Journal of Applied & Environmental Microbiology
ISSN (Print): 2373-6747 ISSN (Online): 2373-6712 Website: Editor-in-chief: Sankar Narayan Sinha
Open Access
Journal Browser
Journal of Applied & Environmental Microbiology. 2020, 8(1), 25-31
DOI: 10.12691/jaem-8-1-4
Open AccessArticle

Screening and Selection in vitro and in vivo of Cocoa Tree (Theobroma Cacao Linn) Endophytic Bacteria Having Antagonistic Effects against Phytophthora Spp. Fungal Agents Responsible of Black Pod Disease in Côte d’Ivoire

OUATTARA Adama1, COULIBALY Klotioloma2, KONATE Ibrahim1, , KEBE Boubacar Ismaël2, BEUGRE G.A. Maxwell1, TIDOU Abiba Sanogo1 and Abdelkarim FILALI-MALTOUF3

1UFR Agro-forestry, Department of Biochemistry and Microbiology, Laboratory of Agrovalorization, Jean Lorougnon Guede University, B.P: 150 Daloa, Côte d’Ivoire

2National Center for Agronomic Research (CNRA), Laboratory of Phytopathology, BP: 808 Divo, Côte d’Ivoire

3Faculty of Sciences, Laboratory of Microbiology and Molecular Biology, University Mohammed V-Agdal, Rabat, Morocco

Pub. Date: March 29, 2020

Cite this paper:
OUATTARA Adama, COULIBALY Klotioloma, KONATE Ibrahim, KEBE Boubacar Ismaël, BEUGRE G.A. Maxwell, TIDOU Abiba Sanogo and Abdelkarim FILALI-MALTOUF. Screening and Selection in vitro and in vivo of Cocoa Tree (Theobroma Cacao Linn) Endophytic Bacteria Having Antagonistic Effects against Phytophthora Spp. Fungal Agents Responsible of Black Pod Disease in Côte d’Ivoire. Journal of Applied & Environmental Microbiology. 2020; 8(1):25-31. doi: 10.12691/jaem-8-1-4


We are investigating in this study the screening and selection of cocoa tree (Theobroma cacao L.) endophytic bacteria having antagonistic effects against the fungi Phytophthora spp. and to assess their inhibitory potential as biological control agents against black pod disease in Côte d'Ivoire. A total of One hundred and sixteen bacteria were isolated from symptomless root, leaf and stem tissues of two cocoa clones NA32 (susceptible) and P7 (resistant) young seedlings. These isolates restricted the radial growth of the mycelium of P. palmivora (strain BL7.11.2) and P. megakarya (strain 13P30.1) with percentages inhibition of 28.58 to 70.54% and 25.3 to 64.29% respectively. Four isolates, 48P, 60P, 23P and 18N were more effective in in vitro test and selected for in vivo confrontation test on leaf discs. Treatments based on two isolates S1 (48P) and S2 (18N) applied at the concentration 109 CFU/mL significantly (p < 0.05) reduced the black pod necrosis induced by the two Phytophthora strains on the leaf discs of three cocoa clones (NA32, PA150 and SCA6). In addition, simultaneous (C3PS1 and C3PS2) or separate (C3AVS1 and C3AVS2) inoculations of these two bacteria and Phytophthora, induced resistance to the clone NA32 and increased the intrinsic resistance of the clones PA150 and SCA6. This study showed that endophytic bacteria antagonistic to Phytophthora spp. can be an alternative solution in the biocontrol of black pod disease. In situ pod tests should be performed to confirm these preliminary results and propose effective bacterial inoculum.

