Skip to main navigation menu Skip to main content Skip to site footer
Plant Science Today (PST)

Research Articles

Vol. 12 No. 1 (2025)

Molecular identification and characterization of pathogenicity of fungal pathogens associated with crown rot disease in banana from Odisha, India

DOI
https://doi.org/10.14719/pst.4151
Submitted
21 June 2024
Published
17-01-2025 — Updated on 25-01-2025
Versions

Abstract

Crown rot disease on banana varieties (Champa, Patakpura and G9) was identified in Odisha's 10 coastal districts during the post-harvest storage stage in 2022-23. Five crown fragments were taken for isolation from each variety of bananas based on symptom developments. Agar plate and standard blotter technique isolation procedures have been used to isolate 1210 fungal colonies, of which 584 representative colonies were purified from banana crown fragments. Different fungal colonies were purified, characterized and identified using morphological and molecular methods from G9 (244), Patakpura (205) and Champa (135) fungal colonies. Morphological identification of Fusarium and Colletotrichum isolates was madeusing colony morphology, pigmentation, the growth rate on the PDA plate and microscopic observation. Molecular identification of Fusarium and Colletotrichum isolates was done by amplifying the internal transcribed spacer (ITS) region of the conserved ribosomal DNA, which includes 5.8S rDNA. Fusarium verticillioides, Fusarium equiseti, Fusarium oxysporum and Colletotrichum gloeosporioides were the 4 fungal isolates identified. The sequences of these fungi were deposited in the GenBank database (NCBI) under the accession numbers OQ363325 (Fusarium verticillioides), OP735534 (Fusarium equiseti), OQ438654 (Fusarium oxysporum) and PP448030 (Colletotrichum gloeosporioides). The neighbor-joining method was used to conduct phylogenetic analysis on the gene sequences. Pathogenicity test results revealed that all 4 isolated fungal strains produced similar crown rot symptoms on 3 varieties of banana fruits. Disease severity index and AUDPC indicated that the Fusarium strain is the most aggressive crown rot pathogen and the G9 is the more susceptible banana variety to crown rot pathogens in Odisha.

