DOI QR코드

DOI QR Code

Discovery of a new primer set for detection and quantification of Ilyonectria mors-panacis in soils for ginseng cultivation

  • Farh, Mohamed El-Agamy (Graduate School of Biotechnology and Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Han, Jeong A. (Gyeonggi-do Agricultural Research & Extension Services) ;
  • Kim, Yeon-Ju (Graduate School of Biotechnology and Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Kim, Jae Chun (Department of Oriental Medicinal Biotechnology, College of Life Sciences, Kyung Hee University) ;
  • Singh, Priyanka (Department of Oriental Medicinal Biotechnology, College of Life Sciences, Kyung Hee University) ;
  • Yang, Deok-Chun (Graduate School of Biotechnology and Ginseng Bank, College of Life Science, Kyung Hee University)
  • Received : 2016.10.14
  • Accepted : 2017.07.20
  • Published : 2019.01.15

Abstract

Background: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. Methods: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. Results: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples ($R^2=0.95$), disease severity index ($R^2=0.99$), and colony-forming units ($R^2=0.87$). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of $5.82{\pm}2.35pg/g$ to $892.34{\pm}103.70pg/g$ of soil. Conclusion: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

Keywords

References

  1. Kim YJ, Jeon JN, Jang MG, Oh JY, Kwon WS, Jung SK, Yang DC. Ginsenoside profiles and related gene expression during foliation in Panax ginseng Meyer. J Ginseng Res 2013;38:66-72. https://doi.org/10.1016/j.jgr.2013.11.001
  2. Leung KW, Wong AS. Pharmacology of ginsenosides: a literature review. Chin Med 2010;5:20. https://doi.org/10.1186/1749-8546-5-20
  3. Siraj FM, Natarajan S, Huq MA, Kim YJ, Yang DC. Structural investigation of ginsenoside Rf with PPARg major transcriptional factor of adipogenesis and its impact on adipocyte. J Ginseng Res 2014;39:141-7. https://doi.org/10.1016/j.jgr.2014.10.002
  4. Ohh SH, Yu YH, Kim KH, Cho DH. Studies on control of soil-borne diseases and insects of ginseng and development of antifungal compound. In: Korea Ginseng and Tobacco Research Inst., Ginseng Cultivation Bull; 1992. p. 121-84.
  5. Yu YH, Ohh SH. Research on ginseng diseases in Korea. Korean J Ginseng Sci 1993;17:61-8.
  6. Ziezold M, Reeleder RD, Hall R, Proctor JTA. Effect of drenching soil with benomyl, propiconazole, and fluazinam on incidence of disappearing root rot of ginseng. J Ginseng Res 1998;22:237-43.
  7. Farh MEA, Kim YJ, Kim YJ, Yang DC. Mini review: Cylindrocarpon destructans/Ilyonectria radicicola-species complex: causative agent of ginseng root-rot disease and rusty symptoms. J Ginseng Res 2018;42:9-15. https://doi.org/10.1016/j.jgr.2017.01.004
  8. Rossman AY, Samuels GJ, Rogerson CT, Lowen R. Genera of Bionectriaceae, Hypocreaceae and Nectriaceae (Hypocreales, Ascomycetes). Stud Mycol J 1999;42:1-248.
  9. Booth C. The genus Cylindrocarpon. Myc Papers 1966;104:1-56.
  10. Sutherland JR, Shrimpton GM, Surrock RN. Diseases and insects in British Columbia forest seedling nurseries. Forestry Canada and B.C. Ministry of Forests, Victoria. FRDA Report. 1989. No. 065.
  11. Unestam T, Beyer-Ericson L, Strand M. Involvement of Cylindrocarpon destructans in root death of Pinus sylvestris seedlings: pathogenic behavior and predisposing factors. Scand J For Res 1989;4:521-35. https://doi.org/10.1080/02827588909382585
  12. Seifert KA, McMullen CR, Yee D, Reeleder RD, Dobinson KF. Molecular differentiation and detection of ginseng-adapted isolates of the root rot fungus Cylindrocarpon destructans. Phytopathology 2003;93:1533-42. https://doi.org/10.1094/PHYTO.2003.93.12.1533
  13. Hildebrand AA. Root rot of ginseng in Ontario caused by members of the genus Ramularia. Can J Res 1935;12:82-114. https://doi.org/10.1139/cjr35-007
