processed fresh fruits and vegetables. Critical Reviews in Food Science & Nutrition, 34(4), 371–401. https://doi.org/10.1080/10408399409527668

Perombelon, M. C., & Kelman, A. (1980). Ecology of the soft-rot Erwinias. Annual Review of Phytopathology, 18(1), 361–387. https://doi.org/10.1146/annurev.py.18.090180.002045

Pitman, A. R., Harrow, S. A., & Visnovsky, S. B. (2010). Genetic characterisation of Pectobacterium wasabiae causing soft rot disease of potato in New Zealand. European Journal of Plant Pathology, 126(3), 423–435. https://doi.org/10.1007/s10658-009-9551-y

Pitman, A. R., Wright, P. J., Galbraith, M. D., & Harrow, S. A. (2008). Biochemical and genetic diversity of pectolytic enterobacteria causing soft rot disease of potatoes in New Zealand. Australasian Plant Pathology, 37(6), 559–568. https://doi.org/10.1071/AP08056

Pu, X. M., Zhou, J. N., Lin, B. R., & Shen, H. F. (2012). First report of bacterial foot rot of rice caused by a Dickeya zeae in China. Plant Disease, 96(12), 1818–1818. https://doi.org/10.1094/PDIS-03-12-0315-PDN

Ritchie, D. (2000). Bacterial spot of pepper and tomato. The Plant Health Instructor. The American Phytopathological Society. https://doi.org/10.1094. PHI-I-2000-1027-01.

Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D. L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M. A., & Huelsenbeck, J. P. (2012). MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space. Systematic Biology, 61(3), 539–542. https://doi.org/10.1093/sysbio/sys029

Sánchez, K. R., Zúñiga, O. C., Meneses, M. B., & González, A. Q. (2019). Etiología de las pudriciones en el tallo de Hylocereus costaricensis, provocadas por Enterobacter hormaechei, en Costa Rica. Agronomía Costarricense, 43(2), 61–73. https://doi.org/10.15517/rac.v43i2.37949

Schaad, N. W., Jones, J. B., & Chun, W. (2001). Laboratory guide for the identification of plant pathogenic bacteria. American Phytopathological Society Press.

Singh, J., & Kaur, L. (2016). Advances in potato chemistry and technology (2nd ed.). Academic press.

Staats, M., van Baarlen, P., & van Kan, J. A. L. (2005). Molecular Phylogeny of the Plant Pathogenic Genus Botrytis and the Evolution of Host Specificity. Molecular Biology and Evolution, 22(2), 333–346. https://doi.org/10.1093/molbev/msi020

Toth, I. K., Bell, K. S., Holeva, M. C., & Birch, P. R. J. (2003). Soft rot Erwiniae: From genes to genomes. Molecular Plant Pathology, 4(1), 17–30. https://doi.org/10.1046/j.1364-3703.2003.00149.x

Umesha, S., Richardson, P. A., Kong, P., & Hong, C. X. (2008). A novel indicator plant to test the hypersensitivity of phytopathogenic bacteria. Journal of Microbiological Methods, 72(1), 95–97. https://doi.org/10.1016/j.mimet.2007.11.002

van-der-Merwe, J. J., Coutinho, T. A., Korsten, L., & van der Waals, J. E. (2010). Pectobacterium carotovorum subsp. Brasiliensis causing blackleg on potatoes in South Africa. European Journal of Plant Pathology, 126(2), 175–185. https://doi.org/10.1007/s10658-009-9531-2

Volcy, C. (2008). Genesis and evolution of Koch postulates and their relationship with phytopathology. A review. Agronomía Colombiana, 26(1), 107-115.

Waleron, M., Waleron, K., & Lojkowska, E. (2015). First Report of Pectobacterium carotovorum subsp. Brasiliense causing soft rot on potato and other vegetables in Poland. Plant Disease, 99(9), 1271–1271. https://doi.org/10.1094/PDIS-02-15-0180-PDN

Wei-Salas, S., & Durán-Quirós, A. (2015). Characterization of land use in the main agricultural areas of the Major Metropolitan Area of Costa Rica. Agronomía Costarricense, 39(1), 151–160.

Williams, B., Kabbage, M., Kim, H.-J., Britt, R., & Dickman, M. B. (2011). Tipping the balance: Sclerotinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment. PLOS Pathogens, 7(6), e1002107. https://doi.org/10.1371/journal.ppat.1002107

Wright, P., Triggs, C., & Burge, G. (2005). Control of bacterial soft rot of calla (Zantedeschia spp.) by pathogen exclusion, elimination and removal. New Zealand Journal of Crop and Horticultural Science, 33(2), 117–123. https://doi.org/10.1080/01140671.2005.9514340

Zhang, J., Shen, H., Pu, X., Lin, B., & Hu, J. (2014). Identification of Dickeya zeae as a Causal Agent of Bacterial Soft Rot in Banana in China. Plant Disease, 98(4), 436–442. https://doi.org/10.1094/PDIS-07-13-0711-RE

Publicado

2021-01-01

Cómo citar

Cubero-Agüero, D., Brenes-Guillén, L., Vidaurre -Barahona, D., & Uribe-Lorío, L. (2021). Raoultella terrigena y Pectobacterium carotovorum en hortalizas en dos provincias de Costa Rica. Agronomía Mesoamericana, 32(1), 178–195. https://doi.org/10.15517/am.v32i1.40845