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Int. J. Mol. Sci. 2017, 18(11), 2327; https://doi.org/10.3390/ijms18112327

Genome Analysis and Genetic Stability of the Cryptophlebia leucotreta Granulovirus (CrleGV-SA) after 15 Years of Commercial Use as a Biopesticide

1
Department of Biochemistry and Microbiology, P.O. Box 94, Rhodes University, Grahamstown 6140, South Africa
2
Department of Recombinant Vaccines, Intercollegiate Faculty of Biotechnology University of Gdansk and Medical University of Gdansk, 80-210 Gdansk, Poland
3
River Bioscience, P.O. Box 20388, Humewood, Port Elizabeth 6013, South Africa
4
Citrus Research International, P.O. Box 20285, Humewood, Port Elizabeth 6013, South Africa
5
Department of Zoology and Entomology, P.O. Box 94, Rhodes University, Grahamstown 6140, South Africa
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 14 September 2017 / Revised: 27 October 2017 / Accepted: 30 October 2017 / Published: 3 November 2017
(This article belongs to the Special Issue Molecular Entomology of Insects of Economic Importance)
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Abstract

Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is an indigenous pest in southern Africa which attacks citrus fruits and other crops. To control T. leucotreta in South Africa, an integrated pest management (IPM) programme incorporating the baculovirus Cryptophlebia leucotreta granulovirus (CrleGV-SA) as a biopesticide has been implemented. This study investigated the genetic stability of a commercially produced CrleGV-SA product that has been applied in the field since 2000. Seven representative full-genome sequences of the CrleGV-SA isolate spanning a 15-year period were generated and compared with one another. Several open reading frames (ORFs) were identified to have acquired single nucleotide polymorphisms (SNPs) during the 15-year period, with three patterns observed and referred to as “stable”, “reversion”, and “unstable switching”. Three insertion events were also identified, two of which occurred within ORFs. Pairwise multiple alignments of these sequences showed an identity ranging from 99.98% to 99.99%. Concentration-response bioassays comparing samples of CrleGV-SA from 2000 and 2015 showed an increase in virulence toward neonate T. leucotreta larvae. The CrleGV-SA genome sequence generated from the 2015 sample was compared to the Cape Verde reference genome, CrleGV-CV3. Several fusion events were identified between ORFs within these genomes. These sequences shared 96.7% pairwise identity, confirming that CrleGV-SA is a genetically distinct isolate. The results of this study indicate that the genome of CrleGV-SA has remained stable over many years, with implications for its continued use as a biopesticide in the field. Furthermore, the study describes the first complete baculovirus genome to be sequenced with the MinION (Oxford Nanopore, Oxford, UK) platform and the first complete genome sequence of the South African CrleGV isolate. View Full-Text
Keywords: betabaculovirus; false codling moth; biopesticide; genome stability betabaculovirus; false codling moth; biopesticide; genome stability
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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van der Merwe, M.; Jukes, M.D.; Rabalski, L.; Knox, C.; Opoku-Debrah, J.K.; Moore, S.D.; Krejmer-Rabalska, M.; Szewczyk, B.; Hill, M.P. Genome Analysis and Genetic Stability of the Cryptophlebia leucotreta Granulovirus (CrleGV-SA) after 15 Years of Commercial Use as a Biopesticide. Int. J. Mol. Sci. 2017, 18, 2327.

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