Mangrove Dieback and Leaf Disease in Sonneratia apetala and Sonneratia caseolaris in Vietnam

Even though survival rates for mangrove restoration in Vietnam have often been low, there is no information on fungal pathogens associated with mangrove decline in Vietnam. Therefore, this research was undertaken to assess the overall health of mangrove afforestation in Thanh Hoa Province and fungal pathogens associated with tree decline. From a survey of 4800 Sonneratia trees, the incidence of disorders was in the order of pink leaf spot > shoot dieback > black leaf spot for S. caseolaris and black leaf spot > shoot dieback > pink leaf spot for S. apetala. Approximately 12% of S. caseolaris trees had both pink leaf spot and shoot dieback, while only 2% of S. apetala trees had black leaf spot and shoot dieback. Stem and leaf samples were taken from symptomatic trees and fungi were cultured in vitro. From ITS4 and ITS5 analysis, four main fungal genera causing leaf spots and shoot dieback on the two Sonneratia species were identified. The most frequently isolated fungal taxa were Curvularia aff. tsudae (from black leaf spot), Neopestalotiopsis sp.1 (from stem dieback), Pestalotiopsis sp.1 (from pink leaf spot), and Pestalotiopsis sp.4a (from black leaf spot). The pathogenicity of the four isolates was assessed by under-bark inoculation of S. apetala and S. caseolaris seedlings in a nursery in Thai Binh Province. All isolates caused stem lesions, and Neopestalotiopsis sp.1 was the most pathogenic. Thus, investigation of fungal pathogens and their impact on mangrove health should be extended to other afforestation projects in the region, and options for disease management need to be developed for mangrove nurseries.

1. Introduction

2. Materials and Methods

2.1. Study Area and Canopy Assessment

2.2. Sample Collection

2.3. Fungal Isolation

2.4. Fungal Identification

2.5. Pathogenicity Tests

2.6. Statistical Analysis

3. Results

3.1. Tree Health Survey

3.2. Symptoms of Disease on Sonneratia Species

3.3. Taxonomy

3.4. Morphology of the Main Taxa

3.5. Pathogenicity Trial

4. Discussion

5. Conclusions

References

1. Sherman, R.E.; Fahey, T.J.; Martinez, P. Hurricane Impacts on a Mangrove Forest in the Dominican Republic: Damage Patterns and Early Recovery 1. Biotropica 2001, 33, 393–408. [CrossRef]

2. Kathiresan, K.; Bingham, B.L. Biology of Mangroves and Mangrove Ecosystems. Adv. Mar. Biol. 2001, 40, 81–251.

3. Taylor, M.; Ravilious, C.; Green, E.P. Mangroves of East Africa; UNEP–WCMC: Cambridge, UK, 2003.

4. Martinuzzi, S.; Gould, W.A.; Lugo, A.E.; Medina, E. Conversion and Recovery of Puerto Rican Mangroves: 200 years of change. For. Ecol. Manag. 2009, 257, 75–84. [CrossRef]

5. Abuodha, P.; Kairo, J. Human-induced Stresses on Mangrove Swamps along the Kenyan Coast. Hydrobiologia 2001, 458, 255–265. [CrossRef]

6. Ellison, A.M. Mangrove Communities. In Marine Community Ecology; Sinauer Associates, Inc.: Sunderland, MA, USA, 2001; pp. 423–442.

7. Spalding, M. World Atlas of Mangroves; Routledge: London, UK, 2010.

8. Smith, T.J., III; Robblee, M.B.; Wanless, H.R.; Doyle, T.W. Mangroves, Hurricanes, and Lightning Strikes: Assessment of Hurricane Andrew Suggests an Interaction across Two Differing Scales of Disturbance. BioScience 1994, 44, 256–262. [CrossRef]

9. Norm, D.; Jock, M.; John, K.; Grant, S.; Rob, C.; Apanie, W.; Ysabel, C. Assessing the Gulf of Carpentaria Mangrove Dieback 2017–2019; James Cook University: Douglas, Australia, 2020; p. 249.

