Open Access
Sequence comparison between three geographically distinct Spodoptera frugiperda multiple nucleopolyhedrovirus isolates: detecting positively selected genes
(Elsevier, 2011-01-14) Simón de Goñi, Oihane; Palma Dovis, Leopoldo; Beperet Arive, Inés; Muñoz Labiano, Delia; López Ferber, Miguel; Caballero Murillo, Primitivo; Williams, Trevor; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
The complete genomic sequence of a Nicaraguan plaque purified Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV) genotype SfMNPV-B was determined and compared to previously sequenced isolates from United States (SfMNPV-3AP2) and Brazil (SfMNPV-19). The genome of SfMNPV-B (132,954 bp) was 1623 bp and 389 bp larger than that of SfMNPV-3AP2 and SfMNPV-19, respectively. Genome size differences were mainly due to a deletion located in the SfMNPV-3AP2 egt region and small deletions and point mutations in SfMNPV-19. Nucleotide sequences were strongly conserved (99.35% identity) and a high degree of predicted amino acid sequence identity was observed. A total of 145 open reading frames (ORFs) were identified in SfMNPV-B, two of them (sf39a and sf110a) had not been previously identified in the SfMNPV-3AP2 and SfMNPV-19 genomes and one (sf57a) was absent in both these genomes. In addition, sf6 was not previously identified in the SfMNPV-19 genome. In contrast, SfMNPV-B and SfMNPV-19 both lacked sf129 that had been reported in SfMNPV-3AP2. In an effort to identify genes potentially involved in virulence or in determining population adaptations, selection pressure analysis was performed. Three ORFs were identified undergoing positive selection: sf49 (pif-3), sf57 (odv-e66b) and sf122 (unknown function). Strong selection for ODV envelope protein genes indicates that the initial infection process in the insect midgut is one critical point at which adaptation acts during the transmission of these viruses in geographically distant populations. The function of ORF sf122 is being examined.
Open Access
Mixtures of complete and pif1- and pif2-deficient genotypes are required for increased potency of an insect nucleopolyhedrovirus
(American Society for Microbiology, 2009) Clavijo Palacios, Gabriel; Williams, Trevor; Simón de Goñi, Oihane; Muñoz Labiano, Delia; Cerutti, Martine; López Ferber, Miguel; Caballero Murillo, Primitivo; Nekazaritza Ekoizpena; Producción Agraria; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
The insecticidal potency of a nucleopolyhedrovirus population (SfNIC) that infects Spodoptera frugiperda (Lepidoptera) is greater than the potency of any of the component genotypes alone. Occlusion bodies (OBs) produced in mixed infections comprising the complete genotype and a deletion genotype are as pathogenic as the natural population of genotypes from the field. To test whether this increased potency was due to the deletion or to some other characteristic of the deletion variant genome, we used the SfNIC-B genome to construct a recombinant virus (SfNIC-BΔ16K) with the same 16.4-kb deletion as that observed in SfNIC-C and another recombinant (SfNIC-BΔpifs) with a deletion encompassing two adjacent genes (pif1 and pif2) that are essential for transmission per os. Mixtures comprising SfNIC-B and SfNIC-B 16K in OB ratios that varied between 10:90 and 90:10 were injected into insects, and the progeny OBs were fed to larvae in an insecticidal potency assay. A densitometric analysis of PCR products indicated that SfNIC-B was generally more abundant than expected in mixtures based on the proportions of OBs used to produce the inocula. Mixtures derived from OB ratios of 10, 25, or 50% of SfNIC-BΔ16K and the corresponding SfNIC-B proportions showed a significant increase in potency compared to SfNIC-B alone. The results of potency assays with mixtures comprising various proportions of SfNIC-B plus SfNIC-BΔpifs were almost identical to the results observed with SfNICB 16K, indicating that deletion of the pif gene region was responsible for the increased potency observed in mixtures of SfNIC-B and each deletion recombinant virus. Subsequently, mixtures produced from OB ratios involving 10 or 90% of SfNIC-BΔ16K with the corresponding proportions of SfNIC-B were subjected to four rounds of per os transmission in larvae. The composition of each experimental mixture rapidly converged to a common equilibrium with a genotypic composition of ~85% SfNIC-B plus 15% SfNIC-BΔ16K. Nearly identical results were observed in peroral-passage experiments involving mixtures of SfNIC-B plus SfNICBΔpifs. We conclude that (i) the deletion of the pif1 and pif2 region is necessary and sufficient to explain the increased potency observed in mixtures of complete and deletion genotypes and (ii) viral populations with decreased ratios of pif1- and pif2-deficient genotypes in the virus population increase the potency of genotypic mixtures and are likely to positively influence the transmission of this pathogen.
Open Access
Defective or effective?: mutualistic interactions between virus genotypes
(Royal Society, 2003-11-07) López Ferber, Miguel; Simón de Goñi, Oihane; Williams, Trevor; Caballero Murillo, Primitivo; Producción Agraria; Nekazaritza Ekoizpena
Defective viruses lack genes essential for survival but they can co-infect with complete virus genotypes and use gene products from the complete genotype for their replication and transmission. As such, they are detrimental to the fitness of complete genotypes. Here, we describe a mutualistic interaction between genotypes of an insect baculovirus (nucleopolyhedrovirus of Spodoptera frugiperda (Lepidoptera)) that increases the pathogenicity of the viral population. Mixtures of a complete genotype able to be transmitted orally and a deletion mutant unable to be transmitted orally resulted in a phenotype of increased pathogenicity. Because the infectiousness of mixed genotype infections was greater than that of single genotype infections, we predict that the transmissibility of mixed genotype occlusion bodies will be greater than that of any of their single genotype components. Such interactions will be subject to frequency-dependent selection and will influence the impact of these viruses on insect population dynamics and their efficacy as biological insecticides.
