Abstract
Pythium root rot is one of the most destructive diseases on lisianthus in Japan. Current management strategies against Pythium root rot are insufficient in controlling the disease especially on hydroponically grown lisianthus. This study aimed to explore the potential of dark septate endophytic (DSE) fungi to suppress the root rot disease caused by Pythium irregulare. Among the 10 DSE screened for their pathogenicity on lisianthus, seven DSE produced a positive effect on shoot growth and were thus selected for the first screening in an axenic dual-plate assay. Accordingly, the isolate J1PC1 displayed consistent disease suppression and was thus selected as a final candidate for evaluation in small- and large-scale hydroponic experiments as well as a pot experiment using commercial soil. Shoot disease severity and disease incidence were significantly reduced by J1PC1 treatment compared with that in the untreated control. The control efficacy of J1PC1 reached 50–70%, consequently producing highly marketable lisianthus plants. J1PC1 showed no signs of direct antagonism nor parasitism on P. irregulare, suggesting that other mechanisms might be involved. Real-time PCR using species-specific primers successfully detected J1PC1 from the root tissues even after 3 months of plant treatment. Using cultural, morphological, and internal transcribed spacer-based molecular analysis, J1PC1 was identified as Hyaloscypha variabilis. Collectively, our results strongly indicate that J1PC1 is a promising biocontrol agent for Pythium root rot.
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References
Aegerter, B. J., Greathead, A. S., Pierce, L. E., & Davis, R. M. (2002). Mefenoxam-resistant isolates of Pythium irregulare in an ornamental greenhouse in California. Plant Disease, 86(6), 692–692. https://doi.org/10.1094/PDIS.2002.86.6.692B
Andrade-Linares, D. R., Grosch, R., Restrepo, S., Krumbein, A., & Franken, P. (2011). Effects of dark septate endophytes on tomato plant performance. Mycorrhiza, 21(5), 413–422. https://doi.org/10.1007/s00572-010-0351-1
Berthelot, C., Leyval, C., Chalot, M., & Blaudez, D. (2019). Interactions between dark septate endophytes, ectomycorrhizal fungi and root pathogens in vitro. FEMS Microbiology Letters, 366(13). https://doi.org/10.1093/femsle/fnz158
Deng, X., Song, X., Halifu, S., Yu, W., & Song, R. (2020). Effects of dark septate endophytes strain A024 on damping-off biocontrol, plant growth and the rhizosphere soil enviroment of Pinus sylvestris var. mongolica annual seedlings. Plants, 9(7), 913. https://doi.org/10.3390/plants9070913
Fehrer, J., Réblová, M., Bambasová, V., & Vohník, M. (2019). The root-symbioticRhizoscyphus ericae aggregate and Hyaloscypha (Leotiomycetes) are congeneric: Phylogenetic and experimental evidence. Studies in Mycology, 92, 195–225. https://doi.org/10.1016/j.simyco.2018.10.004
Feng, W., Nukaya, A., Satou, M., Fukuta, N., Ishiguro, Y., Suga, H., & Kageyama, K. (2018). Use of LAMP detection to identify potential contamination sources of plant-pathogenicPythium species in hydroponic culture systems of tomato and eustoma. Plant Disease, 102(7), 1357–1364. https://doi.org/10.1094/PDIS-10-17-1679-RE
Hambleton, S., & Sigler, L. (2005). Meliniomyces, a new anamorph genus for root-associated fungi with phylogenetic affinities to Rhizoscyphus ericae (≡ Hymenoscyphus ericae), Leotiomycetes. Studies in Mycology, 53, 1–27. https://doi.org/10.3114/sim.53.1.1
