IN-VITRO ANTIFUNGAL POTENTIAL OF dsRNA MOLECULES ON FUSARIUM ACTIN RELATED PROTEIN 2/3, DNA POLYMERASE DELTA SUBUNIT AND ADENYLASE CYCLASE ESSENTIAL GENES ON COLLAR ROT AND WILT PATHOGENS OF PASSIONFRUIT.

Authors

  • Florence Nassimbwa School of Biosecurity, Biotechnical and Laboratory Sciences Makerere University. P. O. Box 7062 Kampala, Uganda.
  • Enock Matovu School of Biosecurity, Biotechnical and Laboratory Sciences Makerere University. P. O. Box 7062 Kampala, Uganda.
  • Andrew Kiggundu National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.
  • Charles Changa National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.
  • Francis Mumbanza National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.
  • John Adriko National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

DOI:

https://doi.org/10.51168/sjhrafrica.v4i6.363

Keywords:

Antifungal potential, actin related protein, adenlylase cyclase, dsRNA, PCR, polymerase delta subunit, RNAi

Abstract

BACKGROUND:

In Uganda two fungal diseases have become economically important; Fusarium wilt which is caused by.Fusarium oxysporum. f.sp passiflorae (Fop) and collar rot caused by Fusarium solani (Fs). The aim of this study was to determine antifungal activity of synthetic dsRNA constructed to silence essential genes; Actin Related Protein 2/3 (D6), DNA Polymerase delta subunit (J6), and Adenylase cyclase (K6), in Fusarium from banana on Fusarium sp from passionfruit.

METHODS:

In this study, pathogenic samples which were previously isolated from 3 districts in Central Uganda i.e. Wakiso, Mpigi, and Mukono were tested to determine and explore RNAi as an anti-fungal control strategy for spore germination. Sample materials obtained were subjected to DNA extraction from pure mycelia and PCR was performed. Antifungal dsRNA assays were run to test the efficiency relying on the reduction in a number of fungal colonies on PDA.

RESULTS:

Previous results showed that wilting was associated with one specific species of Fusarium oxysporum and collar rot with one species of Fusarium solani. Anti-fungal assay tests in this study included PCRs targeting the Fusarium actin related protein 2/3 (D6), DNA polymerase delta subunit (J6) as well as adenylase cyclase (K6) results showed that % spore germination inhibition on Fop ranged from 61.9-93.9% and Fs ranged from -69-87.3%.

CONCLUSION:

These results show that an environmentally friendly control measure for the above diseases is possible through the development of resistant plants which express dsRNA for a specified gene_ Fusarium Actin Related Protein 2/3.

RECOMMENDATION:

RNAi would be a promising target gene for RNAi –mediated resistance in passion fruit against the target fungal diseases.

Author Biographies

Florence Nassimbwa, School of Biosecurity, Biotechnical and Laboratory Sciences Makerere University. P. O. Box 7062 Kampala, Uganda.

(MSc. Molecular Biology) is a Biologist (Lecturer) and an instructor (FOO) at DAS Aviation School, Entebbe, and a Ph.D. candidate in Biological sciences at Kyambogo University. I have knowledge and experience in Biological sciences and Aviation Human Performance and Limitations(Human Factors).

Enock Matovu, School of Biosecurity, Biotechnical and Laboratory Sciences Makerere University. P. O. Box 7062 Kampala, Uganda.

Ass. Prof. Enock Matovu obtained his Ph.D. in Molecular Parasitology from the University of Bern, Switzerland in 2001, while he worked as a Research Officer at the Livestock Health Research Institute, Tororo, Uganda. Since then has continued his work on drug resistance and later diagnostics for African Trypanosomiasis. In 2004, he relocated to the Makerere University School of Veterinary Medicine, where he was first employed as a Lecturer. In 2008, Enock received the prestigious Royal Society Pfizer Award in recognition of his work on molecular mechanisms of drug resistance in African trypanosomes. The year (2007) he had obtained the Joint Third World Academy of Science Award for Young Scientists, for his contribution to the field of Molecular Parasitology. Enock Matovu has vast experience in HAT ranging from surveillance,diagnostics, drug resistance, and clinical trials.

Andrew Kiggundu, National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

Senior research scientist with National Agricultural Research Organization (NARO). University of Pretoria, South Africa. Forestry and Agricultural Biotechnology Institute (FABI) and the Department of Botany, Ph.D. in Plant Biotechnology.

Charles Changa, National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

Francis Mumbanza, National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

Researcher at the University of Ghent: Contact (voluntary lab member). CAVElab - Computational & Applied Vege- tation EcologyFrancis Mumbanza Mundondo Department of Environment, Faculty of Bioscience Engineering.

John Adriko, National Biotechnology Research Center/ Tissue Culture, National Agricultural Research Laboratories. P. O. Box 7065 Kampala, Uganda.

Is a Crop Scientist with experience in both field agriculture and research. Dr. Adriko holds a Ph.D. in Plant Pathology from the University of Copenhagen, a Masters’s Degree in Crop Science, and a bachelor’s Degree in Agriculture, both from Makerere University. He has valuable skills and experience in agriculture and food security obtained during his work with District Local Government Production Department, National Agricultural Advisory Services (NAADS), and National Agricultural and Research Organization (NARO). He also worked as Agricultural Consultant for NilePro Consult Limited and is now Arise Development Consultant playing a key role in training farmers on agriculture production for income and food security for the Arua District Local Government, World Vision, CARE Uganda, and Eastern Archdiocesan Development Network (EADEN). He also worked as Field Extension Agent for Farm Talk/ Tree Talk in West Nile (Straight Talk Foundation). He has carried out farmer training in improved crop production for Governmental as well as Non-governmental institutions. Dr. Adriko was the agriculture technical expert in a market access survey done for ZOA Uganda and the Lead Consultant in agriculture,food security, and child nutrition analysis for Plan Uganda.

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Published

2023-06-29

How to Cite

Nassimbwa, F., Matovu, E. ., Kiggundu, A. ., Changa, C. ., Mumbanza, F. ., & Adriko, J. (2023). IN-VITRO ANTIFUNGAL POTENTIAL OF dsRNA MOLECULES ON FUSARIUM ACTIN RELATED PROTEIN 2/3, DNA POLYMERASE DELTA SUBUNIT AND ADENYLASE CYCLASE ESSENTIAL GENES ON COLLAR ROT AND WILT PATHOGENS OF PASSIONFRUIT. Student’s Journal of Health Research Africa, 4(6), 9. https://doi.org/10.51168/sjhrafrica.v4i6.363

Issue

Vol. 4 No. 6 (2023): June 2023 Issue

Section

Section of Genetics, and Biotechnology

License

Copyright (c) 2023 Florence Nassimbwa, Enock Matovu, Andrew Kiggundu, Charles Changa, Francis Mumbanza, John Adriko

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.