Pseudoplusia includens single nucleopolyhedrovirus: Genetic diversity,phylogeny and hypervariability of the pif-2 gene

The soybean looper (Pseudoplusia includens Walker, 1857) has become a major pest of soybean crops in Brazil. In order to determine the genetic diversity and phylogeny of variants of Pseudoplusia includens single nucleopolyhedrovirus (PsinSNPV-IA to -IG), partial sequences of the genes lef-8, lef-9, pif-2, phr and polh were obtained following degenerate PCR and phylogenetic trees constructed using maximum parsimony and Bayesian methods. The aligned sequences showed polymorphisms among the isolates, where the pif-2 gene was by far the most variable and is predicted to be under positive selection. Furthermore, some of the pif-2 DNA sequence mutations are predicted to result in significant amino acid substitutions, possibly leading to changes in oral infectivity of this baculovirus. Cladistic analysis revealed two closely related monophyletic groups, one containing PsinNPV isolates IB, IC and ID and another containing isolates IA, IE, IF and IG. The phylogeny of PsinSNPV in relation to 56 other baculoviruses was also determined from the concatenated partial LEF-8, LEF-9, PIF-2 and POLH/GRAN deduced amino acid sequences, using maximum-parsimony and Bayesian methods. This analysis clearly places PsinSNPV with the Group II Alphabaculovirus, where PsinSNPV is most closely related to Chrysodeixis chalcites NPV and Trichoplusia ni SNPV.     

Insect resistance in recombinant inbred soybean lines derived from non-resistant parents

Larval performance of Helicoverpa zea (Boddie) (corn earworm) (Lepidoptera: Noctuidae) was examined on 240 recombinant inbred (RI) soybean, Glycine max (L.) Merrill, lines. These homozygous RI were derived from an intraspecific cross of genetically distant, non-resistant, parents, Minsoy from China and Noir 1 from Hungary. Based upon a genetic map of more than 500 molecular markers, each RI line presented a unique genotype composed of a mixture of different parental alleles. The RI lines exhibited transgressive segregation with respect to their defensive effects on H. zea, such that the range of RI phenotypes far exceeded that of the parents. Similar effects were observed on the soybean looper, Pseudoplusia includens (Walker) (Lepidoptera: Noctuidae). We identified several independent quantitative trait loci (QTLs) linked to molecular markers that were associated with H. zea larval development parameters. Two QTLs affected several different traits including larval weight and developmental rate; other QTLs affected only a single trait each, i.e., larval weight, pupal weight, developmental rate, nutritional efficiency or survival. The results demonstrate that the increased range of defensive effects among the segregant RI lines is due to recombination among several parental genes that together quantitatively control plant defensive traits.Several alternative responses by herbivores have been proposed relative to plant hybrid swarms, hybrid avoidance due to higher hybrid resistance than either parent, hybrid preference due to lower resistance than either parent, hybrid equivalency to one or the other parent, or hybrid intermediacy. Within this RI population, we observed all of the proposed responses by H. zea, as might be expected when defensive traits are controlled by several genes.     

Influence of induced resistance in soybean on the development and nutrition of the soybean looper and the Mexican bean beetle

Standard gravimetric technique was used to assess the effect of induced resistance in soybean on the consumption and utilization of food by the Mexican bean beetle (MBB), Epilachna varivestis, and the soybean looper (SBL), Pseudoplusia includens. Induced resistance had significant retarding effects on the development and growth of both SBL and MBB. SBL fed leaves of previously injured soybean plants had a 8.5% longer developmental time through the entire larval stage and a 10.4% lower pupal weight than larvae fed leaves from uninjured plants. MBB fed on leaves from previously injured plants through the first three instars had a 9.1% longer developmental time and a 16.5% lower final weight than those fed on leaves from unijured plants. Induced resistance showed no significant effect on total food consumed by either SBL or MBB. Nutritional indices including relative consumption rate (RCR), relative growth rate (RGR), efficiency of conversion of ingested food (ECI), approximate digestibility (AD), and efficiency of conversion of digested food (ECD) were calculated on dry weight measurements. In general, induced resistance had greater impacts on the performance of MBB than on that of SBL.

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Résumé Une technique gravimétrique uniforme a été utilisée pour mesurer l'effet de la résistance induite du soja et la consommation et l'utilisation d'aliments par E. varivestis (Coléop.: Coccinellidae) et P. includens (Lepid.: Noctuidae). La résistance induite a significativement retardé la croissance et le développement des 2 insectes. La consommation de feuilles déjà abîmées a prolongé de 8.5% la durée du développement de P. includens et a réduit de 10.4% le poids des chrysalides. La consommation de feuilles de soja abîmées pendant les 3 premiers stades d'E. varivestis a prolongé le développement de 9.1% et a réduit de 16.5% le poids des nymphes. La résistance induite n'a pas provoqué une modification significative de la quantité de feuilles de soja abîmées chez les 2 insectes. Les indices de nutrition, comprenant le taux de consommation relative (RCR), le taux de croissance relative (RGR), l'efficacité de transformation de l'aliment ingéré (ECI), la digestibilité approchée (AD) et l'efficacité de conversion de l'aliment digéré (ECD), ont été calculés à partir des poids secs. En général, la résistance induite a eu des effets plus nets sur les performances d'E. varivestis que de P. includens.