10. Van, O.P.; Duke, N. Extreme Weather Likely behind Worst Recorded Mangrove Dieback in Northern Australia. Conversation 2017, 14, 1–6.

11. Gilbert, G.S. Evolutionary Ecology of Plant Diseases in Natural Ecosystems. Annu. Rev. Phytopathol. 2002, 40, 13–43. [CrossRef]

12. Hulcr, J.; Dunn, R.R. The Sudden Emergence of Pathogenicity in Insect–fungus Symbioses Threatens Naive Forest Ecosystems. Proc. R. Soc. B Biol. Sci. 2011, 278, 2866–2873. [CrossRef]

13. Hyde, K.; Cannon, P. Polystigma sonneratiae Causing Leaf Spots on The Mangrove Genus Sonneratia. Aust. Syst. Bot. 1992, 5, 415–420. [CrossRef]

14. Jones, E.G.; Pang, K.-L. Tropical Aquatic Fungi. Biodivers. Conserv. 2012, 21, 2403–2423. [CrossRef]

15. Jones, E.G.; Pang, K.-L.; Abdel-Wahab, M.A.; Scholz, B.; Hyde, K.D.; Boekhout, T.; Ebel, R.; Rateb, M.E.; Henderson, L.; Sakayaroj, J. An Online Resource for Marine Fungi. Fungal Divers. 2019, 96, 347–433. [CrossRef]

16. Alias, S.; Jones, E. Colonization of Mangrove Wood by Marine Fungi at Kuala Selangor Mangrove Stand, Malaysia. Fungal Divers. 2000, 5, 9–21.

17. Kohlmeyer, J.; Kohlmeyer, E. Marine Fungi From Tropical America and Africa. Mycologia 1971, 63, 831–861. [CrossRef] [PubMed]

18. Jones, E.G.; Abdel-Wahab, M.A. Marine Fungi from the Bahamas Islands. Bot. Mar. 2005, 48, 356–364.

19. Sarma, V.; Hyde, K.D. A review on Frequently Occurring Fungi in Mangroves. Fungal Divers 2001, 8, 1–34.

20. Steinke, T. Mangrove Fungi on Dead Proproots of Rhizophora mucronata at Three Localites in South Africa. S. Afr. J. Bot. 2000, 66, 91–95. [CrossRef]

21. Osorio, J.A.; Wingfield, M.J.; Roux, J. A Review of Factors Associated with Decline and Death of Mangroves, with Particular Reference to Fungal Pathogens. S. Afr. J. Bot. 2016, 103, 295–301. [CrossRef]

22. Xie, S.; He, H.; Yang, R.; Xu, Z.; He, J.; Lu, H. First Report of Leaf Spot Caused by Corynespora cassiicola on Acanthus ilicifolius in China. Plant Dis. 2021, 105, 509. [CrossRef]

23. Stevens, F.L. New or Noteworthy Porto Rican Fungi. Bot. Gaz. 1920, 70, 399–402. [CrossRef]

24. Rahman, M.A. Top Dying of Sundri (Heritiera fomes) and Its Impact on the Regeneration and Management in the Mangrove Forests of Sunderbans in Bangladesh. In Proceedings of the Impact of Diseases and Insect Pests in Tropical Forests, Peechi City, India, 23–26 November 1993; pp. 117–133.

25. Yan, T.L.; Zhu, X.J.; Yang, Z.D.; Long, Q.H.; Yang, H.; Chang, S.Q. New Records of Celoporthe guangdongensis and Cytospora rhizophorae on Mangrove Apple in China. Biodivers. Data J. 2020, 8, 15.

26. McMillan, J.R. Studies of a Recently Described Cercospora on Rhizophora mangle. Plant Dis. Report. 1964, 48, 909–991.