Harbaugh, B. K. (2007). Lisianthus. In N. O. Anderson (Ed.), Flower breeding and genetics (pp. 644–663). Springer. https://doi.org/10.1007/978-1-4020-4428-1_24
Harsonowati, W., Marian, M., Surono, M., & Narisawa, K. (2020). The effectiveness of a dark septate endophytic fungus, Cladophialophora chaetospira SK51, to mitigate strawberry fusarium wilt disease and with growth promotion activities. Frontiers in Microbiology, 11. https://doi.org/10.3389/fmicb.2020.00585
Huzar-Novakowiski, J., & Dorrance, A. E. (2018). Genetic diversity and population structure of Pythium irregulare from soybean and corn production fields in Ohio. Plant Disease, 102(10), 1989–2000. https://doi.org/10.1094/PDIS-11-17-1725-RE
Jumpponen, A. (2001). Dark septate endophytes - are they mycorrhizal? Mycorrhiza, 11, 207–211. https://doi.org/10.1007/s005720100112
Khastini, R. O., Ogawara, T., Sato, Y., & Narisawa, K. (2014). Control of fusarium wilt in melon by the fungal endophyte, Cadophora sp. European Journal of Plant Pathology, 139, 333–342. https://doi.org/10.1007/s10658-014-0389-6
Knapp, D. G., Németh, J. B., Barry, K., Hainaut, M., Henrissat, B., Johnson, J., Kuo, A., Lim, J. H. P., Lipzen, A., Nolan, M., Ohm, R. A., Tamás, L., Grigoriev, I. V., Spatafora, J. W., Nagy, L. G., & Kovács, G. M. (2018). Comparative genomics provides insights into the lifestyle and reveals functional heterogeneity of dark septate endophytic fungi. Scientific Reports, 8, 1–13. https://doi.org/10.1038/s41598-018-24686-4
Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35(6), 1547–1549. https://doi.org/10.1093/molbev/msy096
Lahlali, R., McGregor, L., Song, T., Gossen, B. D., Narisawa, K., & Peng, G. (2014). Heteroconium chaetospira induces resistance to clubroot via upregulation of host genes involved in jasmonic acid, ethylene, and auxin biosynthesis. PLoS One, 9(4), 1–9. https://doi.org/10.1371/journal.pone.0094144
MAFF (2018). MAFF (Ministry of Agriculture, Forestry and Fishery, Government of Japan), statistics for agriculture, forestry and fisheries. Annual wholesale market of fruits and vegetables. Retrieved September 30, 2020, from http://www.maff.go.jp/j/tokei/kouhyou/seika_orosi/index.html#y
Mandyam, K., & Jumpponen, A. (2005). Seeking the elusive function of the root-colonising dark septate endophytic fungi. Studies in Mycology, 53, 173–189. https://doi.org/10.3114/sim.53.1.173
Mao, W., Carroll, R. B., & Whittington, D. P. (1998). Association of Phoma terrestris, Pythium irregulare, and fusarium acuminatum in causing red root rot of corn. Plant Disease, 82(3), 337–342. https://doi.org/10.1094/PDIS.1998.82.3.337
McGovern, R. J. (2018). Diseases of Lisianthus. In R. McGovern & W. Elmer (Eds.), Handbook of Florists' crops diseases. Handbook of plant disease management (pp. 583–632). Springer. https://doi.org/10.1007/978-3-319-39670-5_20
Miyake, N., Nagai, H., Kato, S., Matsusaki, M., Ishikawa, H., & Kageyama, K. (2015). Detection of damping-off of cape gooseberry caused by Pythium aphanidermatum and its suppression with phosphonate. Journal of General Plant Pathology, 81, 192–200. https://doi.org/10.1007/s10327-015-0582-2
MOE (2020). MOE (Ministry of Environment, Government of Japan), Conservation of water, soil, ground, and marine environment. Retrieved September 30, 2020 from https://www.env.go.jp/water/sui-kaitei/kijun.html
Morita, Y., & Tojo, M. (2007). Modifications of PARP medium using fluazinam, miconazole, and nystatin for detection of Pythium spp. in soil. Plant Disease, 91(12), 1591–1599. https://doi.org/10.1094/PDIS-91-12-1591