black pod phytophthora spp endophytic bacteria cocoa tree biocontrol

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Ploetz R. The Impact of Diseases on Cacao Production: A Global Overview in: B.A. Bailey, L.W. Meinhardt (eds.), Cacao Diseases, Springer International Publishing Switzerland.p33-59. 2016.
[2]  Coulibaly K., Aka R. A., Camara B., Kassin E., Kouakou K., Kébé B. I., Koffi N. K., Tahi M. G., Walet N. P., Guiraud S. B., Assi M. E., Kone B., N’Guessan K. F, Koné D.. Molecular Identification of Phytophthora palmivora in the Cocoa Tree Orchard of Côte d’Ivoire and Assessment of the Quantitative Component of Pathogenicity. Internationale journale of science, 7 (8). 7-15. August 2018.
[3]  Mpika Joseph, Kebe Ismaël B., Druzhinina Irina S., Komon-Zélazowska Monika, Kubicek Christian P. & Aké Sevérin. Inhibition de Phytophthora palmivora, agent de pourriture brune des cabosses de cacaoyer en Côte d’Ivoire, par Trichoderma sp. Sciences & Nature, 6(1). 49-62. Mars 2009.
[4]  Mbargaa J.B., Begoude B.A.D., Ambang Z., Meboma M., Kuate J., Schiffers B., Ewbankd W., Dedieu L., ten Hoopen G.M. A new oil-based formulation of Trichoderma asperellum for the biological control of cacao black pod disease caused by Phytophthora megakarya. Biological Control, 77. 15-22. June 2014.
[5]  Pohe Jean, Pohe Sédjouény Steve Wilfrid, Okou Staelle Florence Famisso. Association oxyde de cuivre et metalaxyl dans la lutte contre la pourriture brune des cabosses de cacaoyer en Côte d’Ivoire. Journal of Animal &Plant Sciences, 16(3). 2362-2368. January 2013.
[6]  Iwaro A.D., Thévenin J.-M., Butler D.R. and Eskes A.B. Usefulness of the detached pod test for assessment of cacao resistance to Phytophthora pod rot. European Journal of Plant Pathology, 113. 173-182. August 2005.
[7]  Tahi, G.M., Kébé, B.I., N’Goran, A.K.N., Sangare´, A., Mondeil, F., Cilas, C., Eskes, A.B. Expected selection efficiency for resistance to cacao pod rot (Phytophthora palmivora) comparing lead disc inoculations with field observations. Euphytica 149, 35-44. November 2006.
[8]  Tjamos, E.C., Tjamos, S.E. & Antoniou, P.P. Biological management of plant diseases: Highlights on research and application. Journal of Plant Pathology, 92(4). 17-21. May 2010.
[9]  Soylu, S., Soylu, E.M., Kurt, S. & Ekici, O.K. Antagonistic potentials of rhizosphere-associated bacterial isolates against soil-borne diseases of tomato and pepper caused by Sclerotinia sclerotiorum and Rhizoctonia solani. Pakistan Journal of Biological Sciences, 8(1). 43-48. 2005.
[10]  Nur Rashyeda Ramli, Maizatul Suriza Mohamed, Idris Abu Seman, Madihah Ahmad Zairun & Nasyaruddin MohamadThe Potential of Endophytic Bacteria as a Biological Control Agent for Ganoderma Disease in Oil Palm. Sains Malaysiana, 45(3). 401-409. 2016.
[11]  Ouattara Adama, Coulibaly Klotioloma, Konate Ibrahim, Kebe Boubacar Ismaël, Tidou Abiba Sanogo, Filali-Maltouf Abdelkarim. Selection of Cocoa Tree (Theobroma cacao Linn) Endophytic Bacteria Solubilizing Tri-Calcium Phosphate, Isolated from Seedlings Grown on Soils of Six Producing Regions of Côte d’Ivoire. Advances in Microbiology, 9. 842-852. September 2019.
[12]  Melnick Rachel L., Suárez Carmen, Bailey Bryan A., Backman Paul A. Isolation of endophytic endospore-forming bacteria from Theobroma cacao as potential biological control agents of cacao diseases. Biological Control, 57. 236-245. March 2011.