Open Access
Physical and partial genetic map of Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV) genome
(Springer, 2005) Simón de Goñi, Oihane; Chevenet, François; Williams, Trevor; Caballero Murillo, Primitivo; López Ferber, Miguel; Producción Agraria; Nekazaritza Ekoizpena
A Nicaraguan isolate of Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV) is undergoing field trials for control of this pest in the Americas. This isolate is composed of multiple genotypes, some of which are deletion mutants. Identification of the genetic changes in deleted genotypes cannot be accomplished without the construction of a detailed physical map. In the present study, combinations of restriction endonuclease analysis and Southern blot analysis was performed. This map was refined by sequencing the termini of cloned restriction fragments. The SfMNPV genome was estimated to be 129.3 kb, 8 kb larger than the previously characterized Sf-2 variant from the United States, due to a deletion between 14.8 and 21.0 m.u. in the physical map described in this study. A total of 27.92 kb were sequenced, which represented 21.5% of the whole genome and included 38 ORFs. Comparison with other sequenced baculoviruses revealed that SfMNPV displayed the highest sequence identity (66%) and gene arrangement (78%) with Spodoptera exigua MNPV, sharing 36 putative ORFs. In addition, the genome organization was similar to that of SeMNPV, with minor differences. Phylogenetic analysis confirmed the close relatedness between SeMNPV and SfMNPV, suggesting they evolved from a common ancestor.
Open Access
Occlusion body pathogenicity, virulence and productivity traits vary with transmission strategy in a nucleopolyhedrovirus
(Elservier, 2011-10-25) Cabodevilla de Andrés, Oihana; Ibáñez Elosua, Itxaso; Simón de Goñi, Oihane; Murillo Pérez, Rosa; Caballero Murillo, Primitivo; Williams, Trevor; Producción Agraria; Nekazaritza Ekoizpena; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
The prevalence of sublethal infections of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) was quantified in natural populations of S. exigua in Almería, Spain, during 2006 and 2007. Of 1045 adults collected, 167 (16.1%) proved positive for viral polyhedrin gene transcripts by RT-PCR. The prevalence of covert infection varied significantly according to sex and sample date. Of 1660 progeny of field-collected insects, lethal disease was observed in 10¿33% of offspring of transcript-positive females and 9¿49% of offspring of transcript-negative females. Isolates associated with vertically transmitted infections were characterized by restriction endonuclease analysis using BglII or EcoRV and compared with isolates originating from greenhouse soil-substrate believed to be horizontally transmitted. Insects from a sublethally infected Almerian colony were between 2.3-fold and 4.6-fold more susceptible to infection than healthy insects from a Swiss colony, depending on isolate. Horizontally transmitted isolates were significantly more pathogenic than vertically transmitted isolates in insects from both colonies. Mean speed of kill in second instars (Swiss colony) varied between isolates by >20 h, whereas mean occlusion body (OB) production in fourth instars (Swiss colony) varied by 3.8-fold among isolates. Intriguingly, all three horizontally transmitted isolates were very similar in speed of kill and OB production, whereas all three vertically transmitted isolates differed significantly from one another in both variables, and also differed significantly from the group of horizontally transmitted isolates in speed of kill (one isolate) or both variables (two isolates). We conclude that key pathogenicity and virulence traits of SeMNPV isolates vary according to their principal transmission strategy.
Open Access
A native variant of Chrysodeixis chalcites nucleopolyhedrovirus: the basis for a promising bioinsecticide for control of C. chalcites in Canary Islands' banana crops
(Elsevier, 2013-08-13) Bernal Rodríguez, Alexandra; Williams, Trevor; Hernández Suárez, Estrella; Carnero, Aurelio; Caballero Murillo, Primitivo; Simón de Goñi, Oihane; Producción Agraria; Nekazaritza Ekoizpena; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Chrysodeixis chalcites (Lepidoptera: Noctuidae) larvae cause up to 30% production loss in banana crops in the Canary Islands. Larvae of this species are susceptible to a nucleopolyhedrovirus (ChchNPV). This study aimed at evaluating the genetic diversity and bioinsecticidal activity of ChchNPV isolates collected from C. chalcites larvae in the Canary Islands. From a total 97 isolates collected in different banana greenhouses, restriction endonuclease analysis identified five genetic variants that differed slightly from ChchNPV isolates from Netherlands (ChchSNPV-NL) and Almería, Spain (ChchNPV-SP1). Physical maps revealed minimal differences at the genome level, mostly due to variation in the position/existence of restriction sites. ChchSNPV-TF1 was the most prevalent variant, representing 78% of isolates examined, and was isolated at all Canary Island sampling sites. This isolate was the most pathogenic isolate against C. chalcites second instars in terms of concentration-mortality metrics, compared to homologous variants or two heterologous viruses Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Anagrapha falcifera multiple nucleopolyhedrovirus (AnfaMNPV). ChchSNPV-TF1 was also one of the fastest killing variants although no differences were observed in occlusion body production among the different variants in second instars. We conclude that ChchSNPV-TF1 merits further evaluation as the basis for a biological insecticide for control of C. chalcites in banana crops in the Canary Islands.