27. Creager, D.B. A New Cercospora on Rhizophora mangle. Mycologia 1962, 54, 536–539. [CrossRef]

28. Osorio, J.A.; Crous, C.J.; Wingfield, M.J.; De Beer, Z.W.; Roux, J. An assessment of Mangrove Diseases and Pests in South Africa. For. Int. J. For. Res. 2017, 90, 343–358. [CrossRef]

29. Osorio, J.A.; Crous, C.J.; De Beer, Z.W.; Wingfield, M.J.; Roux, J. Endophytic Botryosphaeriaceae, Including Five New Species, Associated with Mangrove Trees in South Africa. Fungal Biol. 2017, 121, 361–393. [CrossRef] [PubMed]

30. Wier, A.M.; Tattar, T.A.; Klekowski, E.J., Jr. Disease of Red Mangrove (Rhizophora mangley) in Southwest Puerto Rico Caused by Cytospora rhizophorae 1. Biotropica 2000, 32, 299–306.

31. Kohlmeyer, J. Ecological Notes on Fungi in Mangrove Forests. Trans. Br. Mycol. Soc. 1969, 53, 237–250. [CrossRef]

32. Teas, H.; McEwan, R. An Epidemic Dieback Gall Disease of Rhizophora mangroves in The Gambia, West Africa. Plant Dis. 1982, 66, 522–523. [CrossRef]

33. Sakayaroj, J.; Preedanon, S.; Suetrong, S.; Klaysuban, A.; Jones, E.G.; Hattori, T. Molecular Characterization of Basidiomycetes Associated with The Decayed Mangrove Tree Xylocarpus granatum in Thailand. Fungal Divers. 2012, 56, 145–156. [CrossRef]

34. Pegg, K.; Gillespie, N.; Forsberg, L. Phytophthora sp. Associated with Mangrove Death in Central Coastal Queensland. Australas. Plant Pathol. 1980, 9, 6–7. [CrossRef]

35. Xu, J.; Kjer, J.; Sendker, J.; Wray, V.; Guan, H.; Edrada, R.; Lin, W.; Wu, J.; Proksch, P. Chromones From the Endophytic Fungus Pestalotiopsis sp. Isolated from the Chinese Mangrove Plant Rhizophora mucronata. J. Nat. Prod. 2009, 72, 662–665. [CrossRef]

36. Olexa, M.; Freeman, T. Occurrence of Three Unrecorded Diseases on Mangroves in Florida. In Proceedings of the International Symposium on Biology and Management of Mangroves, Honolulu, HI, USA, 8–11 October 1974; pp. 688–692.

37. Shivas, R.; Young, A.; Crous, P. Pseudocercospora avicenniae RG Shivas, AJ Young & Crous, sp. nov. Persoonia 2009, 23, 193.

38. Rafael, A.; Calumpong, H.P. Fungal Infections of Mangroves in Natural Forests and Reforestation Sites from Philippines. Aquac. Aquar. Conserv. Legis. 2019, 12, 2062–2074.

39. Hong, P.N.; San, H.T. Mangroves of Vietnam; IUCN: Bangkok, Thailand, 1993; pp. 150–180.

40. Ministry of Agriculture and Rural Development. Announcement of National Forest Status in 2019; MARD: Hanoi, Vietnam, 2020.

41. Hai, N.; Dell, B.; Phuong, V.; Harper, R. Towards a More Robust Approach for The Restoration of Mangroves in Vietnam. Ann. For. Sci. 2020, 77, 1–18. [CrossRef]

42. Nguyen, H.H.; Nghia, N.H.; Nguyen, H.T.T.; Le, A.T.; Tran, L.T.N.; Duong, L.V.K.; Bohm, S.; Furniss, M.J. Classification Methods for Mapping Mangrove Extents and Drivers of Change in Thanh Hoa Province, Vietnam During 2005–2018. For. Soc. 2020, 4,225–242. [CrossRef]

43. Vietnam Admistration of Forestry. Protective Forest in Coastal Is Unexplained Death. Available online: http://bentre.tintuc.vn/tin-tuc/ben-tre-rung-phong-ho-dac-dung-ven-bien-chet-chua-ro-nguyen-nhan.html (accessed on 1 July 2021).