References

  1. Yadav SK, Babu S, Yadav MK, Singh K, Yadav GS, Pal S. A review of organic farming for sustainable agriculture in Northern India. International Journal of Agronomy. 2013;2013(1):718145. https://doi.org/10.1155/2013/718145
  2. Alvindia DG. Improving control of crown rot disease and quality of pesticide-free banana fruit by combining Bacillus amylolique-faciens DGA14 and hot water treatment. European Journal of Plant Pathology. 2013;136(1):183-91. https://doi.org/10.1007/s10658-012-0154-7
  3. Lassois L, Jijakli MH, Chillet M, De Lapeyre de Bellaire L. Crown rot of bananas: preharvest factors involved in postharvest dis-ease development and integrated control methods. Plant Dis-ease. 2010 ;94(6):648-58. https://doi.org/10.1094/PDIS-94-6-0648
  4. Alvindia DG, Kobayashi T, Yaguchi Y, Natsuaki KT. Symptoms and the associated fungi of postharvest diseases on non-chemical bananas imported from the Philippines. Japanese Journal of Tropical Agriculture. 2000;44(2):87-93. https://doi.org/10.11248/jsta1957.44.87
  5. Alvindia DG, Kobayashi T, Tanda S. Identification of fungi isolat-ed from non-chemical banana fruits and farms in the Philip-pines. Journal of Agricultural Science-Tokyo-University of Agri-culture. 2002;47(2):78-97.
  6. Ewané CA, Lassois L, Brostaux Y, Lepoivre P, De Lapeyre de Bel-laire L. The susceptibility of bananas to crown rot disease is influenced by geographical and seasonal effects. Canadian Jour-nal of Plant Pathology. 2013;35(1):27-36. http://dx.doi.org/10.1080/07060661.2012.733731
  7. Triest D, Hendrickx M. Postharvest disease of banana caused by Fusarium musae: a public health concern? PLoS Pathogens. 2016;12(11):e1005940. https://doi.org/10.1371/journal.ppat.1005940
  8. Deltour P, França SC, Pereira OL, Cardoso I, De Neve S, Debode J, Höfte M. Disease suppressiveness to Fusarium wilt of banana in an agroforestry system: influence of soil characteristics and plant community. Agriculture, Ecosystems and Environment. 2017; 239:173-81. https://doi.org/10.1016/j.agee.2017.01.018
  9. Kamel MA, Cortesi P, Saracchi M. Etiological agents of crown rot of organic bananas in Dominican Republic. Postharvest Biology and Technology. 2016;120:112-20. https://doi.org/10.1016/j.postharvbio.2016.06.002
  10. Moretti A, Mulè G, Susca A, González-Jaén MT, Logrieco A. Toxin profile, fertility and AFLP analysis of Fusarium verticillioides from banana fruits. Molecular Diversity and PCR-detection of Toxigen-ic Fusarium Species and Ochratoxigenic Fungi: Under the aegis of COST Action 835 ‘Agriculturally Important Toxigenic Fungi 1998–2003’, EU project (QLK1-CT-1998-01380) and the ISPP ‘Fusarium Committee’. 2004:601-09. https://doi.org/10.1007/978-1-4020-2285-2_14
  11. Leslie JF, Summerell BA. The Fusarium laboratory manual. John Wiley and Sons; 2008.
  12. Singha IM, Kakoty Y, Unni BG, Das J, Kalita MC. Identification and characterization of Fusarium sp. using ITS and RAPD causing Fusarium wilt of tomato isolated from Assam, North East India. Journal of Genetic Engineering and Biotechnology. 2016;14(1):99-105. https://doi.org/10.1016/j.jgeb.2016.07.001
  13. Menezes JP, Lupatini M, Antoniolli ZI, Blume E, Junges E, Manzoni CG. Genetic variability in rDNA ITS region of Trichoder-ma spp. (biocontrol agent) and Fusarium oxysporum f. sp. chry-santhemi isolates. Ciencia e Agrotecnologia. 2010;34:132-39. https://doi.org/10.1590/S1413-70542010000100017
  14. Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research. 1990;18(22):6531-35. https://doi.org/10.1093/nar/18.22.6531
  15. Bayraktar HA, Dolar FS, Maden S. Use of RAPD and ISSR markers in detection of genetic variation and population structure among Fusarium oxysporum f. sp. ciceris isolates on chickpea in Turkey. Journal of Phytopathology. 2008;156(3):146-54. https://doi.org/10.1111/j.1439-0434.2007.01319.x
