Morphological Characterization and Phylogeny of Pythium and Related Genera in Rayong Province, Thailand

Main Article Content

Rattasit Saelee
Kanungnid Busarakam
Prommart Koohakan*

Abstract

Most well-known microorganisms in the class Oomycetes (notably genera Phytophthora and Pythium) are pathogenic to both animals and plants due to their diverse lifestyle patterns. This study was designed to recover Pythium from composite soils (cultivated and forest soils) and water sources (fresh and brackish water) from Rayong Province. Twenty isolates of hyaline and non-septate fungal-like organisms were isolated from those sources.  The primer pair ITS4 and ITS6 were used to amplify approximately 900 bp products from Internal transcribed spacer (ITS) region and morphological characteristics including sporangium, oogonium, antheridium and oospore, were noted.  Morphological characteristics data of recovered Pythium strain can be classified into 12 source groups. ITS sequencing results revealed that eight closely related species had been recovered: Globisporangium splendens, Pythium cucurbitacearum, Pythium acanthichum, Pythium deliense, Pythium diclinum, Pythium torulosum, Phytopythium vexans and Phytopythium helicoides, which had similarities in the range 94.67-100% values at between 656 and 922 locations. Most of these species were reported as plant pathogens. Therefore, this report can be used as a guide for disease control planning.


 


Keywords: Oomycetes; identification; phylogeny; Rayong Province; Pythium


*Corresponding author: E-mail:prommart.ko@kmitl.ac.th

Article Details

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Original Research Articles

References

Schroeder, K.L., Martin, F.N., De Cock, Arthur, W.A.M., Lévesque, C.A., Spies, C.F.J., Okubara, P.A. and Paulitz, T.C., 2013.Molecular detection and quantification of Pythium species: Evolving Taxonomy, New Tools, and Challenges. Plant Disease, 97(1), 4-20.

Phillips, A.J.M., Anderson, V.L., Robertson, E.J., Secombes, C.J. and West, P.V., 2007. New insights into animal pathogenic Oomycetes. TRENDS in Microbiology, 16, 13-19.

Kamoun, S., Furzer, O., Jones, J.D.G., Judelson, H.S., Ali, G.S. , Dalio, R.J.D., Roy, S.G., Schena, L., Zambounis, A., Panabières, F., Cahill D., Ruocco, M., Figueiredo, A., Chen, X.R., Hulvey, J., Stam, R., Lamour, K., Gijzen, M., Tyler, B.M., Grünwald, N.J., Mukhtar, M. S., Tomé, D.F.A., Tör, M. , Ackerveken, G.V.D, McDowell, J., Daay, F., Fry, W.E., Kreuze, H.L., Meijer, H.J.G., Petre, B., Ristaino, J., Yoshida, K., Birch, P.R.J. and Govers, F., 2016. The top 10 Oomycete pathogens in molecular plant pathology. Molecular in Plant Pathology, 16(4), 413-434.

Tojo, M., van West, P., Hoshino, T., Kida, K., Fujii, H., Hakoda, A., Kawaguchi, Y., Mühlhauser, H.A., Van Den Berg, A.H., Küpper, F.C., Herrero, M.L., Klemsdal, S.S., Tronsmo, A.M. and Kanda H., 2012. Pythium polare, a new heterothallic oomycete causing brown discolouration of Sanioniauncinata in the Arctic and Antarctic. Fungal Biology, 116(7), 756-768.

Navi, S.S., Huynh, T., Mayers, C.G. and Yang, X.B., 2019. Diversity of Pythium spp. associated with soybean damping-off, and management implications by using foliar fungicides as seed treatments. Phytopathology Research, 1, https://doi.org/10.1186/s 42483-019-0015-9

Qiu, L., Mao, Y., Tang, L., Tang, X., and Mo, Z., 2018. Characterization of Pythium chondricola associated with red rot disease of Pyropiayezoensis (Ueda) (Bangiales, Rhodophyta) from Lianyungang, China. Journal of Oceanology and Limnology, 37, 1102-1112.

Phillips, A.J.M., Anderson, V.L., Robertson, E.J., Secombes, C.J. and West, P.V., 2007. New insights into animal pathogenic Oomycetes. Trends in Microbiology, 16, 13-19.

Bernheim, D., Dupont, D., Aptel, F., Dard, C., Chiquet, C., Normand, A.C., Piarroux, R., Cornet, M. and Maubon, D., 2019. Pythiosis: case report leading to new features in clinical and diagnostic management of this fungal-like infection. International Journal of Infectious Diseases, 86, 43-46.