44. Baotainguyenvamoitruong. Proposing Scientific Research to Find the Cause of Death of Mangroves in Ben Tre. Available online: https://baotainguyenmoitruong.vn/moi-truong/ben-tre-de-xuat-nghien-cuu-khoa-hoc-tim-nguyen-nhan-rung-chet-1258495.html (accessed on 1 July 2021).

45. Sggp.org.vn. Death of Rhizophora in Mass: Identify reasons (Vietnamese). Available online: http://www.sggp.org.vn/vu-cayduoc-chet-hang-loat-xac-dinh-loai-sinh-vat-la-gay-hai-451729.html (accessed on 1 July 2021).

46. Antv.gov.vn. Death of Mangroves in Mass in Ha Tinh (Vietnamese). Available online: http://www.antv.gov.vn/tin-tuc/xa-hoi/rung-cay-ngap-man-o-ha-tinh-chet-hang-loat-186342.html (accessed on 1 July 2021)

47. Nguyen, H.T.T.; Hardy, G.E.; Le, T.V.; Nguyen, H.Q.; Nguyen, H.H.; Nguyen, T.V.; Dell, B. Mangrove Forest Landcover Changes in Coastal Vietnam: A Case Study from 1973 to 2020 in Thanh Hoa and Nghe An Provinces. Forests 2021, 12, 637. [CrossRef]

48. Doyle, J.J.; Doyle, J.L. A Rapid DNA Isolation Procedure for Small Quantities of Fresh Leaf Tissue. Phytochem. Bull. 1987, 19,11–15.

49. Khan, A.M.; Bhadauria, S. Molecular Characterization of Keratin Degrading Fungi Isolated from Semi-arid Soil by PCR Using ITS4 and ITS5 Primers. J. King Saud Univ.-Sci. 2019, 31, 1418–1423. [CrossRef]

50. White, T.J.; Bruns, T.; Lee, S.; Taylor, J. Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. In PCR Protocols: A Guide to Methods and Applications; Academic Press: New York, NY, USA, 1990; Volume 18, pp. 315–322.

51. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular Evolutionary Genetics Analysis Across Computing Platforms. Mol. Biol. Evol. 2018, 35, 1547–1549. [CrossRef]

52. Allen, P.; Bennett, K.; Heritage, B. SPSS Statistics Version 22: A Practical Guide; Cengage Learning Australia Pty Limited: Sydney, Australia, 2014; pp. 200–250.

53. Shamsi, S.; Hosen, S.; Ahmed, A. Fungi Associated with Leaves of Sonneratia apetala Buch. Ham and Sonneratia caseolaris (L.) Engler from Rangabali Coastal Zone of Bangladesh. Dhaka Univ. J. Biol. Sci. 2018, 27, 155–162. [CrossRef]

54. Abbas, M.F.; Rafiq, M.; Al-Sadi, A.M.; Alfarraj, S.; Alharbi, S.A.; Arif, M.; Ansari, M.J. Molecular Characterization of Leaf Spot caused by Alternaria alternata on Buttonwood (Conocarpus erectus L.) and Determination of Pathogenicity by a Novel Disease Rating Scale. PLoS ONE 2021, 16, e0251471. [CrossRef]

55. Tattar, T.; Klekowski, E.; Stern, A. Dieback and Mortality in Red Mangrove, Rhizophora mangle L., in Southwest Puerto Rico. Arboric. J. 1994, 18, 419–429. [CrossRef]

56. Liu, A.; Wu, X.; Xu, T. Research advances in endophytic fungi of mangrove. J. Appl. Ecol. 2007, 18, 912–918.

57. Suryanarayanan, T.; Kumaresan, V. Endophytic fungi of some halophytes from an estuarine mangrove forest. Mycol. Res. 2000, 104, 1465–1467. [CrossRef]

58. Chaeprasert, S.; Piapukiew, J.; Whalley, A.J.; Sihanonth, P. Endophytic Fungi from Mangrove Plant Species of Thailand: Their Antimicrobial and Anticancer Potentials. Bot. Mar. 2010, 53, 555–564. [CrossRef]

59. Xing, X.; Guo, S. Fungal Endophyte Communities in four Rhizophoraceae Mangrove Species on The South Coast of China. Ecol. Res. 2011, 26, 403–409. [CrossRef]

60. Khan, A.A.H. Endophytic Fungi and Their Impact on Agroecosystems. In Medicinal Plants: Biodiversity, Sustainable Utilization and Conservation; Springer: Singapore, 2020; pp. 443–499.

