Development of a Recombination System for the Generation of Occlusion Positive Genetically Modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus

Abstract

Anticarsia gemmatalis is an important pest in legume crops in South America and it has been successfully controlled using Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (polh) was replaced by a bacterial β-galactosidase (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome and a transfer vector allowed the restitution of polh and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene (gfp). This recombinant virus infected larvae normally per os and led to the expression of GFP in cell culture as well as in A. gemmatalis larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.

Effects of intraguild interactions on Anticarsia gemmatalis and Chrysodeixis includens larval fitness and behavior in soybean

“…Despite the diverse climate conditions, important soybean producing areas in USA and South American countries are threatened by lepidopteran caterpillars 3–5 . The velvetbean caterpillar, Anticarsia gemmatalis (Lepidoptera: Erebidae), and the soybean looper, Chrysodeixis includens (Lepidoptera: Noctuidae), are among the major soybean pests, which have been causing economic damage from South to North America 6–8 …”

“…[3][4][5] The velvetbean caterpillar, Anticarsia gemmatalis (Lepidoptera: Erebidae), and the soybean looper, Chrysodeixis includens (Lepidoptera: Noctuidae), are among the major soybean pests, which have been causing economic damage from South to North America. [6][7][8] The management of both lepidopteran species can be achieved using different control strategies as part of integrated pest management (IPM) programs, but recently in Brazil, genetically modified soybean expressing a Bacillus thuringiensis (Bt) insecticidal protein has assumed a prominent position for large scale management of lepidopteran insect pests. 9,10 Although this biotechnology is an important tool for the integrated management of A. gemmatalis and C. includens, the use of Bt transgenic crops has been modifying Lepidoptera entomofauna in soybean.…”

Ecofeminismo: Mulheres E Povos Rumo À Uma Cultura Sustentável

“…Segundo estudos conduzidos por Haase et al (2015), a lagarta-da-soja, Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae) é considerada a principal praga desfolhadora da cultura da soja nas Américas, principalmente, nos EUA, México, Colômbia, Venezuela e Brasil. É uma espécie característica de climas tropicais e subtropicais, apresentando de cinco a seis instares larvais, sendo seis o mais comum (SOSA-GOMEZ et al, 2014).…”

Baculovirus Insecticides in Latin America: Historical Overview, Current Status and Future Perspectives

“…Substantial progress has been made for the AgMNPV— A. gemmatalis system, starting with the development of recombinant occlusion-negative baculovirus expressing reporter genes [ 224 , 225 ] and the knockout of the egt gene [ 226 ]. More recently, a highly efficient system to produce AgMNPV recombinants was generated, which will be applied to test alternative genetic modifications aimed at improving biopesticidal parameters [ 227 ]. Additionally, an occlusion-negative AgMNPV was occluded in transgenic cell lines expressing AgMNPV polyhedrin [ 228 ].…”