  14. Lyr H, Kluge E. Zusammenhange zwischen Pathogenitat, Enzym-und Toxinproduktion bei Cylindricarpon radicicola. Phytopathology 1968;62:220-31. https://doi.org/10.1111/j.1439-0434.1968.tb03036.x
  15. Chaverri P, Salgado C, Hirooka Y, Rossman AY, Samuels GJ. Delimitation of Neonectria and Cylindrocarpon (Nectriaceae, Hypocreales, Ascomycota) and related generawith Cylindrocarpon like anamorphs. Stud Mycol 2011;68:57-78. https://doi.org/10.3114/sim.2011.68.03
  16. Cabral A, Rego C, NascImento T, Oliveira H, Groenewald JZ, Crous PW. Multigene analysis and morphology reveal novel Ilyonectria species associated with black foot disease of grapevines. J Fungal Biol 2011;116:62-80.
  17. Cabral A, Groenewald JZ, Rego C, Oliveira H, Crous PW. Cylindrocarpon root rot: multi-gene analysis reveals novel species within the Ilyonectria radicicolaspecies complex. Mycol Prog 2012;11:655-88. https://doi.org/10.1007/s11557-011-0777-7
  18. Lombard L, Bezuidenhout CM, Crous PW. Ilyonectria black foot rot associated with Proteaceae. Australasian Plant Pathol 2012;42:337-49. https://doi.org/10.1007/s13313-012-0188-5
  19. Pathrose B, Jones EE, Jaspers MV, Ridgway HJ. High genotypic and virulence diversity in Ilyonectria liriodendri isolates associated with black foot disease in New Zealand vineyards. Plant Pathol 2014;63:613-24. https://doi.org/10.1111/ppa.12140
  20. Kernaghan G, Reeleder RD, Hoke SMT. Quantification of Cylindrocarpon destructans f. sp. panacis in soils by real-time PCR. Plant Pathol 2007;56:508-16. https://doi.org/10.1111/j.1365-3059.2006.01559.x
  21. Steenkamp ET, Wingfield BD, Coutinho TA, Wingfield MJ, Marasas WF. Differentiation of Fusarium subglutinans f. sp. pini by histone gene sequence data. Appl Environ Microbiol 1999;65:3401-6. https://doi.org/10.1128/AEM.65.8.3401-3406.1999
  22. Mesapogu S, Babu BK, Bakshi A, Reddy SS, Saxena S, Srivastava AK, Arora DK. Rapid detection and quantification of Fusarium udum in soil and plant samples using real-time PCR. J Plant Pathol Microbiol 2011. http://dx.doi.org/10.4172/2157-7471.1000107.
  23. Farh ME, Kim YJ, Singh P, Yang DC. Cross interaction between Ilyonectria morspanacis isolates infecting Korean ginseng and ginseng saponins in correlation with their pathogenicity. Phytopathology 2017;107:561-9. https://doi.org/10.1094/PHYTO-05-16-0210-R
  24. Farh ME, Kim YCJ, Van An H, Sukweenadhi J, Singh P, Huq MA, Yang DC. Burkholderia ginsengiterrae sp. nov. and Burkholderia panaciterrae sp. nov., antagonistic bacteria against root rot pathogen Cylindrocarpon destructans, isolated from ginseng soil. Arch Microbiol 2015;197:439-47. https://doi.org/10.1007/s00203-014-1075-y
  25. Singh P, Kim YJ, Hoang VA, Farh ME, Yang DC. Sphingomonas panacis sp. nov., isolated from rhizosphere of rusty ginseng. Antonie Van Leeuwenhoek 2015;108:711-20. https://doi.org/10.1007/s10482-015-0527-y
  26. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 1997;25:4876-82. https://doi.org/10.1093/nar/25.24.4876
  27. Reeleder RD, Roy R, Capell B. Seed and root rots of ginseng (Panax quinquefolius L) caused by Cylindrocarpon destructans and Fusarium spp. J Ginseng Res 2002;26:151-8. https://doi.org/10.5142/JGR.2002.26.3.151
  28. Colony-count technique at $25^{\circ}C$ ET ISO 6611: milk and milk products enumeration of colony-forming units of yeasts and/or molds. 2012 [internet]. [cited in 2016 Aug 15]; [about 8 p.] Available from: https://law.resource.org/pub/et/ibr/et.iso. 6611.2012.pdf.
  29. Howard RJ, Garland JA, Seaman WL. Diseases and pests of vegetable crops in Canada: an illustrated compendium. Entomological Society of Canada & Canadian Phytopathological Society; 1994.

Cited by

  1. The history, etiology, and management of ginseng replant disease: a Canadian perspective in review vol.101, pp.6, 2019, https://doi.org/10.1139/cjps-2021-0106