     

Population Development of Meloidogyne incognita on Soybean Defoliated by Pseudoplusia includens

Greenhouse studies examined population densities of Meloidogyne incognita race 4 on soybean (Glycine max ''Davis'') defoliated by larvae of soybean looper (Pseudoplusia indudens (Walker)). Plants were defoliated over a 2-week period beginning 5 weeks after seedlings were transplanted. Four groups of plants were infested with nematodes (5,000 eggs/pot) at 2-week intervals to allow harvesting of plants at 0, 2, 4, and 6 weeks postdefoliation (WPD). Plants in each group were harvested 4 weeks after nematode infestation. Root and nodule weights of defoliated plants were suppressed at 0 WPD, but differences were not detectable at 2, 4, and 6 WPD. Population densities of M. incognita were similar on defoliated and control plants at 0 WPD but were greater on defoliated plants at 4 and 6 WPD. Percentage hatching of eggs produced on the latter plants also was higher. Effects of insect-induced defoliation on development of M. incognita remained detectable even after soybean plant growth apparently returned to normal.     

Biological and molecular characterization of a multicapsid nucleopolyhedrovirus from Thysanoplusia orichalcea (L.) (Lepidoptera: Noctuidae)

A multicapsid nucleopolyhedrovirus (ThorMNPV) that was co-isolated with a single nucleocapid ThorSNPV from mixed infected larvae of Thysanoplusia orichalcea L. (Lepidoptea: Noctuidae) is characterized. Scanning electron microscopy of ThorMNPV showed a dodecahedral-shaped occlusion body (OB). The occluded virions contained one to as many as eight nucleocapsids/virion. Virion band profiles in gradient centrifugation were consistent in at least 10 rounds of centrifugation from different virion sample preparations. The ThorMNPV had high virulence to third instar Trichoplusia ni and Pseudoplusia includens with LD50 values of 17 and 242OBs per larva, respectively. However, ThorMNPV did not cause mortality in Spodoptera exigua, Spodoptera frugiperda, Spodoptera eridania, Anticarsia gemmatalis, and Helicoverpa zea. ThorMNPV replicates in cells of various tissues such as the fat body and tracheal epithelium cells. T. ni High 5 cells were permissive to ThorMNPV in terms of infection and viral DNA transfection, but SF-21 was less permissive and the infection process was slower. Production of OBs by ThorMNPV in the nuclei of SF-21 was not well pronounced. The genome size of ThorMNPV was estimated to be 136 kb. The polyhedrin gene open reading frame (ORF) was cloned and completely sequenced. The promoter sequence is identical to that of Autographa californica MNPV. Phylogenetic analyses using partial sequences of the polh, lef-8, and lef-9 revealed that ThorMNPV is a member of the Group I NPVs and is related but distinct from the AcMNPV/Rachiplusia ou NPV/Bombyx mori NPV cluster.     

The action of acephate in Pseudoplusia includens (Lepidoptera: Noctuidae) and Pristhesancus papuensis (Hemiptera: Reduviidae)

The toxicity, toxodynamics, and toxokinetics of acephate were investigated in the soybean looper, Pseudoplusia includens, and a predatory reduviid, Pristhesancus papuensis. Acephate was nine-fold more toxic to the predator than the phytophagous soybean looper at the LD₅₀. Both acephate and its presumed activation product, methamidophos, were found to be inhibitors of acetylcholinesterase (AcChE) in vitro. Acetylcholinesterase from P. includens was more susceptible to inhibition by both chemicals than was P. papuensis AcChE. The overall rate of penetration, metabolism, and excretion was more rapid in P. includens than in the predator. The activation of acephate to methamidophos was four times greater in P. papuensis than in P. includens after a 24-h exposure period. No labeled CO₂ was detected from P. papuensis, while over 5% of the applied dose was trapped as CO₂ from P. includens. The activation and accumulation steps then, are apparently the major reasons for the greater toxicity of acephate to P. papuensis compared to P. includens.

