Genetic variation of Colletotrichum spp. causing leaf anthracnose disease in citrus (Citrus spp.) on Java, Indonesia

  • Nabila Khairunnisa Salma
    Department of Plant Protection, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora No.1, Bulaksumur, Sleman, Yogyakarta, Indonesia 55581
  • Ani Widiastuti
    Department of Plant Protection, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora No.1, Bulaksumur, Sleman, Yogyakarta, Indonesia 55581
  • Khaerani Nurlaelita
    Department of Plant Protection, Faculty of Agriculture, Universitas Jember. Jl. Kalimantan No. 37, Jember, Indonesia 68121
  • Arif Wibowo
    Department of Plant Protection, Faculty of Agriculture, Universitas Gadjah Mada. Jl. Flora No.1, Bulaksumur, Sleman, Yogyakarta, Indonesia 55581

Abstract

Citrus is an important horticultural commodity in Indonesia, particularly for fresh consumption and as a raw material for processed products. Anthracnose caused by Colletotrichum spp. is an important disease of citrus that can substantially affect plant health and productivity. However, information on the diversity and identity of Colletotrichum species associated with citrus anthracnose in Java, Indonesia, remains limited. In this study, twelve Colletotrichum isolates were obtained from citrus leaves showing anthracnose symptoms collected from several citrus-growing regions in Java. The isolates were characterized based on morphological characteristics and genetic fingerprinting using repetitive element sequence-based PCR (rep-PCR) with BOX and ERIC primers. Seven representative isolates were subsequently selected for multilocus molecular identification and phylogenetic analysis based on partial sequences of five loci: internal transcribed spacer (ITS), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin (TUB2), and Apn2-Mat1-2 intergenic spacer (ApMat). The multilocus phylogenetic analysis placed the representative isolates into two major lineages corresponding to Colletotrichum siamense and the C. gloeosporioides species complex. Morphological characteristics were generally consistent with their molecular placement, although considerable overlap was observed among isolates. Pathogenicity assays on detached citrus leaves showed that all representative isolates produced anthracnose-like lesions, with differences in lesion development among isolates. These findings demonstrate the occurrence of genetically and phenotypically variable Colletotrichum isolates associated with citrus leaf anthracnose in Java and provide baseline information for further studies on pathogen diversity and disease management.

How to Cite

(1)
Genetic Variation of Colletotrichum Spp. Causing Leaf Anthracnose Disease in Citrus (Citrus spp.) on Java, Indonesia. J Trop Plant Pests Dis 2026, 26 (2), 602-614. https://doi.org/10.23960/j.hptt.226602-614.

