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Non-destructive DNA extraction from aphids: the application in virus - vector studies of Banana bunchy top virus (BBTV)

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Abstract

Banana bunchy top virus (BBTV), one of the most devastating viruses of banana, is transmitted by the banana aphid, Pentalonia nigronervosa. The high degree of morphological similarity between P. nigronervosa and the related experimental vector P. caladii makes it difficult to distinguish the two species. DNA barcoding can be used as an alternative for the rapid identification of these aphid species. However, standard molecular identification techniques for small insects usually require maceration of the sample to obtain sufficient amounts of DNA, resulting in the loss of voucher specimens. In this study, a non-destructive DNA extraction method for aphids was optimised, allowing the specimens to remain intact for use as vouchers for further morphological studies. Sufficient DNA concentrations were obtained from the aphid extractions and were used for species identification through direct sequencing of PCR products. Extracted DNA was further used to detect BBTV in the insect vector, P. nigronervosa. Approximately 78% of the aphids collected from symptomatic plants in infected plantations in KwaZulu-Natal tested positive for BBTV, as well as some aphids collected in other African countries. The BBTV viral strain identified from South Africa grouped with the “South Pacific” clade. This non-destructive DNA extraction method can be used in an early detection management strategy, especially in epidemiology and virus-vector studies of Banana bunchy top disease (BBTD).

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Acknowledgements

As authors we would like to thank our partners in the Learning Alliance Network, at the University of Kisangani, for collecting and donating samples from outside Kenya and Uganda. The Professional Development Programme (PDP, Agricultural Research Council) and the University of Pretoria is acknowledged for support of N. Robbertse during her studies. We thank Janine Snyman (University of Pretoria) for identification of aphids collected in traps.

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Authors and Affiliations

  1. Plant Microbiology Division, Agricultural Research Council-Plant Protection Research (ARC-PPR), Pretoria, South Africa

    N. Robbertse & A. E. C. Jooste

  2. Department of Zoology and Entomology, University of Pretoria, Private Bag X20, Pretoria, 0028, South Africa

    N. Robbertse & K. Krüger

  3. Bioversity International, Katalima Road, Nagulu, Kampala, Uganda

    B. A. Omondi

  4. Biosystematics Division, Agricultural Research Council-Plant Protection Research (ARC-PPR), Pretoria, South Africa

    I. M. Millar

  5. Crop Protection Division, Agricultural Research Council - Tropical and Subtropical Crops (ARC-TSC), Nelspruit, South Africa

    A. E. C. Jooste

Authors
  1. N. Robbertse
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  2. B. A. Omondi
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  3. I. M. Millar
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  4. K. Krüger
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  5. A. E. C. Jooste
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Correspondence to A. E. C. Jooste.

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We hereby state that no conflict of interest are included in this manuscript and that no humans or animals were used in the study. All authors agreed to the publishing of this manuscript.

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Robbertse, N., Omondi, B.A., Millar, I.M. et al. Non-destructive DNA extraction from aphids: the application in virus - vector studies of Banana bunchy top virus (BBTV). Eur J Plant Pathol 153, 571–582 (2019). https://doi.org/10.1007/s10658-018-1552-2

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  • DOI: https://doi.org/10.1007/s10658-018-1552-2

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