  16. Guleria S, Aggarwal R, Thind TS, Sharma TR. Morphological and pathological variability in rice isolates of Rhizoctonia solani and molecular analysis of their genetic variability. Journal of Phyto-pathology. 2007;155(11-12):654-61. https://doi.org/10.1111/j.1439-0434.2007.01291.x
  17. Lievens B, Claes L, Vakalounakis DJ, Vanachter AC, Thomma BP. A robust identification and detection assay to discriminate the cucumber pathogens Fusarium oxysporum f. sp. cucumerinum and f. sp. radicis-cucumerinum. Environmental Microbiology. 2007;9(9):2145-61. https://doi.org/10.1111/j.1462-2920.2007.01329.x
  18. AlvesAlves-Santos FM, Ramos B, García García-Sánchez MA, Eslava AP, Díaz Díaz-Mínguez JM. A DNA-based procedure for in planta detection of Fusarium oxysporum f. sp. phaseoli. Phytopathology. 2002;92(3):237-44. https://doi.org/10.1094/PHYTO.2002.92.3.237
  19. Sonker N, Pandey AK, Singh P. Efficiency of Artemisia nilagirica (Clarke) Pamp. essential oil as a mycotoxicant against posthar-vest mycobiota of table grapes. Journal of the Science of Food and Agriculture. 2015;95(9):1932-39. https://doi.org/10.1002/jsfa.6901
  20. Cai L, Hyde KD, Taylor PW, Weir B, Waller J, Abang MM, et al. A polyphasic approach for studying Colletotrichum. Fungal Diversi-ty. 2009;39(1):183-204.
  21. Cenis JL. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Research. 1992;20(9):2380. https://doi.org/10.1093%2Fnar%2F20.9.2380
  22. White TJ, Bruns T, Lee SJ, Taylor J. Amplification and direct se-quencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: A Guide to Methods and Applications. 1990;18(1):315-22. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
  23. Tamura K, Stecher G, Kumar S. MEGA11: molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution. 2021;38(7):3022-27. https://doi.org/10.1093/molbev/msab120
  24. Baró Robaina Y, González Marrero I, Lorenzo Nicao ME, Castañeda Ruiz RF, Li DW, Ponce de la Cal A, et al. First descrip-tion of Simplicillium lanosoniveum, a potential antagonist of the coffee leaf rust from Cuba. Applied Microbiology. 2024;4(1):275-83. https://doi.org/10.3390/applmicrobiol4010018
  25. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39(4):783-91. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x
  26. AbdAbd-Alla MA, El El-Gamal NG, El El-Mougy NS, Abdel Abdel-Kader MM. Post-harvest treatments for controlling crown rot disease of Williams banana fruits (Musa acuminata L.) in Egypt. Plant Pathology and Quarantine. 2014;4(1):1-2. https://doi.org/10.5943/ppq/4/1/1
  27. Khuna S, Kumla J, Thitla T, Nuangmek W, Lumyong S, Suwanna-rach N. Morphology, molecular identification and pathogenicity of two novel Fusarium species associated with postharvest fruit rot of cucurbits in Northern Thailand. Journal of Fungi. 2022;8(11):1135. https://doi.org/10.3390/jof8111135
  28. Alvindia DG, Kobayashi T, Natsuaki KT, Tanda S. Inhibitory influ-ence of inorganic salts on banana postharvest pathogens and preliminary application to control crown rot. Journal of General Plant Pathology. 2004;70:61-65. https://doi.org/10.1007/s10327-003-0084-5
  29. Campbell CL, Madden LV. Introduction to plant disease epidemi-ology. 1990.
  30. de Capdeville G, Wilson CL, Beer SV, Aist JR. Alternative disease control agents induce resistance to blue mould in harvested ‘Red Delicious’ apple fruit. Phytopathology. 2002;92(8):900-08. https://doi.org/10.1094/PHYTO.2002.92.8.900
  31. Lukezic FL, Kaiser WJ, Martinez MM. The incidence of crown rot of boxed bananas in relation to microbial populations of the crown tissue. Canadian Journal of Botany. 1967;45(4):413-21. https://doi.org/10.1139/b67-041
  32. Silva EG, Silva JA, Barros AP, Santos AK, Michereff SJ, Correia KC. Fusarium kalimantanense causing crown rot on banana in Brazil. Journal of Plant Pathology. 2023;105(3):1199. https://doi.org/10.1007/s42161-023-01401-5
