Morphological and Molecular Characterization of Phytophthora palmivora Isolates and Their Virulence Test on Cocoa Clone Sulawesi 2
Abstract
Morphological characterization of Phytophthora isolates commonly was done on the basis of the colony and hyphal swelling characteristics, form, and shape of chlamydospore, sporangia as well as oogonia and antheridia. Molecular characterization was conducted using specific primer pairs Pal1 s and Pal2a for Phytophthora palmivora and universal primer ITS4 and ITS5 followed by sequencing of isolates. The virulence of Phytophthora palmivora isolates was tested on a cocoa pod of the Sulawesi2 clone. A total of 21 Phytophthora isolates were collected from Palopo, Gowa, and Pinrang Districts. Based on colony morphology and microscopic features i.e. chlamydospore, sporangium, and its papilla form, all isolates indicated Phytophthora palmivora. Molecular identification using a specific primer showed that all ten representative group isolates had the same band in size of 650 bp, whereas using universal ITS4 and ITS5 primers, all isolates generated one band in the size of 900bp. The sequencing result showed, that all isolates were placed in the group P. palmivora. The virulence of P. palmivora isolates on the Sulawesi2 clone revealed a varying degree of virulence, from less virulent to highly virulent. All isolates from Pinrang were classified as highly virulent because induced necrotic in more than 50% of the tested cocoa pod.
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Andrews YA, Ishmael AA, Michael A, Eric KA. 2015. Black pod disease on cocoa (Theobroma cocoa, L.) in Ghana: spread of Phytophthora megakarya and role of economic plants in the disease epidemiology. Crop Protection. 72:66–75. DOI: https://doi.org/10.1016/j.cropro.2015.01.015.
Appiah A, Flood J, Bridge PD, Archer SA. 1999. Diversity of Phytophthora species causing black pod disease of cocoa and implications for effective biocontrol. In: Krauss U, Hebbar P, Editor. Research methodology in biocontrol of plant diseases with special reference to fungal diseases of cocoa. In: Workshop on Research Methodology in Biocontrol of Plant Diseases with Special Reference to Fungal Diseases of Cocoa, 1999 28 Jun-4 Jul. Washington, DC (US): Agricultural Research Service.
Appiah A, Flood J, Bridge PD, Archer SA. 2003. Inter- and intraspecific morphometric variation and characterization of Phytophthora isolate from cocoa. Plant Pathology. 52(2):168–180. DOI: https://doi.org/10.1046/j.1365-3059.2003.00820.x.
Bowers JH, Bailey BA, Hebbar PK, Sanogo S, Lumsden RD. 2001. The impact of plant diseases on world chocolate production. Plant Health Progress. 2(1):12. DOI: https://doi.org/10.1094/PHP-2001-0709-01-RV.
Blair JE, Michael D. Coffey, Sook-Young Park, David M. Geiser, Seogchan Kang. 2008. A multi-locus phylogeny for Phytophthora utilizing markers derived from complete genome sequences. Fungal Genetics and Biology. 45(3):266–277. DOI: https://doi.org/10.1016/j.fgb.2007.10.010.
Capote N, Mara A, Aguado A, Snchez-Torres P. 2012. Molecular tools for detection of plant pathogenic fungi and fungicide resistance. Plant Pathology. 7:151–202. DOI: https://doi.org/10.5772/38011.
Chirapongsatonkul N, Chanwun T, Churngchow N. 2015. Development of a multiplex PCR assay for rapid and simultaneous detection of rubber tree pathogens Phytophthora spp. and P. palmivora. Science Asia. 41(3):170−179. DOI: https://doi.org/10.2306/scienceasia1513-1874.2015.41.170.
Cooke DEL, Drenth A, Duncan JM, Wagels G, Brasier CM. 2000. A molecular phylogeny of Phytophthora and related oomycetes. Fungal Genetics and Biology. 30:17–32. DOI: https://doi.org/10.1006/fgbi.2000.1202.
Doyle JJ, Doyle JL. 1990. Isolation of plant DNA from fresh tissue. Focus. 12:13–15. DOI: https://doi.org/10.2307/2419362.
Drenth A, Sendall B. 2001. Practical guide to detection and identification of Phytophthora. Tropical Plant Protection. 1:32–33.
Erwin DC, Ribeiro OK. 1996. Phytophthora Diseases Worldwide. St. Paul (US): APS Press. DOI: https://doi.org/10.1046/j.1365-3059.1998.0179a.x.
Fuzitani EJ, Santos AF, Damatto Junior ER, Nomura ES, Kalil Filho AN. 2018. Inoculation methods, the aggressiveness of isolates, and resistance of peach palm progenies to Phytophthora palmivora. Summa Phytopathologica. 44(3):213–217. DOI: https://doi.org/10.1590/0100-5405/178476.
Guest D. 2007. Black pod: diverse pathogens with a global impact on cocoa yield. Phytopathology. 97:1650–1653. DOI: https://doi.org/10.1094/PHYTO-97-12-1650.
