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Identification and characterisation of causal pathogens of Pestalotiopsis leaf fall disease in Hevea brasiliensis using a detached leaf technique

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Abstract

Pestalotiopsis leaf fall disease (PLFD) has emerged as a significant and detrimental foliar disease affecting Hevea brasiliensis in Malaysia since 2017. The disease poses a severe threat to rubber plantations, leading to economic losses and impacting the country’s rubber industry. Understanding the factors contributing to lesion incidence and the potential presence of a fungal complex is crucial for effective disease management. This study aimed to address key objectives, including the identification of primary fungal pathogens responsible for lesion development and examination of the role of co-infections in lesion incidence. To achieve these objectives, five fungal isolates (Neopestalotiopsis surinamensis, Colletotrichum conoides, Lasiodiplodia theobromae, Phyllosticta fallopiae and Letendraea cordylinicola) previously identified through the Internal Transcribe Spacer (ITS) regions were subjected to in vitro artificial inoculation experiments on detached rubber leaves. Various combinations of these fungal isolates were tested to evaluate their interactions as possible causal pathogens of PLFD. Three variables were assessed, viz. type of water source (sterile distilled water and rainwater), presence of leaf wounds, and inoculation techniques (conidial suspensions and mycelial agar plugs). Lesion incidence was evaluated based on sporulating occurrences on the detached leaves, recorded seven days after inoculation. Statistical analysis, including non-parametric Pearson’s chi-square tests revealed that the fungal isolates Lasiodiplodia theobromae, Colletotrichum conoides and Neopestalotiopsis surinamensis were the primary contributors to lesion incidence, accounting for 83%, 69%, and 57% of cases, respectively. The study also revealed that lesion incidence was not significantly influenced by inoculation techniques or type of water source but was greater in wounded leaves compared to unwounded leaves. Co-inoculation experiments demonstrated increased lesion incidence, highlighting the role of co-infections in disease severity. Importantly, this study elucidated a potential fungal complex associated with PLFD and provided insights into its epidemiology. The findings contribute to deeper understanding of the disease, offering valuable guidance for effective disease management strategies in H. brasiliensis cultivation in Malaysia.

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References

  1. Adam MAZ, Murnita MM, Aizat SN, Azimah I (2018) Serangan penyakit luruhan daun Pestalotiopsis Ke Atas Pokok Getah. Buletin Sains Dan Teknologi 18:62–64

    Google Scholar 

  2. Syameel M, Irwan AS, Latif D, Supian SMS, Azham Z, Kamaruzaman Y, Adam MAZ, Aizat SN (2020) Plot pemerhatian penyakit luruhan daun Pestalotiopsis Di Kampung Sg. Tekali, Hulu Langat, Selangor. Buletin Sains Dan Teknologi. 20:20–25

  3. Safiah A, Aizat SN (2022) Serangan penyakit luruhan daun Pestalotiopsis mengikut pecahan negeri (Januari - Disember 2022). Jawatankuasa Teknikal Pestalotiopsis Kebangsaan Bil. 3/2022, Kuala Lumpur, Malaysia, 16th December 2022

  4. Anon (2021) Report of the 2nd Meeting of the ANRPC Technical Committee on Plant Protection. Virtual meeting hosted by Association of Natural Rubber Producing Countries (ANRPC), Kuala Lumpur, Malaysia, 3rd February 2021. pp. 10

  5. Adam MAZ, Murnita MM, Aizat SN, Nor Afiqah M, Safiah A (2020) Factors for the spread of Pestalotiopsis fungal infection in tropics. In Pestalotiopsis Leaf Fall Disease: Identification, Prevention and Control. Malaysian Rubber Board, Kuala Lumpur, Malaysia. p. 2

  6. Ngobisa AIN, Ndongo PO, Doungous O, Ntsomboh-Ntsefong G, Njonje SW, Ehabe EE (2017) Characterization of Pestalotiopsis microspora, causal agent of leaf blight on rubber (Hevea Brasiliensis) in Cameroon. In Proceedings of International Rubber Conference, International Rubber Research and Development Board and Indonesian Rubber Research Institute, Jakarta, Indonesia, 18th -20th October 2017. p. 436–447