Gaastra, W., Lipman, L.J.A., De Cock, A.W.A.M., Exel, T.K., Pegge, R.B.G., Scheurwater, J., Vilela, R. and Mendoza, L., 2010. Pythium insidiosum: an overview. Veterinary Microbiology, 146, 1-16.

Benhamou, N., Rey, N., Picard, K. and Tirilly, Y., 1999. Ultrastructural and cytochemical aspects of the interaction between the mycoparasite Pythium oligandrum and soilborne plant pathogens. Phytophatology, 89(6), 506-517.

Ali-Shtayeh, M.S. and Saleh, A.S.F., 1999. Isolation of Pythium acanthicum, P. oligandrum, and P. periplocum from soil and evaluation of their mycoparasitic activity and biocontrol efficacy against selected phytopathogenic Pythium species. Mycopathologia, 145, 143-153.

Deacon, J.W., 1979. Cellulose decomposition by Pythium and its relevance to substrate-group of fungi. The British Mycological Society, 77(3), 469-477.

Park, D., 1980. A two-year study of numbers of cellulolytic Pythium in river water. The British Mycological Society, 74(2), 253-258.

Warcup, J. H., 1950. The soil-plate method for isolation of fungi from soil. Nature, 166(4211), 117-118.

Masago, H., Yoshikawa, M., Fukada, M. and Nakanishi, N., 1977. Selective inhibition of Pythium spp. on a medium for direct isolation of Phytophthora spp. from soils and plants. Phytopathology, 67, 425-428.

Dhingra, O.D. and Sinclair, J.B., 1994. Basic Plant Pathology Methods. 2nd ed. Florida: CRC Press.

Stanghellini, M.E. and Kronland, W.C., 1985 Bioassay for quantification of Pythium aphanidermatum in soil. Phytopathology, 75, 1242-1245.

Abad, Z.G., Shew, H.D. and Lucas, L.T. 1994. Characterization and pathogenicity of Pythium species isolated from turfgrass with symptoms of root and crown rot in North Carolina. Phytopathology, 84, 913-921.

Van der Plaats-Niterink, A.J., 1981. Monograph of the genus Pythium. Studies in Mycology, 21, 1-242.

Ivors, K.L., 2015. Laboratory Protocols for Phytophthora Species. St. Paul: The American Phytopathological Society (APS).

Hung, P.M., Pongnak. W., Soytong, K. and Poeaim, S., 2015. Efficacy of Chaetomium species as biological control agents against Phytophthora nicotianae root rot in citrus. Mycobiology, 43(3), 288-296.

Cooke, D.E.L., Drenth, A., Duncan, J.M., Wagels, G. and Brasier, C.M., 2000. A molecular phylogeny of Phytophthora and related Oomycetes. Fungal Genetics and Biology, 30, 17-32.

Saitou, N. and Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406-425.

Felsenstein, J., 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783-791.

Uzuhashi, S., Ikeda, H., Kamekawa, A., Radmer, L. E., Kobayashi-Yamazaki, S., Kuroda, K., Funakubo, T., Ichikawa, K. And Tojo, M., 2019. Presence of two species-level groups in Globisporangium splendens isolates in Japan. European Journal of Plant Pathology, 154, 751–766.

Lévesque, C.A. and de Cock A.W.A.M., 2004. Molecular phylogeny and taxonomy of the genus Pythium. Mycological Research, 108, 1363-1383.

de Cock, A.W.A.M., Lodhi, A.M., Rintoul, T.L., Bala, K., Robideau, G.P., Abad, Z.G., Coffey, M.D., Shahzad, S. and Lévesque, C.A., 2015. Phytopythium: molecularphylogeny and systematics. Persoonia, 34, 25-39.

Nam, B. and Choi, Y.J., 2019. Phytopythium and Pythium species (Oomycota) isolated from freshwater environments of Korea. Mycobiology, 47(3), 261-272.

Ho, H.H., Chen, X.X., Zeng, H.C., and Zheng, F.C., 2012. The occurrence and distribution of Pythium species on Hainan Island of South China. Botanical Studies, 53, 525-534.

Matsiakh, I., Oszako, T., Kramarets, V. and Nowakowska, J.A., 2016. Phytophthora and Pythium species detected in rivers of the Polish-Ukrainian border areas. Baltic Forestry, 22(2), 230-238.

Ko, W.H., Wang, S.-Y. and Ann, P.-J., 2004. Pythium sukuiense, a new species from undisturbed natural forest in Taiwan. Mycologia, 96(3), 647-649.

Burr, T.J. and Stanghellini, M.E., 1973. Propagule nature and density of Pythium aphanidermatum in field soil. Phytopathology, 63, 1499-1501.