Baculoviruses are known to regulate many insect populations in nature. Their host-specificity is very high, usually restricted to a single or a few closely related insect species. They are amongst the safest pesticides, with no or negligible effects on non-target organisms, including beneficial insects, vertebrates and plants. Baculovirus-based pesticides are compatible with integrated pest management strategies and the expansion of their application will significantly reduce the risks associated with the use of synthetic chemical insecticides. Several successful baculovirus-based pest control programs have taken place in Latin American countries. Sustainable agriculture (a trend promoted by state authorities in most Latin American countries) will benefit from the wider use of registered viral pesticides and new viral products that are in the process of registration and others in the applied research pipeline. The success of baculovirus-based control programs depends upon collaborative efforts among government and research institutions, growers associations, and private companies, which realize the importance of using strategies that protect human health and the environment at large. Initiatives to develop new regulations that promote the use of this type of ecological alternatives tailored to different local conditions and farming systems are underway.

BackgroundEpinotia aporema (Lepidoptera: Tortricidae) is an important pest of legume crops in South America. Epinotia aporema granulovirus (EpapGV) is a baculovirus that causes a polyorganotropic infection in the host larva. Its high pathogenicity and host specificity make EpapGV an excellent candidate to be used as a biological control agent.ResultsThe genome of Epinotia aporema granulovirus (EpapGV) was sequenced and analyzed. Its circular double-stranded DNA genome is 119,082 bp in length and codes for 133 putative genes. It contains the 31 baculovirus core genes and a set of 19 genes that are GV exclusive. Seventeen ORFs were unique to EpapGV in comparison with other baculoviruses. Of these, 16 found no homologues in GenBank, and one encoded a thymidylate kinase. Analysis of nucleotide sequence repeats revealed the presence of 16 homologous regions (hrs) interspersed throughout the genome. Each hr was characterized by the presence of 1 to 3 clustered imperfect palindromes which are similar to previously described palindromes of tortricid-specific GVs. Also, one of the hrs (hr4) has flanking sequences suggestive of a putative non-hr ori. Interestingly, two more complex hrs were found in opposite loci, dividing the circular dsDNA genome in two halves. Gene synteny maps showed the great colinearity of sequenced GVs, being EpapGV the most dissimilar as it has a 20 kb-long gene block inversion. Phylogenetic study performed with 31 core genes of 58 baculoviral genomes suggests that EpapGV is the baculovirus isolate closest to the putative common ancestor of tortricid specific betabaculoviruses.ConclusionsThis study, along with previous characterization of EpapGV infection, is useful for the better understanding of the pathology caused by this virus and its potential utilization as a bioinsecticide.

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae) is among the most voracious global pests of agriculture. Adults of this species were identified recently in northern Argentina by dissection of male genitalia. In this work, a rapid and simple molecular tool was designed to distinguish H. armigera from the morphologically similar indigenous bollworms Helicoverpa zea (Boddie) and Helicoverpa gelotopoeon (Dyar), regardless of the life stage. Amplification of partial COI gene with a new primer pair, and subsequent digestion with endonuclease HinfI, yielded different RFLP profiles for the three main Helicoverpa pests currently present in South America. The method was validated in Helicoverpa specimens collected across Argentina, whose identity was further corroborated by COI sequencing and phylogenetic analysis. The data reported here constitute the first molecular confirmation of this pest in the country. The survey revealed the occurrence of H. armigera in northern and central Argentina, including the main soybean- and maize-producing area.

A granulovirus (GV) isolated from Epinotia aporema (Lepidoptera: Tortricidae)-a major soybean pest-was studied in terms of its main morphological, biochemical, and biological properties. The ovoidal occlusion bodies were 466 by 296 nm in size, and their most prominent protein had an apparent molecular mass of 29 kDa. Its amino-terminal sequence was remarkably homologous to that of the granulins of other GVs. The DNA genome size was estimated to be 120 kbp. The high specificity and pathogenicity of this newly described granulovirus (EpapGV) indicate that it is indeed a good candidate for the biological control of this pest.The bean shoot borer, Epinotia aporema (Wals.), is a serious pest of soybean and other legume crops in South America. In Argentina, damages caused by this tortricid decrease soybean yields up to 40%, depending on population level and environmental conditions (18). Currently, broad-spectrum chemical insecticides are used to control this caterpillar, delaying or suppressing field colonization by natural enemies. This scenario compromises the implementation of a proper integrated pest management scheme for soybean crops, in areas where the incidence of E. aporema reaches high population levels (25).Among pathogens, a granulovirus (GV) was detected as the main mortality factor (6, 26). In general, the potential of baculoviruses for pest control has been well documented and they have proven to be effective against many pests (9,15,24). In this regard, preliminary experiments showed that E. aporema GV (EpapGV) could be a safe alternative for inclusion in soybean pest management (25). However, former studies were directed towards evaluating its virulence, and none of the previously cited reports addressed the characterization of the virus. As an initial point in our research, we identified and characterized a GV isolate from E. aporema, indigenous to the main soybean area of Argentina.Virus isolation and multiplication. A colony of E. aporema caterpillars reared on artificial medium (12) was established in our laboratory. EpapGV was isolated from a single E. aporema larva collected in Oliveros (Santa Fe, Argentina). Viral amplification was carried out, allowing fourth-instar larvae to feed on formalin-free artificial diet, superficially contaminated with 4,000 granules per mm 2 . Granules were purified from homogenized larvae by two cycles of centrifugation on continuous 40 to 65% (wt/wt) sucrose gradients at 100,000 ϫ g for 1 h. Virions were released by hydrolyzing granules in 0.1 M Na 2 CO 3 -0.17 M NaCl-0.01 M EDTA (pH 10.5) at 37°C for 30 min. Following the dissolution of the granules, the suspension was neutralized by addition of 1 M HCl, and undissolved material was removed by low-speed centrifugation.Electron microscopy. Tissues were dissected from infected and control larvae, fixed in 3% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.3)-0.25 M sucrose for 3 h, postfixed in 1% osmium tetroxide for 1 h, dehydrated through an ethanolpropylene oxide series, and embedded in Epon-Araldite resin. Ul...

Anthonomus grandis Boheman is a key pest in cotton crops in the New World. Its larval stage develops within the flower bud using it as food and as protection against its predators. This behavior limits the effectiveness of its control using conventional insecticide applications and biocontrol techniques. In spite of its importance, little is known about its genome sequence and, more important, its specific expression in key organs like the midgut. Total mRNA isolated from larval midguts was used for pyrosequencing. Sequence reads were assembled and annotated to generate a unigene data set. In total, 400,000 reads from A. grandis midgut with an average length of 237 bp were assembled and combined into 20,915 contigs. The assembled reads fell into 6,621 genes models. BlastX search using the NCBI-NR database showed that 3,006 unigenes had significant matches to known sequences. Gene Ontology (GO) mapping analysis evidenced that A. grandis is able to transcripts coding for proteins involved in catalytic processing of macromolecules that allows its adaptation to very different feeding source scenarios. Furthermore, transcripts encoding for proteins involved in detoxification mechanisms such as p450 genes, glutathione-S-transferase , and carboxylesterases are also expressed. This is the first report of a transcriptomic study in A. grandis and the largest set of sequence data reported for this species. These data are valuable resources to expand the knowledge of this insect group and could be used in the design of new control strategies based in molecular information.