Morita, S., Azuma, M., Aoba, T., Satou, H., Narisawa, K., & Hashiba, T. (2003). Induced systemic resistance of Chinese cabbage to bacterial leaf spot and Alternaria leaf spot by the root endophytic fungus, Heteroconium chaetospira. Journal of General Plant Pathology, 69, 71–75. https://doi.org/10.1007/s10327-002-0005-z
Narisawa, K., Kawamata, H., Currah, R. S., & Hashiba, T. (2002). Suppression of Verticillium wilt in eggplant by some fungal root endophytes. European Journal of Plant Pathology, 108, 103–109. https://doi.org/10.1023/A:1015080311041
Narisawa, K., Usuki, F., & Hashiba, T. (2004). Control of Verticillium yellows in Chinese cabbage by the dark septate endophytic fungus LtVB3. Phytopathology, 94(5), 412–418. https://doi.org/10.1094/PHYTO.2004.94.5.412
Newsham, K. K. (2011). A meta-analysis of plant responses to dark septate root endophytes. New Phytologist, 190(3), 783–793. https://doi.org/10.1111/j.1469-8137.2010.03611.x
Ohtaka, N., & Narisawa, K. (2008). Molecular characterization and endophytic nature of the root-associated fungus Meliniomyces variabilis (LtVB3). Journal of General Plant Pathology, 74, 24–31. https://doi.org/10.1007/s10327-007-0046-4
Onozaki, T., Satou, M., Azuma, M., Kawabe, M., Kawakatsu, K., & Fukuta, N. (2020). Evaluation of 29 lisianthus cultivars (Eustoma grandiflorum) and one inbred line of E. exaltatum for resistance to two isolates of fusarium solani by using hydroponic equipment. The Horticulture Journal, 89(4), 473–480. https://doi.org/10.2503/hortj.UTD-151
Rodriguez, R. J., White, J. F., Arnold, A. E., & Redman, R. S. (2009). Fungal endophytes: Diversity and functional roles: Tansley review. New Phytologist, 182(2), 314–330. https://doi.org/10.1111/j.1469-8137.2009.02773.x
Saeedi, R., Etemadi, N., Nikbakht, A., Khoshgoftarmanesh, A. H., & Sabzalian, M. R. (2015). Calcium chelated with amino acids improves quality and postharvest life of lisianthus (Eustoma Grandiflorum cv. cinderella lime). HortScience, 50(9), 1394–1398. https://doi.org/10.21273/hortsci.50.9.1394
Sambo, P., Nicoletto, C., Giro, A., Pii, Y., Valentinuzzi, F., Mimmo, T., Lugli, P., Orzes, G., Mazzetto, F., Astolfi, S., Terzano, R., & Cesco, S. (2019). Hydroponic solutions for soilless production systems: Issues and opportunities in a smart agriculture perspective. Frontiers in Plant Science, 10. https://doi.org/10.3389/fpls.2019.00923
Satou, M., & Fukuta, N. (2016). Control of root rot of Russell prairie gentian (Eustoma grandiflorum) in hydroponic culture using a mixture of azoxystrobin and metalaxyl M (in Japanese with English summary). Japanese Journal of Phytopathology, 82(2), 93–100. https://doi.org/10.3186/jjphytopath.82.93
Satou, M., & Fukuta, N. (2019). Control of root rot of Russell prairie gentian (Eustoma grandiflorum) caused by fusarium solani in hydroponic culture using a mixture of benomyl (in Japanese with English summary). Japanese Journal of Phytopathology, 85(1), 18–24. https://doi.org/10.3186/jjphytopath.85.18
Su, Z. Z., Mao, L. J., Li, N., Feng, X. X., Yuan, Z. L., Wang, L. W., Lin, F. C., & Zhang, C. L. (2013). Evidence for biotrophic lifestyle and biocontrol potential of dark septate endophyte Harpophora oryzae to Rice blast disease. PLoS One, 8(4), 1–14. https://doi.org/10.1371/journal.pone.0061332