  7. Pornsuriya C, Chairin T, Thaochan N, Sunpapao A (2020) Identification and characterization of Neopestalotiopsis fungi associated with a novel leaf fall disease of rubber trees (Hevea brasiliensis) in Thailand. J Phytopathol 168:1–12

    Article  Google Scholar 

  8. Li B, Liu X, Jimiao C, Feng Y, Huang G (2020) First report on Neopestalotiopsis aotearoa of rubber tree in China. Plant Dis 105(4):1223

    Article  Google Scholar 

  9. Narayanan C, Reju MJ (2020) Variation in susceptibility to Phyllosticta capitalensis-associated leaf disease among inter-specific hybrids, half-sibs and high-yielding clones of Para rubber tree (Hevea). Plant Pathol Quarantine 10(1):p100–p110

    Article  Google Scholar 

  10. Aliya SSS, Nusaibah SA, Murnita MM, Yun WM, Yusop MR (2022) Colletotrichum siamense and Pestalotiopsis Jesteri as potential pathogens of new rubber leaf spot disease via detached leaf assay. J Rubber Res 25:195–212

    Article  Google Scholar 

  11. Aizat SN (2019) Pestalotiopsis leaf spot disease: identification of fungal pathogen and Koch’s postulates. First Quarterly Report in Integrated Pest and Disease Management Unit, Malaysian Rubber Board, Kuala Lumpur, Malaysia. p. 1–12

  12. Anon (2022) Report of the Special Meeting of the IRRDB Plant Protection Experts on Pestalotiopsis Leaf Disease. International Rubber Research and Development Board (IRRDB) Kuala Lumpur, Malaysia, 15th December 2022

  13. Mohamad Zamir HI (2023) Identification of possible pathogens of Pestalotiopsis leaf fall disease (PLFD), In Paperwork of Special Colloquium for Confirmation, Malaysian Rubber Board, Kuala Lumpur, Malaysia. pp. 139

  14. Murnita MM (2019) Pestalotiopsis leaf disease of Hevea brasiliensis: experience of Malaysia. In Proceedings of the First Meeting of Technical Committee on Plant Protection, International Rubber Research and Development Board, Kuala Lumpur, Malaysia, 5th November 2019

  15. Malaysian R, Board MRB (2021) Improvement to the Previous MRB Clone Recommendation 2019. In Malaysian Rubber Board Clone Recommendation 2021: MRB Clone Recommendation. Malaysian Rubber Board, Kuala Lumpur, Malaysia. p. 2

  16. Shuib SS, Déon M, Mahyuddin MM, Izhar A, Fumanal B, Sunderasan E, Pujade-Renaud V (2015) Cassiicolin genes among Corynespora Cassiicola isolates from rubber plantations in Malaysia. J Rubber Res 18(2):109–126

    Google Scholar 

  17. Sangu SS, Muid S (2016) Effects of inoculum concentrations of Colletotrichum gloeosporioides on disease development and severity on leaves of rubber tree (Hevea brasiliensis). Borneo J Resource Sci Technol 6(1):50–54

    Article  Google Scholar 

  18. Eskridge KM (1995) Statistical analysis of disease reaction data using nonparametric methods. HortScience 30(3):478–481

    Article  Google Scholar 

  19. Patil I (2021) Visualizations with statistical details: the ‘ggstatsplot’ approach. J Open-Source Softw 6:1–5 61, Article 3167

    Article  Google Scholar 

  20. R Core Team (2020) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available at: https://www.R-project.org/

  21. Jeffreys H (1961) The Pearson laws. Theory of probability, vol 3. Oxford University Press, New York, USA, p 74

    Google Scholar 

  22. De Silva DD, Crous PW, Ades PK, Hyde KD, Taylor PW (2017) Life styles of Colletotrichum species and implications for plant biosecurity. Fungal Biology Reviews 31:155–168

    Article  Google Scholar 

  23. Newton AC, Fitt BD, Atkins SD, Walters DR, Daniell TJ (2010) Pathogenesis, parasitism and mutualism in the trophic space of microbe–plant interactions. Trends in Microbiology, Vol. 18, No. 8. p. 365–373

  24. Salvatore MM, Andolfi A, Nicoletti R (2020) The thin line between pathogenicity and endophytism: the case of Lasiodiplodia theobromae. Agriculture 10(10):1–22Article 488