61. Nurunnabi, T.R.; Sabrin, F.; Sharif, D.I.; Nahar, L.; Sohrab, M.H.; Sarker, S.D.; Rahman, S.M.; Billah, M.M. Antimicrobial Activity of Endophytic Fungi Isolated From the Mangrove Plant Sonneratia apetala (Buch.-Ham) from the Sundarbans Mangrove Forest. Adv. Tradit. Med. 2020, 40, 1–7. [CrossRef]

62. Goudarzi, A.; Moslehi, M. Distribution of a Devastating Fungal Pathogen in Mangrove Forests of Southern Iran. Crop Prot. 2020, 128, 104987. [CrossRef]

63. Debbab, A.; Aly, A.H.; Proksch, P. Mangrove Derived Fungal Endophytes–a Chemical and Biological Perception. Fungal Divers. 2013, 61, 1–27. [CrossRef]

64. Ebrahim, W.; Aly, A.H.; Mándi, A.; Totzke, F.; Kubbutat, M.H.; Wray, V.; Lin, W.H.; Dai, H.; Proksch, P.; Kurtán, T. Decalactone Derivatives from Corynespora cassiicola, an Endophytic Fungus of the Mangrove Plant Laguncularia racemosa. Eur. J. Org. Chem. 2012, 2012, 3476–3484. [CrossRef]

65. Cheng, Z.S.; Tang, W.C.; Xu, S.L.; Sun, S.F.; Huang, B.Y.; Yan, X.; Chen, Q.J.; Lin, Y.C. First Report of An endophyte (Diaporthe phaseolorum var. sojae) from Kandelia candel. J. For. Res. 2008, 19, 277–282. [CrossRef]

66. Xing, X.; Chen, J.; Xu, M.; Lin, W.; Guo, S. Fungal Endophytes Associated with Sonneratia (Sonneratiaceae) Mangrove Plants on the South Coast of China. For. Pathol. 2011, 41, 334–340. [CrossRef]

67. Olexa, M.; Freeman, T. A Gall Disease of Red Mangrove Caused by Cylindrocarpon didymum. Plant Dis. Report. 1978, 62, 283–286.

68. Chi, W.-C.; Chen, W.; He, C.-C.; Guo, S.-Y.; Cha, H.-J.; Tsang, L.M.; Ho, T.W.; Pang, K.-L. A Highly Diverse Fungal Community Associated with Leaves of the Mangrove Plant Acanthus ilicifolius var. xiamenensis Revealed by Isolation and Metabarcoding Analyses. PeerJ 2019, 7, e7293. [CrossRef]

69. Cui, H.; Lin, Y.; Luo, M.; Lu, Y.; Huang, X.; She, Z. Diaporisoindoles A–C: Three isoprenylisoindole alkaloid derivatives from the mangrove endophytic fungus Diaporthe sp. SYSU-HQ3. Org. Lett. 2017, 19, 5621–5624. [CrossRef] [PubMed]

70. Norphanphoun, C.; Jayawardena, R.; Chen, Y.; Wen, T.; Meepol, W.; Hyde, K. Morphological and Phylogenetic Characterization of Novel Pestalotioid Species Associated with Mangroves in Thailand. Mycosphere 2019, 10, 531–578. [CrossRef]

71. Hamzah, T.N.T.; Lee, S.Y.; Hidayat, A.; Terhem, R.; Faridah-Hanum, I.; Mohamed, R. Diversity and Characterization of Endophytic Fungi Isolated from the Tropical Mangrove Species, Rhizophora mucronata, and Identification of Potential Antagonists Against the Soil-Borne Fungus, Fusarium solani. Front. Microbiol. 2018, 9, 1707. [CrossRef] [PubMed]