Kaosiri T, Zentmyer GA, Erwin DC. 1978. Stalk length as a taxonomic criterion for Phytophthora palmivora isolates from cocoa. Canadian Journal of Botany. 56(15):1730–1738. DOI: https://doi.org/10.1139/b78-205.
Kroon LP, Brouwer H, de Cock AW, Govers F. 2012. The genus Phytophthora anno 2012. Phytopathology. 102(4):348–364. DOI: https://doi.org/10.1094/PHYTO-01-11-0025.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution. 35(6):1547–1549. DOI: https://doi.org/10.1093/molbev/msy096.
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD. 2007. Clustal W and clustal X version 2.0. Bioinformatics. 23(21):2947–2948. DOI: https://doi.org/10.1093/bioinformatics/btm404.
Martin FN, Abed ZG, Baldi Y, Ivors K. 2012. Identification and detection of Phytophthora: reviewing our progress, identifying our needs. Plant Disease. 96:1080–1103. DOI: https://doi.org/10.1094/PDIS-12-11-1036-FE.
Ristaino JB, Michael Madritch, Carol L. Trout, Gregory Parra. 1998. PCR amplification of ribosomal DNA for species identification in the plant pathogen genus Phytophthora. Applied and Environmental Microbiology. 64(3):948–954. DOI: https://doi.org/10.1128/AEM.64.3.948-954.1998.
Rodríguez-Polanco E, Morales JG, Muñoz-Agudelo M, Segura JD, Carrero ML. 2020. Morphological, molecular, and pathogenic characterization of Phytophthora palmivora isolates causing black pod rot of cocoa in Colombia. Spanish Journal of Agricultural Research. 18(2):1003–1018. DOI: https://doi.org/10.5424/sjar/2020182-15147.
Rubiyo A, Purwantara S, Sukamto, Sudarsono. 2008. Isolation of indigenous Phytophthora palmivora from Indonesia, their morphological and virulence characterizations. Pelita Perkebunan. 24:27–49. DOI: https://doi.org/10.22302/iccri.jur.pelitaperkebunan.v24i1.93.
Rubiyo YA, Dewi, Imran, Salim A, Baharudin, Indrawanto C, MT Ratule. 2020. Evaluation of yield and pest and disease resistance of cocoa clones in Kolaka District, Southeast Sulawesi, Indonesia. Biodiversitas. 21(12):5698–5607. DOI: https://doi.org/10.13057/biodiv/d211215.
Saul Maora J, Liew ECY, Guest DI. 2017. Limited morphological, physiological and genetic diversity of Phytophthora palmivora from cocoa in Papua New Guinea. Plant Pathology. 66:124–130. DOI: https://doi.org/10.1111/ppa.12557.
Scott PM, Burgess TI, Barber PA. 2009. Phytophthora multicore sp. nov., a new species recovered from declining eucalyptus, banksia, agonis, and other plant species in Western Australia. Persoonia. 22:1–13. DOI: https://doi.org/10.3767/003158509X415450.
Stamp DJ, Waterhouse GM, Newhook FJ, Hall GS. 1990. Revised Tabular Key to The Species of Phytophthora. Wallingford (UK): CAB-International.
Surujdeo-Maharaj S, Sreenivasan TN, Motilal LA, Umaharan P. 2016. Black pod and other Phytophthora induced diseases of cocoa: history, biology, and control. In: Bailey BA, Meinhardt L, editor. Cocoa Diseases: A History of Old Enemies and New Encounters, First edition. Switzerland (CH): Springer. pp. 213–266. DOI: https://doi.org/10.1007/978-3-319-24789-2_7.
Torres-Londoño GA. 2016. Morphological characterization, virulence, and fungicide sensitivity evaluation of Phytophthora palmivora [dissertation]. Michigan State University.
Trout CL, Ristaino JB, Madritch M, Wangsoomboondee T. 1997. Rapid detection of Phytophthora infestans in late blight infected potatoes and tomatoes using PCR. Plant Disease. 81:1042–1048. DOI: https://doi.org/10.1094/PDIS.1997.81.9.1042.
Umayah A, Purwantara A. 2006. Identifikasi isolat Phytophthora asal kakao. Menara Perkebunan. 74(2):76–85.
Wael Alsultan, Yun WM, Zainal Abidin Mior Ahmad ZAM, Vadamalai G, Khairulmazmi A, Abdullah M. Al-Sadi, Rashed O, Nasehi A, Al-Tawaha AR. (2022) Morphological, pathogenic, and molecular characterization of Phytophthora palmivora isolates causing black pod disease on cocoa in Peninsular Malaysia. International Journal of Food Science and Agriculture. 6(2):135–148. DOI: https://doi.org/10.26855/ijfsa.2022.06.002.
Waterhouse GM. 1974a. Phytophthora palmivora and some related species. In: Gregory PH, editor. Phytophthora Disease of Cocoa. London, Longman (UK): Cornell University. p. 51–70.
Waterhouse GM. 1974b. Other Phytophthora species recorded on Cocoa. In: Gregory PH, editor. Phytophthora Disease of Cocoa. London, Longman (UK): Cornell University. p. 71–79.
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