Silva, Goncalo ORCID: 0000-0001-5544-2947 , Bömer, Moritz ORCID: 0000-0003-1003-9622 , Turaki, Aliyu A., Nkere, Chukwuemeka K., Kumar, P. Lava and Seal, Susan E. ORCID: 0000-0002-3952-1562 (2022) Homing in on endogenous badnaviral elements: development of multiplex PCR-DGGE for detection and rapid identification of badnavirus sequences in yam germplasm. Frontiers in Plant Science, 13:846989. ISSN 1664-462X (Online) (doi:https://doi.org/10.3389/fpls.2022.846989)

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Abstract

Viruses of the genus Badnavirus (family Caulimoviridae) are double-stranded DNA-reverse transcribing (dsDNA-RT) plant viruses and have emerged as serious pathogens of tropical and temperate crops globally. Endogenous badnaviral sequences are found integrated in the genomes of several economically important plant species. Infection due to activation of replication-competent integrated copies of the genera Badnavirus, Petuvirus and Cavemovirus has been described. Such endogenous badnaviral elements pose challenges to the development of nucleic acid-based diagnostic methods for episomal virus infections and decisions on health certification for international movement of germplasm and seed. One major food security crop affected is yam (Dioscorea spp.). A diverse range of Dioscorea bacilliform viruses (DBVs), and endogenous DBV (eDBV) sequences have been found to be widespread in yams cultivated in West Africa and other parts of the world. This study outlines the development of multiplex PCR-dependent denaturing gradient gel electrophoresis (PCR-DGGE) to assist in the detection and analysis of eDBVs, through the example of analysing yam germplasm from Nigeria and Ghana. Primers targeting the three most prevalent DBV monophyletic species groups in West Africa were designed to improve DGGE resolution of complex eDBV sequence fingerprints. Multiplex PCR-DGGE with the addition of a tailor-made DGGE sequence marker enables rapid comparison of endogenous badnaviral sequence diversity across germplasm, as illustrated in this study for eDBV diversity in yam.

Item Type:	Article
Additional Information:	THIS ARTICLE IS PART OF THE RESEARCH TOPIC "DNA Virus and Host Plant Interactions from Antagonism to Endogenization".
Uncontrolled Keywords:	badnavirus; multiplex PCR; denaturing gradient gel electrophoresis (DGGE); endogenous badnaviral elements; yam; Dioscorea spp.; West Africa
Subjects:	Q Science > QR Microbiology > QR355 Virology S Agriculture > S Agriculture (General) S Agriculture > SB Plant culture
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Engineering & Science > Natural Resources Institute Faculty of Engineering & Science > Natural Resources Institute > Agriculture, Health & Environment Department Faculty of Engineering & Science > Natural Resources Institute > Molecular Virology and Entomology Research Group
Last Modified:	11 May 2022 08:53
URI:	http://gala.gre.ac.uk/id/eprint/35947

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