    Article  Google Scholar 

  25. Paolinelli-Alfonso M, Villalobos-Escobedo JM, Rolshausen P, Herrera-Estrella A, Galindo-Sánchez C, López-Hernández JF, Hernandez-Martinez R (2016) Global transcriptional analysis suggests Lasiodiplodia theobromae pathogenicity factors involved in modulation of grapevine defensive response. BMC Genomics, Vol. 17, Article 615. p. 1–20

  26. Hiruma K (2019) Plant interactions with parasitic and beneficial Colletotrichum fungi under changing environmental conditions. J Gen Plant Pathol 85:468–470

    Article  CAS  Google Scholar 

  27. O’Connell RJ, Thon MR, Hacquard S, Amyotte SG, Kleemann J, Torres MF, Vaillancourt LJ (2012) Lifestyle transitions in plant pathogenic Colletotrichum fungi deciphered by genome and transcriptome analyses. Nat Genet 44(9):1060–1065

    Article  PubMed  PubMed Central  Google Scholar 

  28. Watanabe K, Motohashi K, Ono Y (2010) Description of Pestalotiopsis pallidotheae: a new species from Japan. Mycoscience 51:182–188

    Article  Google Scholar 

  29. Maharachchikumbura SS, Guo LD, Cai L, Chukeatirote E, Wu WP, Sun X, Hyde KD (2012) A multi-locus backbone tree for Pestalotiopsis, with a polyphasic characterization of 14 new species. Fungal Divers 56:95–129

    Article  Google Scholar 

  30. De Silva NI, Maharachchikumbura SSN, Thambugala KM, Bhat DJ, Karunarathna SC, Tennakoon DS, Hyde KD (2021) Morpho-molecular taxonomic studies reveal a high number of endophytic fungi from Magnolia candolli and M. Garrettii in China and Thailand. Mycosphere 12(1):p163–p237 Article. 3

    Article  Google Scholar 

  31. De Errasti A, Novas MV, Carmarán CC (2014) Plant-fungal association in trees: insights into changes in ecological strategies of Peroneutypa scoparia (Diatrypaceae). In Flora: Morphology, Distribution, Functional Ecology of Plants (Ed. Heilmeier, H.), Elsevier, Freiberg, Germany, Vol. 209. No. 12, p. 704–710

  32. De Silva NI, Brooks S, Lumyong S, Hyde KD (2019) Use of endophytes as biocontrol agents. Fungal Biology Reviews 33:133–148

    Article  Google Scholar 

  33. Norphanphoun C, Hongsanan S, Gentekaki E, Chen YJ, Kuo CH, Hyde KD (2020) Differentiation of species complexes in Phyllosticta enables better species resolution. Mycosphere 11(1):2542–2628

    Article  Google Scholar 

  34. Moran Lemir AH, Stadnik MJ, Buchenauer H, Canton NV (2000) In vitro production of ascospores and pathogenicity of Guignardia citricarpa, causal agent of citrus black spot. Summa Phytopathologica 26(3):374–376

    Google Scholar 

  35. Fergus CL (1954) The effect of temperature and nutrients upon spore germination of the oak wilt fungus. Mycologia 46(4):435–441

    Article  Google Scholar 

  36. Dos Santos RF, Spósito MB, Ayres M, Sosnowski M (2018) In vitro production of conidia of Elsinoë Ampelina, the causal fungus of grapevine anthracnose. Eur J Plant Pathol 152(3):815–821

    Article  Google Scholar 

  37. Savatin DV, Gramegna G, Modesti V, Cervone F (2014) Wounding in the plant tissue: the defense of a dangerous passage. Front Plant Sci 5:1–11Article 470

    Article  Google Scholar 

  38. Divon HH, Fluhr R (2007) Nutrition acquisition strategies during fungal infection of plants. FEMS Microbiol Lett 266(1):65–74

    Article  CAS  PubMed  Google Scholar 

  39. Abdullah AS, Moffat CS, Lopez-Ruiz FJ, Gibberd MR, Hamblin J, Zerihun A (2017) Host–multi-pathogen warfare: pathogen interactions in co-infected plants. Front Plant Sci 8:1–12Article 1806