  33. Rabha AJ, Naglot A, Sharma GD, Gogoi HK, Gupta VK, Shreemali DD, Veer V. Morphological and molecular diversity of endophytic Colletotrichum gloeosporioides from tea plant, Camellia sinensis (L.) O. Kuntze of Assam, India. Journal of Genetic Engineering and Biotechnology. 2016;14(1):181-87. https://doi.org/10.1016/j.jgeb.2015.12.003
  34. Zainudin NA, Hamzah FA, Kusai NA, Zambri NS, Salleh S. Charac-terization and pathogenicity of Fusarium proliferatum and Fusarium verticillioides, causal agents of Fusarium ear rot of corn. Turkish Journal of Biology. 2017;41(1):220-30. https://doi.org/10.3906/biy-1606-25
  35. Hami A, Rasool RS, Khan NA, Mansoor S, Mir MA, Ahmed N, Masoodi KZ. Morpho-molecular identification and first report of Fusarium equiseti in causing chilli wilt from Kashmir (Northern Himalayas). Scientific Reports. 2021;11(1):3610. https://doi.org/10.1038/s41598-021-82854-5
  36. AguilarAguilar-Hawod KG, de la Cueva FM, Cumagun CJ. Genetic diver-sity of Fusarium oxysporum f. sp. cubense causing Panama wilt of banana in the Philippines. Pathogens. 2019;9(1):32. https://doi.org/10.3390/pathogens9010032
  37. Aiello D, Carrieri R, Guarnaccia V, Vitale A, Lahoz E, Polizzi G. Characterization and pathogenicity of Colletotrichum gloeospori-oides and C. karstii causing preharvest disease on Citrus sinensis in Italy. Journal of Phytopathology. 2015;163(3):168-77. https://doi.org/10.1111/jph.12299
  38. Watanabe M, Yonezawa T, Lee KI, Kumagai S, Sugita-Konishi Y, Goto K, Hara-Kudo Y. Molecular phylogeny of the higher and lower taxonomy of the Fusarium genus and differences in the evolutionary histories of multiple genes. BMC Evolutionary Biol-ogy. 2011;11:1-6. https://doi.org/10.1186/1471-2148-11-322
  39. Castellá G, Cabañes FJ. Phylogenetic diversity of Fusarium incar-natum-equiseti species complex isolated from Spanish wheat. Antonie Van Leeuwenhoek. 2014;106:309-17. https://doi.org/10.1007/s10482-014-0200-x
  40. Maharaj A, Rampersad SN. Genetic differentiation of Colleto-trichum gloeosporioides and C. truncatum associated with an-thracnose disease of papaya (Carica papaya L.) and bell pepper (Capsicum annuum L.) based on ITS PCR-RFLP fingerprinting. Molecular Biotechnology. 2012;50:237-49. https://doi.org/10.1007/s12033-011-9434-2
  41. Mendy TK, Misran A, Mahmud TM, Ismail SI. Antifungal proper-ties of Aloe vera through in vitro and in vivo screening against postharvest pathogens of papaya fruit. Scientia Horticulturae. 2019;257:108767. https://doi.org/10.1016/j.scienta.2019.108767
  42. Bonde SR, Gade AK, Rai MK. Genetic variations among ten iso-lates of Fusarium equiseti (Corda) saccardo isolated from fruits and vegetables. Austin J Biotechnol Bioeng. 2014;1(5):5.
  43. Li YG, Song XL, Wang XQ, Zhang H, Tian S, Ji P. First report of fruit rot of watermelon caused by Fusarium equiseti in China. Plant Disease. 2018;102(9):1852. https://doi.org/10.1094/PDIS-01-18-0199-PDN
  44. Rahman MZ, Ahmad K, Siddiqui Y, Saad N, Hun TG, Mohd Hata E, et al. First report of Fusarium equiseti causing fruit rot disease of watermelon in Malaysia. Plant Disease. 2022;106(1):326. https://doi.org/10.1094/PDIS-05-21-1027-PDN
  45. Molnár O, Bartók T, Szécsi Á. Occurrence of Fusarium verticil-lioides and Fusarium musae on banana fruits marketed in Hun-gary. Acta Microbiologica et Immunologica Hungarica. 2015;62(2):109-19. https://doi.org/10.1556/030.62.2015.2.2
  46. Van Hove F, Waalwijk C, Logrieco A, Munaut F, Moretti A. Gibber-ella musae (Fusarium musae) sp. nov., a recently discovered species from banana is sister to F. verticillioides. Mycologia. 2011;103(3):570-85. https://doi.org/10.3852/10-038
  47. Wu KL, Chen WZ, Shuai YA, Ya WE, Zheng YR, Anjago WM, et al. Isolation and identification of Fusarium oxysporum f. sp. cubense in Fujian Province, China. Journal of Integrative Agriculture. 2019;18(8):1905-13. https://doi.org/10.1016/S2095-3119(18)62149-5

Downloads

Download data is not yet available.