[13]  Abdallah Aydi-Ben Rania, Marwa Hassine, Hayfa Jabnoun-Khiareddine, Hichem Ben Jannet & Mejda Daami-Remadi. Etude du pouvoir antifongique in vitro et in vivo des filtrats de culture et des extraits organiques des Aspergillus spp. contre Phytophthora sp. Microbiol. Hyg. Alim, 25(73).15-25. July 2013.
[14]  Rika Fithri, N.B., Aris Tri, W. & Nurita, T.M. Control activity of potential antifungal-producing Burkholderia sp. in suppressing Ganoderma boninense growth in oil palm. Asian Journal of Agricultural Research, 8. 259-268. 2014.
[15]  Konate, I., Ouattara, A., Coulibaly, B., Guei, N.K.R., Amani, K., Kouakou, I.K., Filali-Maltouf, A. and Koffi, M. Phenotypic Diversity of Associative Bacteria Isolated from Roots and Stems of Cacao (Theorem cacao) Tree in Daloa, Côte d’Ivoire. International Journal of Current Microbiology and Applied Sciences, 4. 560-570. August 2015.
[16]  Rayda S., Ines B. C., Tatiana V., Mohamed A. T., Mouna J., Manel C., Imen A., Amine E., Ahmad N., Frederique C., Radhouane G., Noureddine D., and Néji G. Analysis of the Cultivable Endophytic Bacterial Diversity in the Date Palm (Phoenix dactylifera L.) and Evaluation of Its Antagonistic Potential against Pathogenic Fusarium Species that Cause Date Palm Bayound Disease. Journal of Applied & Environmental Microbiology, 4(5). 93-104. September 2016.
[17]  Melnick Rachel L., Zidack Nina K., Bailey Bryan A., Maximova Siela N., Guiltinan Mark, Backman Paul A. Bacterial endophytes: Bacillus spp. from annual crops as potential biological control agents of black pod rot of cacao. Biological Control, 46. 46–56. March 2008.
[18]  Akrofi A. Y., Terlabie J. L., Amoako-Attah I., Asare E. K.. Isolation and characterization of bacteria from different cacao progenies and their antagonistic activity against the black pod disease pathogen, Phytophthora palmivora. Journal of Plant Dis Prot, 124.143-152. March 2017.
[19]  Nyassé S., Cilas C., Herail C. and Blaha G. Leaf inoculation as an early screening test for cocoa (Theobroma cacao L.) resistance to Phytophthora black pod disease. Crop Protection, 14(8). 657-663. January 1995.
[20]  Kébé Ismaël B., Mpika Joseph, N’Guessan Kouamé F., Hebbar Prakash K., Samuels Gary S. & AKE Severin. Isolement et identification de microorganismes indigènes de cacaoyères en Côte d’Ivoire et mise en évidence de leurs effets antagonistes vis-à-vis de Phytophthora palmivora, agent de la pourriture brune des cabosses. Sciences & Nature, 6 (1). 71-82. Avril 2009.
[21]  SAS. Guide to the use of PC-SAS Version 6.04 for DOS for Statistical Analysis. SAS Institute, Cary, North Carolina. 1995.
[22]  Gomez, K.A. & Gomez, A.A. Statistical Procedures for Agricultural Research. 2nd ed. Chapter 2. New York: John Wiley & Sons. pp. 7-83. 1984.
[23]  Acebo-Guerrero1 Y., Hernandez-Rodrıguez A., Vandeputte O., Miguelez-Sierra Y., Heydrich-Perez M., Ye L., Cornelis P., Bertin P and El Jaziri M. Characterization of Pseudomonas chlororaphis from Theobroma cacao L. rhizosphere with antagonistic activity against Phytophthora palmivora (Butler). Journal of Applied Microbiology, 119. 1112-1126. July 2015.
[24]  Mejia Luis C., Rojas Enith I., Maynard Zuleyka., Bael Sunshine Van, Arnold A. Elizabeth, Hebbar Prakash., Samuels Gary J., Robbins Nancy, Herre Edward Allen. Endophytic fungi as biocontrol agents of Theobroma cacao pathogens. Biological Control. January 2008.