    Article  Google Scholar 

  40. Saucedo-Picazo LE, Hernández-Montiel LG, Flores-Estévez N, Gerez-Fernández P, Argüello-Ortiz AF, Noa-Carrazana JC (2022) Coinfection and in vitro interaction of Lasiodiplodia pseudotheobromae and Pestalotiopsis mangiferae associated with dieback in branches of mango (Mangifera indica) Manila variety, in Veracruz, Mexico. Mexican J Phytopathol 40(3):308–329

    Google Scholar 

  41. Fayyaz A, Bonello P, Tufail MR, Amrao L, Habib A, Gai Y, Sahi ST (2018) First report of citrus withertip (tip dieback), a disease complex caused by Colletotrichum siamense and Lasiodiplodia iraniensis, on Citrus reticulata cv. Kinnow in Punjab, Pakistan. In Plant Disease (Ed. Burrows, M. E.), The American Phytopathological Society, Minnesota, USA, Vol. 102, No. 12. p. 1–3

  42. Adeniyi DO, Orisajo SB, Fademi OA, Adenuga OO, Dongo LN (2011) Physiological studies of fungi complexes associated with cashew diseases. J Agricultural Biol Sci 6(4):34–38

    Google Scholar 

  43. Niroshini-Gunasinghe WKR, Karunaratne AM (2009) Interactions of Colletotrichum musae and Lasiodiplodia theobromae and their biocontrol by Pantoea agglomerans and Flavobacterium Sp. Expression Crown rot Embul Banana Biocontrol 54:587–596

    Google Scholar 

  44. Xu XM, Parry DW, Nicholson P, Thomsett MA, Simpson D, Edwards SG, Tatnell J (2005) Predominance and association of pathogenic fungi causing Fusarium ear blight in wheat in four European countries. Eur J Plant Pathol 112:143–154

    Article  Google Scholar 

  45. Gan P, Narusaka M, Kumakura N, Tsushima A, Takano Y, Narusaka Y, Shirasu K (2016) Genus-wide comparative genome analyses of Colletotrichum species reveal specific gene family losses and gains during adaptation to specific infection lifestyles. In Genome biology and evolution (Eds. Eyre-Walker, A. & Katz, L. A.). Oxford University Press, Oxford, UK, Vol. 8, No. 5. p. 1467–1481

  46. Buiate EA, Xavier KV, Moore N, Torres MF, Farman ML, Schardl CL, Vaillancourt LJ (2017) A comparative genomic analysis of putative pathogenicity genes in the host-specific sibling species Colletotrichum graminicola and Colletotrichum sublineola. BMC Genomics, Vol. 18, Article 67. p. 1–24

  47. Liang X, Wang B, Dong Q, Li L, Rollins JA, Zhang R, Sun G (2018) Pathogenic adaptations of Colletotrichum fungi revealed by genome wide gene family evolutionary analyses. PLoS ONE 13(4):1–25

    Article  Google Scholar 

  48. Gan P, Ikeda K, Irieda H, Narusaka M, O’Connell RJ, Narusaka Y, Shirasu K (2013) Comparative genomic and transcriptomic analyses reveal the hemibiotrophic stage shift of Colletotrichum fungi. New Phytol 197:1236–1249

    Article  CAS  PubMed  Google Scholar 

  49. Rao S, Nandineni MR (2017) Genome sequencing and comparative genomics reveal a repertoire of putative pathogenicity genes in chilli anthracnose fungus Colletotrichum truncatum. Plos One, Vol. 12, No. 8. p. 1–31

  50. Baroncelli R, Amby DB, Zapparata A, Sarrocco S, Vannacci G, Le Floch G, Thon MR (2016) Gene family expansions and contractions are associated with host range in plant pathogens of the genus Colletotrichum. BMC Genomics, Vol. 17, Article 555. p. 1–17

  51. Belisário R, Aucique-Pérez CE, Abreu LM, Salcedo SS, Oliveira WD, Furtado GQ (2020) Infection by Neopestalotiopsis spp. occurs on unwounded eucalyptus leaves and is favoured by long periods of leaf wetness. Plant Pathol 69:194–204

