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Midgut Genes Knockdown by Oral dsRNA Administration Produces a Lethal Effect on Cotton Boll Weevil

  • Pest Management
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Abstract

The “cotton boll weevil” (Anthonomus grandis Boheman) is a key pest in America whose larval stage develops within the cotton flower bud. During its development, the larva uses the flower bud as food and as a shelter from predators. This behavior limits the effective control through conventional insecticide applications and biocontrol techniques. Increasing genetic information from insects has allowed the development of new control technologies based on the use of RNA interference (RNAi) to design orally delivered double-stranded RNA (dsRNA) strategies. In this study, we evaluated the effect of continuous oral administration of six specific dsRNA in order to identify an effective target gene for RNAi-mediated control of cotton boll weevil. First, six selected A. grandis gene fragments were amplified and cloned to perform in vivo synthesis of the specific dsRNA, and subsequently, larvae and adults were fed with this dsRNA for 2 weeks. Larvae mortality ranged from 40 to 60% depending on the targeted gene sequence. Indeed, α-amylase and cytochrome p450 dsRNAs were the most effective. Oral administration in adults caused smaller but still significant death rates (15–30%). Thus, the results demonstrated RNAi responses depend on life stages and target genes. The dsRNA ingestion was capable of providing knockdown mRNA levels in cotton boll weevil midgut and this effect was significantly higher in the larval stage. In this study, we present a new report of silencing of midgut genes in A. grandis larva induced by continuously feeding with dsRNA. This potential new tool should be further evaluated in cotton boll weevil control strategies.

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Acknowledgments

We thank Julia Sabio y García for assistance with the English edition. Addgene Inc. (Cambridge, MA, USA) provided the plasmid L4440, whereas Caenorhabditis Genetics Center (CGC, Minneapolis, MN, USA), which is funded by the NIH National Center for Research Resources (NCCR), kindly provided the bacterial strain HT115(DE3). Most of the work was supported by a joint venture project entitled “Knowledge generation and development of non-pollutant biotechnologies for the control of cotton weevil” signed between INTA and the governments of Chaco, Formosa, Santa Fe, and Santiago del Estero provinces (Argentina).

Funding

Most of the work was supported by a joint venture project entitled “Knowledge generation and development of non-pollutant biotechnologies for the control of cotton weevil” signed between the governments of Chaco, Formosa, Santa Fe, and Santiago del Estero provinces (Argentina) and the National Institute of Agricultural Technology (INTA).

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Authors and Affiliations

  1. Instituto de Microbiología y Zoología Agrícola (IMyZA), Centro de investigaciones en Ciencias Agronómicas y Veterinarias (CICVyA), Instituto Nacional de Tecnología Agropecuaria (INTA), Buenos Aires, Argentina

    Ricardo Salvador, José M. Niz, Pablo A. Nakaya & Analía Pedarros

  2. Instituto de Agrobiotecnología y Biología Molecular (IABIMO), Instituto Nacional de Tecnología Agropecuaria (INTA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and Lab de Agrobiotecnología DFBMC, Facultad de Ciencias Exactas y Naturales, Univ de Buenos Aires, Buenos Aires, Argentina

    H. Esteban Hopp

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  1. Ricardo Salvador
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  2. José M. Niz
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  3. Pablo A. Nakaya
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  4. Analía Pedarros
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  5. H. Esteban Hopp
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Contributions

Conceptualization: Salvador, Ricardo; Niz, José; Pedarros, Analia; Nakaya, Pablo; and Hopp, Esteban. Investigation: Salvador, Ricardo; Niz, José; Pedarros, Analía; and Nakaya, Pablo. Supervision: Salvador, Ricardo and Hopp, Esteban. Writing original draft: Salvador, Ricardo. Writing review and editing: Salvador, Ricardo and Hopp, Esteban. Funding acquisition: Hopp, Esteban.

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Correspondence to Ricardo Salvador.

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Salvador, R., Niz, J.M., Nakaya, P.A. et al. Midgut Genes Knockdown by Oral dsRNA Administration Produces a Lethal Effect on Cotton Boll Weevil. Neotrop Entomol 50, 121–128 (2021). https://doi.org/10.1007/s13744-020-00819-1

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