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PCR‐based identification of cacao black pod causal agents and identification of biological factors possibly contributing to Phytophthora megakarya's field dominance in West Africa
Authors:S. S. Ali  I. Amoako‐Attah  R. A. Bailey  M. D. Strem  M. Schmidt  A. Y. Akrofi  S. Surujdeo‐Maharaj  O. O. Kolawole  B. A. D. Begoude  G. M. ten Hoopen  E. Goss  W. Phillips‐Mora  L. W. Meinhardt  B. A. Bailey
Affiliation:1. Sustainable Perennial Crops Laboratory, Plant Sciences Institute, USDA/ARS, Beltsville Agricultural Research Center‐West, Beltsville, MD, USA;2. Cocoa Research Institute of Ghana, Akim New‐Tafo, Ghana;3. Cocoa Research Centre, the University of the West Indies, St Augustine, Trinidad and Tobago;4. Cocoa Research Institute of Nigeria, Ibadan, Nigeria;5. Plant Pathology Department, University of Florida, Gainesville, FL, USA;6. Regional Laboratory for Biological and Applied Microbiology, IRAD, Yaoundé, Cameroon;7. CIRAD, UPR 106 Bioagresseurs, Montpellier, France;8. Department of Agriculture and Agroforestry, CATIE, Turrialba, Costa Rica
Abstract:Among the Phytophthora species that cause black pod of cacao, P. megakarya is the most virulent, posing a serious threat to cacao production in Africa. Correct identification of the species causing the black pod and understanding the virulence factors involved are important for developing sustainable disease management strategies. A simple PCR‐based species identification method was developed using the species‐specific sequences in the ITS regions of the rRNA gene. A phylogenetic tree generated for 119 Phytophthora isolates, based on the 60S ribosomal protein L10 gene and rDNA sequence, verified the PCR‐based identification assay and showed high interspecific variation among the species causing black pod. Phytophthora megakarya isolates were uniformly virulent in an assay using susceptible cacao pod husks inoculated with zoospores, while the P. palmivora isolates showed greater divergence in virulence. The virulence of P. megakarya was associated with earlier production of sporangia and an accelerated induction of necrosis. While zoospore germ tubes of both species penetrated pods through stomata, only P. megakarya produced significant numbers of appressoria. A hypersensitive‐like response was observed when attached SCA‐6 pods were inoculated with P. palmivora. SCA‐6 pods became vulnerable to P. palmivora when wounded prior to zoospore inoculation. Phytophthora megakarya was more aggressive than P. palmivora on attached SCA‐6 pods, causing expanding necrotic lesions with or without wounding. Phytophthora megakarya is predominant in the Volta region of Ghana and it remains to be seen whether it can displace P. palmivora from cacao plantations of Ghana as it has in Nigeria and Cameroon.
Keywords:appressorial  cocoa  Ghana     PCR     phylogeny  virulence
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