    Article  Google Scholar 

  52. Nagata T, Ando Y, Hirota A (1992) Phytotoxins from tea gray blight fungi, Pestalotiopsis longiseta and Pestalotiopsis theae. Bioscience, Biotechnology, And Biochemistry, Vol. 56, No. 5. p. 810–811

  53. Venkatasubbaiah P, Van Dyke CG (1991) Phytotoxins produced by Pestalotiopsis oenotherae, a pathogen of evening primrose. Phytochemistry 30:1471–1474No. 5

    Article  CAS  Google Scholar 

  54. Türkkan M, Andolfi A, Zonno MC, Erper I, Perrone C, Cimmino A, Evidente A (2011) Phytotoxins produced by Pestalotiopsis Guepinii, the causal agent of hazelnut twig blight. Phytopathologia Mediterranea 50:154–158

    Google Scholar 

  55. Feng L, Han J, Wang J, Zhang AX, Miao YY, Tan NH, Wang Z (2020) Pestalopyrones A–D, four tricyclic pyrone derivatives from the endophytic fungus Pestalotiopsis neglecta S3. Phytochemistry, Vol. 179, Article 112505. p. 1–7

  56. Spoel SH, Koornneef A, Claessens SM, Korzelius JP, Van Pelt JA, Mueller MJ (2003) NPR1 modulates cross-talk between salicylate-and jasmonate-dependent defense pathways through a novel function in the cytosol. Plant Cell 15:760–770

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Cooper A, Latunde-Dada A, Woods-Tör A, Lynn J, Lucas J, Crute I (2008) Basic compatibility of Albugo candida in Arabidopsis thaliana and Brassica juncea causes broad-spectrum suppression of innate immunity. In Molecular Plant-Microbe Interactions (Ed. Friesen, T. L.), The American Phytopathological Society, Vol. 21, No. 6. p. 745–756

  58. Notz R, Maurhofer M, Dubach H, Haas D, Défago G (2002) Fusaric acid-producing strains of Fusarium oxysporum alter 2, 4-diacetylphloroglucinol biosynthetic gene expression in Pseudomonas fluorescens CHA0 in vitro and in the rhizosphere of wheat. Applied And Environmental Microbiology, Vol. 68, No. 5. p. 2229–2235

  59. Saunders M, Kohn LM (2008) Host-synthesized secondary compounds influence the in vitro interactions between fungal endophytes of maize. Appl Environ Microbiol Vol 74(1):136–142

    Article  CAS  Google Scholar 

  60. Friesen TL, Stukenbrock EH, Liu Z, Meinhardt S, Ling H, Faris JD (2006) Emergence of a new disease as a result of interspecific virulence gene transfer. Nature Genetics, Vol. 38, No. 8. p. 953–956

  61. Al-Naimi FA, Garrett KA, Bockus WW (2005) Competition, facilitation, and niche differentiation in two foliar pathogens. Oecologia 143:449–457

    Article  CAS  PubMed  Google Scholar 

  62. Jiang J, Abbott KC, Baudena M, Eppinga MB, Umbanhowar JA, Bever JD (2020) Pathogens and mutualists as joint drivers of host species coexistence and turnover: implications for plant competition and succession. Am Nat 195(4):591–602

    Article  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the MRB for funding of this research and the permission to publish this paper. Special appreciation goes to the supporting staff of the Integrated Pest and Disease Management Unit for their assistance during the experiment and data collection.

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The research was fully supported by the Malaysian Rubber Board.

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  1. Malaysian Rubber Board, RRIM Research Station Sungai Buloh, Sungai Buloh, Selangor, 47000, Malaysia

    Mohamad Zamir Hadi Ismail, Murnita Mohmad Mahyudin, Aizat Shamin Noran, Adam Malik Ahmad Zambri, Nor Afiqah Maiden, Safiah Atan & Mohd Nasaruddin Mohd Aris

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  1. Mohamad Zamir Hadi Ismail
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Hadi Ismail, M.Z., Mahyudin, M.M., Noran, A.S. et al. Identification and characterisation of causal pathogens of Pestalotiopsis leaf fall disease in Hevea brasiliensis using a detached leaf technique. J Rubber Res 27, 159–173 (2024). https://doi.org/10.1007/s42464-024-00243-9

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