72. Zhou, J.; Diao, X.; Wang, T.; Chen, G.; Lin, Q.; Yang, X.; Xu, J. Phylogenetic Diversity and Antioxidant Activities of Culturable Fungal Endophytes Associated with the Mangrove Species Rhizophora stylosa and R. mucronata in the South China Sea. PLoS ONE 2018, 13, e0197359. [CrossRef] [PubMed]

73. Deshmukh, S.K.; Gupta, M.K.; Prakash, V.; Reddy, M.S. Mangrove-Associated Fungi: A Novel Source of Potential Anticancer Compounds. J. Fungi 2018, 4, 101. [CrossRef]

74. Ha, N.T.; Ngat, H.T.; Ha, N.T.T.; Hoi, P.X. Study on Pathogenicity of Colletotrichum gloeosporioides on Coffee in North of Vietnam. Biology 2011, 33, 67–73. [CrossRef]

75. Tran, N.B.B. Isolation and Identification of Fungal Diseases in Rice (Oryza sativa) from Mekong Delta. Ph.D. Thesis, International University-HCMC, Ho Chi Minh City, Vietnam, 2019.

76. Ngoc, N.; Phong Nguyen, N.; An, P.; Woolf, A.; Fullerton, R. Effect of Storage Temperatures on Postharvest Diseases of Dragon Fruits (Hylocereus undatus Haw.) in the Mekong Delta Region, Vietnam. In Proceedings of the III Asia Pacific Symposium on Postharvest Research, Education and Extension: APS2014 1213, Hochiminh City, Vietnam, 8 December 2014; pp. 453–460.

77. Yen, T.T.N.; Thanh, N.V.; Phong, N.V. Research on the Causative Agent of Rambutan fruit rot (Nephelium lappaceum L.) after Harvest in the Mekong Delta. J. Sci. Technol. 2017, 13, 1868–1873.

78. Dung, L.; Dienb, N.T.; Daic, P.H.; Tuana, P.N. The First Report of Pestalofiopsis sp. Causing Crown Rot Disease on Strawberries in Dalat. Dalat Univ. J. Sci. 2016, 6, 364–376. [CrossRef]

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► See detail:  Mangrove Dieback and Leaf Disease in Sonneratia apetala and Sonneratia caseolaris in Vietnam

Huong Thi Thuy Nguyen 1,2, Giles E. St. J. Hardy 1 , Tuat Van Le 3, Huy Quoc Nguyen 3, Duc Hoang Le 4 ,
Thinh Van Nguyen 2 and Bernard Dell 1,5,*
1 Agriculture and Forest Sciences, Murdoch University, Murdoch 6150, Australia;
huong.nguyen@murdoch.edu.au (H.T.T.N.); G.Hardy@murdoch.edu.au (G.E.S.J.H.)
2 Silviculture Research Institute, Vietnamese Academy of Forest Sciences, Duc Thang, Bac Tu Liem,
Ha Noi 11910, Vietnam; nguyenthinhfsiv@gmail.com
3 Institute of Ecology and Works Protection, Vietnam Academy for Water Resources, 267 Chua Boc, Dong Da,
Ha Noi 11515, Vietnam; tuatwip@gmail.com (T.V.L.); huy_ctcr@yahoo.com (H.Q.N.)
4 Institute of Biotechnology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Nghia Do,
Cau Giay, Ha Noi 11350, Vietnam; lh.duc@ibt.ac.vn
5 Forest Protection Research Centre, Vietnamese Academy of Forest Sciences, Duc Thang, Bac Tu Liem,
Ha Noi 11910, Vietnam
* Correspondence: B.Dell@murdoch.edu.au; Tel.: +61-427-788-820

Forests 2021, 12, 1273. https://doi.org/10.3390/f12091273                                                             https://www.mdpi.com/journal/forests