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Disruption of prefoldin-2 protein synthesis in root-knot nematodes via host-mediated gene silencing efficiently reduces nematode numbers and thus protects plants

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Abstract

Main conclusion

The aim of this study is to demonstrate the feasibility of down-regulating endogeneous prefoldin-2 root-knot nematode transcripts by expressing dsRNA with sequence identity to the nematode gene in tobacco roots under the influence of strong Arabidopsis ubiquitin (UBQ1) promoter.

Root-knot nematodes (RKNs) are sedentary endoparasites infecting a wide range of plant species. They parasitise the root system, thereby disrupting water and nutrient uptake and causing major reductions in crop yields. The most reliable means of controlling RKNs is via the use of soil fumigants such as methyl bromide. With the emergence of RNA interference (RNAi) technology, which permits host-mediated nematode gene silencing, a new strategy to control plant pathogens has become available. In the present study, we investigated host-induced RNAi gene silencing of prefoldin-2 in transgenic Nicotiana benthamiana. Reductions in prefoldin-2 mRNA transcript levels were observed when nematodes were soaked in a dsRNA solution in vitro. Furthermore, nematode reproduction was suppressed in RNAi transgenic lines, as evident by reductions in the numbers of root knots (by 34–60 % in independent RNAi lines) and egg masses (by 33–58 %). Endogenous expression of prefoldin-2, analysed via real-time polymerase chain reaction and Western blotting, revealed that the gene was strongly expressed in the pre-parasitic J2 stage. Our observations demonstrate the relevance and potential importance of targeting the prefoldin gene during the nematode life cycle. The work also suggests that further improvements in silencing efficiency in economically important crops can be accomplished using RNAi directed against plant-parasitic nematodes.

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Abbreviations

dsRNA:

Double-strand RNA

GUS:

β-Glucuronidase

J2:

Juvenile 2

PFD:

Prefoldin

RKN:

Root-knot nematode

RNAi:

RNA interference

RT-PCR:

Reverse transcription-polymerase chain reaction

UBQ:

Ubiquitin

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Acknowledgments

This project was supported by a grant from the National Academy of Agricultural Science (PJ008541), RDA, Korea. This study was supported by the 2014 Postdoctoral Fellowship Program (to H. Y. C. and I. C. C.) of the National Academy of Agricultural Science, RDA, Korea.

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Authors and Affiliations

  1. National Academy of Agricultural Science, Suwon, 441-857, Korea

    Hemavathi Ajjappala, Ha Young Chung, Joon-Soo Sim, Inchan Choi & Bum-Soo Hahn

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  1. Hemavathi Ajjappala
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  2. Ha Young Chung
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  3. Joon-Soo Sim
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  4. Inchan Choi
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  5. Bum-Soo Hahn
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Correspondence to Bum-Soo Hahn.

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H. Ajjappala and H. Y. Chung contributed equally to this work.

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Ajjappala, H., Chung, H.Y., Sim, JS. et al. Disruption of prefoldin-2 protein synthesis in root-knot nematodes via host-mediated gene silencing efficiently reduces nematode numbers and thus protects plants. Planta 241, 773–787 (2015). https://doi.org/10.1007/s00425-014-2211-2

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