References

Akin HM, Anggraini D, Wibowo L, Prasetyo J, & Suharjo R. 2024. Antifungal evaluation of turmeric rhizome extract against Colletotrichum capsici, the causal agent of anthracnose on red-chili peppers (Capsicum annuum L.). J. Trop. Plant Pests Dis. 24(1): 75–81. https://doi.org/10.23960/jhptt.12475-81
Anggrahini DS, Wibowo A, & Subandiyah S. 2020. Morphological and molecular identification of Colletotrichum spp. associated with chili anthracnose disease in Yogyakarta Region. J. Perlind. Tanam. Indones. 24(2): 161–174. https://doi.org/10.22146/jpti.58955
Benatar GV, Wibowo A, & Suryanti. 2021. First report of Colletotrichum asianum associated with mango fruit anthracnose in Indonesia. Crop Prot. 141: 105432. https://doi.org/10.1016/j.cropro.2020.105432
BPS-Statistic Indonesia. 2023. Production of Fruits. BPS Statistics Indonesia. https://www.bps.go.id/en/statistics-table/2/NjIjMg==/production-of-fruits.html. Accessed 12 August 2024.
Cai L, Hyde KD, Taylor PWJ, Weir B, Waller J, Abang MM, Zhang JZ, Yang YL, Phoulivong S, Liu ZY, Prihastuti H, Shivas RG, McKenzie EHC, & Johnston PR. 2009. A polyphasic approach for studying Colletotrichum. Fungal Divers. 39(1): 183–204. https://era.dpi.qld.gov.au/id/eprint/8483/1/FD39-8.pdf
Cannon PF, Damm U, Johnston PR, & Weir BS. 2012. Colletotrichum: Current status and future directions. Stud. Mycol. 73(1): 181–213. https://doi.org/10.3114/sim0014
Carbone I & Kohn LM. 1999. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia. 91(3): 553–556. https://doi.org/10.1080/00275514.1999.12061051
Choi YW, Hyde KD, & Ho WWH. 1999. Single spore isolation of fungi. Fungal Divers. 3: 29–38. https://fungaldiversity.org/fdp/sfdp/FD_3_29-38.pdf
De Silva DD, Crous PW, Ades PK, Hyde KD, & Taylor PWJ. 2017. Life styles of Colletotrichum species and implications for plant biosecurity. Fungal Biol. Rev. 31(3): 155–168. https://doi.org/10.1016/j.fbr.2017.05.001
Fayyaz A, Sahi ST, Nawaz-ul-Rehman MS, & Amrao L. 2020. Molecular characterization of fungal pathogens associated with citrus withertip/dieback from major citrus growing areas of Punjab, Pakistan. Int. J. Agric. Biol. 23(6): 1165–1170. https://doi.org/10.17957/IJAB/15.1400
Gautam AK, Verma RK, Avasthi S, Sushma, Bohra Y, Devadatha B, Niranjan M, & Suwannarach N. 2022. Current insight into traditional and modern methods in fungal diversity estimates. J. Fungi. 8(3): 226. https://doi.org/10.3390/jof8030226
Guarnaccia V, Groenewald JZ, Polizzi G, & Crous PW. 2017. High species diversity in Colletotrichum associated with citrus diseases in Europe. Persoonia. 39(1): 32–50. https://doi.org/10.3767/persoonia.2017.39.02
Guerber JC, Liu B, Correll JC, & Johnston PR. 2003. Characterization of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs, and mating compatibility. Mycological. 95(5): 872–895. https://doi.org/10.1080/15572536.2004.11833047
Masyahit M, Sijam K, Awang Y, & Satar MGM. 2009. The first report of the occurrence of anthracnose disease caused by Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. on dragon fruit (Hylocereus spp.) in Peninsular Malaysia. Am. J. Appl. Sci. 6(5): 902–912. https://doi.org/10.3844/ajassp.2009.902.912
Mo J, Zhao G, Li Q, Solangi GS, Tang L, Guo T, Huang S, & Hsiang T. 2018. Identification and characterization of Colletotrichum species associated with mango anthracnose in Guangxi, China. Plant Dis. 102(7): 1283–1289. https://doi.org/10.1094/PDIS-09-17-1516-RE
Nurlaelita K, Wibowo A, & Widiastuti A. 2024. Identification and characterization Colletotrichum spp. causing mango dieback in Indonesia. Plant Pathol. J. 40(4): 399–407. https://doi.org/10.5423/PPJ.NT.04.2024.0061
Patricia LCC, María del Socorro RG, Iván RH, Erika DlCA, Carolina DS, Keiko S, & José Alberto NZ. 2021. Occurrence and infective potential of Colletotrichum gloeosporioides isolates associated to Citrus limon var Eureka. Biotechnol. Rep. 31: e00651. https://doi.org/10.1016/j.btre.2021.e00651
Pramunadipta S, Widiastuti A, Wibowo A, & Priyatmojo A. 2021. Genetic diversity of the pathogenic fungus Sarocladium oryzae causing sheath rot on rice using rep-PCR. IOP Conf. Ser.: Earth Environ. Sci. 667: 012057. https://doi.org/10.1088/1755-1315/667/1/012057
Prihastuti H, Cai L, Chen H, McKenzie E, & Hyde KD. 2009. Characterization of Colletotrichum species associated with coffee berries in Northern Thailand. Fungal Divers. 39(1): 89–109.
Rhaiem A & Taylor PWJ. 2016. Colletotrichum gloeosporioides associated with anthracnose symptoms on citrus, a new report for Tunisia. Eur. J. Plant Pathol. 146(1): 219–224. https://doi.org/10.1007/s10658-016-0907-9
Sharma G, Pinnaka AK, & Shenoy BD. 2015. Resolving the Colletotrichum siamense species complex using ApMat marker. Fungal Divers. 71(1): 247–264. https://doi.org/10.1007/s13225-014-0312-7
Shidiq MAA, Widyaningsih S, Wibowo A, & Widiastuti A. 2024. First report of Colletotrichum queenslandicum and Colletotrichum endophyticum causing citrus anthracnose in Indonesia. J. Saudi Soc. Agric. Sci. 23(6): 393–403. https://doi.org/10.1016/j.jssas.2024.03.005
Silva DN, Talhinhas P, Várzea V, Cai L, Paulo OS, & Batista D. 2012. Application of the Apn2/MAT locus to improve the systematics of the Colletotrichum gloeosporioides complex: An example from coffee (Coffea spp.) hosts. Mycologia. 104(2): 396–409. https://doi.org/10.3852/11-145
Sudirga SK, Darmadi AAK, Wijaya IMS, & Yulihastuti DA. 2024. In vitro assessment of antifungal activity of cinnamon leaves extract against the Colletotrichum sp. causes of anthracnose on tomato. J. Trop. Plant Pests Dis. 24(2): 284–294. https://doi.org/10.23960/jhptt.224284-294
Wang W, de Silva DD, Moslemi A, Edrwards J, Ades PK, Crous PW, & Taylor PWJ. 2021. Colletotrichum species causing anthracnose of citrus in Australia. J. Fungi. 7(1): 47. https://doi.org/https://doi.org/ 10.3390/jof7010047
White TJ, Bruns T, Lee S, & Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. in PCR Protocols: a guide to methods and applications. pp. 315–322. Academic Press, Inc, California. https://doi.org/10.1016/b978-0-12-372180-8.50042-1
Woudenberg JHC, Aveskamp MM, de Gruyter J, Spiers AG, & Crous PW. 2009. Multiple Didymella teleomorphs are linked to the Phoma clematidina morphotype. Persoonia. 22(1): 56–62. https://doi.org/10.3767/003158509X427808