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Résumé L'action de l'acéphate (orthène) a été déterminée sur une lignée de Pseudoplusia includens et un prédateur éventuel, Pristhesancus papuensis. Appliqué localement, l'acéphate a été très toxique pour les 2 espèces, bien que le prédateur ait été 9 fois plus sensible que la noctuelle à la DL50.Tant l'acéphate qu'un composé similaire, le méthamidophos, ont inhibé l'acétylcholinestérase (AcChE) in vitro. Le pouvoir inhibiteur du méthamidophos était, cependant, 1 000 fois plus puissant que celui de l'acéphate sur cette enzyme. L'enzyme de P. includens a été plus susceptible d'inhibition par ces 2 substances que l'AcChE de P. papuensis.La pénétration de la radioactivité à la suite d'un traitement au ¹⁴C-acéphate a été rapide chez P. includens, chez lequel au bout de 48 h il y avait eu accumulation interne de 43% de la dose appliquée. La pénétration de l'acéphate chez P. papuensis a été moins rapide, n'atteignant que 21% au bout de 48 h. Le taux de CO₂ marqué dégagé 24 h après le traitement a été déterminé en utilisant un collecteur de CO₂ respiratoire et un piège. Aucun CO₂ marqué n'a été décelé à partir du prédateur, tandis que plus de 5% de la dose appliquée a été piégée sous forme de CO₂ chez la noctuelle. P. papuensis a activé in vivo 4 fois la proportion d'acéphate appliqué par rapport au méthamidophos que ne l'a fait P. includens; cependant aucun d'eux n'a été éliminé dans les fécès. D'autre part, chez P. includens, le méthamidophos et l'acéphate non métabolisé ont été perdus dans le bocal d'élevage, et seulement la moitié du taux de radioactivité interne était présent au bout de 24 h. Les étapes de l'activation et de l'accumulation ont été les principales raisons de la plus grande toxicité de l'acéphate pour le prédateur.

     

Complete genome sequence of Bacillus thuringiensis BR145, a strain with insecticidal activity against Lepidoptera pests

Bacillus thuringiensis BR145 isolated from a soybean field in Southern Brazil showed toxicity against two important insect pests from soybean crop, Helicoverpa armigera, and Chrysodeixis includens, with LC₅₀ 0.294 µg.cm^-2 and 0.277 µg.cm^-2, respectively. We analyzed the genome of this strain through sequences obtained by Next Generation DNA Sequencing and de novo assembly. The analysis of the genome revealed insecticidal genes cry1Aa, cry1Ab, cry1Ac, cry1Ia, cry2Ab, cyt1, and vip3Aa, suggesting the use of this strain in new strategies of biological control.     

Response of five insect herbivores to multiple allelochemicals under fluctuating temperatures

Analysis of the combined effects of allelochemicals on insect herbivores is useful because there may be adverse additive or even synergistic effects. Analysis of the simultaneous effects of temperature and alleochemicals is also necessary because these factors may interact. We examined the effects of three allelochemicals found in tomato (chlorogenic acid, rutin and tomatine) and thermal regime (21:10 °C and 26:15 °C, representing spring and summer respectively) on five insect herbivores (a Solanaceae specialist, Manduca sexta, and the polyphagous Heliothis virescens, Pseudoplusia includens, Spodoptera frugiperda and Trichoplusia ni). There were allelochemical interactions and thermal regime-allelochemical interactions for all species, and so the patterns were complex. In some cases, paired allelochemicals or the combination of three allelochemicals showed adverse additive effects on insect performance. But that was not always the case, and there were only a few examples of synergism. Negative effects of the allelochemicals were sometimes, but not always, damped by the cooler thermal regime. Comparing the growth rates of the five species in this study with those of a previous study (a total of seven species) revealed five patterns. For two of three pairs of closely-related species, the paired species had distinctly different patterns. For example, for H. virescens, tomatine prevented development and chlorogenic acid slowed growth, whereas for Helicoverpa zea, tomatine just slowed growth and the phenolics had little effect. The specialist Manduca sexta had a pattern that was midway between patterns of the generalists; it was not the most tolerant of the allelochemicals.     

Development of Heterodera glycines on Soybean Damaged by Soybean Looper and Stem Canker

Short-term greenhouse studies with soybean (Glycine max cv. Bragg) were used to examine interactions between the soybean cyst nematode (Heterodera glycines) and two other common pests of soybean, the stem canker fungus (Diaporthe phaseolorum var. caulivora) and the soybean looper (Pseudoplusia includens), a lepidopterous defoliator. Numbers of cyst nematode juveniles in roots and numbers of cysts in soil and roots were reduced on plants with stem cankers. Defoliation by soybean looper larvae had the opposite effect; defoliation levels of 22 and 64% caused stepwise increases in numbers of juveniles and cysts in both roots and soil, whereas numbers of females in roots decreased. In two experiments, stem canker length was reduced 40 and 45% when root systems were colonized by the soybean cyst nematode. The absence of significant interactions among these pests indicates that the effects of soybean cyst nematode, stem canker, and soybean looper on plant growth and each other primarily were additive.