Surono, & Narisawa, K. (2018). The inhibitory role of dark septate endophytic fungus Phialocephala fortinii against fusarium disease on the Asparagus officinalis growth in organic source conditions. Biological Control, 121, 159–167. https://doi.org/10.1016/j.biocontrol.2018.02.017
Sutton, J. C., Sopher, C. R., Owen-Going, T. N., Liu, W., Grodzinski, B., Hall, J. C., & Benchimol, R. L. (2006). Etiology and epidemiology of Pythium root rot in hydroponic crops: Current knowledge and perspectives. Summa Phytopathology, 32(4), 307–321. https://doi.org/10.1590/s0100-54052006000400001
Tellenbach, C., & Sieber, T. N. (2012). Do colonization by dark septate endophytes and elevated temperature affect pathogenicity of oomycetes? FEMS Microbiology Ecology, 82(1), 157–168. https://doi.org/10.1111/j.1574-6941.2012.01415.x
Tellenbach, C., Sumarah, M. W., Grünig, C. R., & Miller, J. D. (2013). Inhibition of Phytophthora species by secondary metabolites produced by the dark septate endophyte Phialocephala europaea. Fungal Ecology, 6, 12–18. https://doi.org/10.1016/j.funeco.2012.10.003
Terhonen, E., Sipari, N., & Asiegbu, F. O. (2016). Inhibition of phytopathogens by fungal root endophytes of Norway spruce. Biological Control, 99, 53–63. https://doi.org/10.1016/j.biocontrol.2016.04.006
Uematsu, S., Shiraishi, T., Shoji, T., Sekiyama, K., & Nakamura, Y. (1993). Occurrence of Pythium root rot of Eustoma grandiflorum caused by Pythium irregulare and P. spinosum in Kanto District and its chemical control (in Japanese with English summary). Proceedings of the Kanto-Tosan Plant Protection Society, 40, 167–170.
Usui, E., Takashima, Y., & Narisawa, K. (2016). Cladophialophora inabaensis sp. Nov., a new species among the dark septate endophytes from a secondary forest in Tottori, Japan. Microbes and Environments, 31(3), 357–360. https://doi.org/10.1264/jsme2.ME16016
Watanabe, H., Kageyama, K., Taguchi, Y., Horinouchi, H., & Hyakumachi, M. (2008). Bait method to detect Pythium species that grow at high temperatures in hydroponic solutions. Journal of General Plant Pathology, 74, 417–424. https://doi.org/10.1007/s10327-008-0116-2
Weiland, J. E., Santamaria, L., & Grünwald, N. J. (2014). Sensitivity of Pythium irregulare, P. sylvaticum, and P. ultimum from forest nurseries to mefenoxam and fosetyl-Al, and control of Pythium damping-off. Plant Disease, 98(7), 937–942. https://doi.org/10.1094/PDIS-09-13-0998-RE
Yakti, W., Kovács, G. M., & Franken, P. (2019). Differential interaction of the dark septate endophyte Cadophora sp. and fungal pathogens in vitro and in planta. FEMS Microbiology Ecology, 95(12), 1–12. https://doi.org/10.1093/femsec/fiz164
Acknowledgements
We are grateful to the Agribusiness Division, Toyama Kankyo Seibi Co., Ltd., for kindly providing the lisianthus seeds and seedlings. We would like to thank Ms. Makiko Fukushima and undergraduate students of Ibaraki University for their technical assistance.
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All authors contributed to the study conception and design. MM, YT, WH, and HM carried out the experiments and performed the data analysis. The first draft of the manuscript was written by MM. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Marian, M., Takashima, Y., Harsonowati, W. et al. Biocontrol of Pythium root rot on lisianthus using a new dark septate endophytic fungus Hyaloscypha variabilis J1PC1. Eur J Plant Pathol 163, 97–112 (2022). https://doi.org/10.1007/s10658-022-02459-0
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DOI: https://doi.org/10.1007/s10658-022-02459-0