ABSTRACT
In search for local entomopathogenic nematode (EPN) species as a biological control agent of lepidopterous insect pests of corn, a survey for EPN in the major islands in the Philippines was conducted. Seven EPN populations from 279 soil samples were isolated using Ostrinia furnacalis, the key target insect pest of corn in the country, as bait. Analysis of the ITS1-5.8S-ITS2 ribosomal DNA sequence revealed the presence of Steinernema abbasi, Steinernema minutum , Steinernema tami , and Heterorhabditis indica . The pathogenicity of these EPN was tested in Ostrinia furnacalis , Spodoptera litura , and Helicoverpa armigera larvae under laboratory conditions. All the EPN isolates were pathogenic to the lepidopteran species with, H. indica PBCB and S. abbasi MBLB exhibiting the highest virulence (88%-99.33% and 90%-100% mortality, respectively) at 48 hr post infection (HPI) and thus, further studies were done on these two EPN. The highest penetration rate at 48 HPI was observed in H. armigera infected with S. abbasi MBLB (28.15%), while the lowest was in O. furnacalis infected with H. indica PBCB (14.25%). Nonetheless, based on LC 50 at 48 HPI, H. indica PBCB was most virulent to S. litura (8.89 IJ per larva), but not significantly different from O. furnacalis (10.52 IJ per larva). Steinernema abbasi MBLB was most virulent to O. furnacalis (10.98 IJ per larva), but not significantly different to S. litura (17.08 IJ per larva). LT 50 estimates showed that O. furnacalis was significantly the most susceptible to H. indica PBCB (21.90 hr) and S. abbasi (21.18 hr). Our results suggest that H. indica PBCB and S. abbasi MBLB are good candidates as biological control agents against these insect pests of corn. Moreover, O. furnacalis as alternative bait for EPN was discussed. To date, this is the most extensive research on Philippine EPN, comprised of wide sampling coverage, molecular identification and bioefficacy assays.
ABSTRACT
A newly cloned Helicoverpa armigera nucleopolyhedrovirus (HearNPV) from Kenya, HearNPV-NNg1, has a higher insecticidal activity than HearNPV-G4, which also exhibits lower insecticidal activity than HearNPV-C1. In the search for genes and/or nucleotide sequences that might be involved in the observed virulence differences among Helicoverpa spp. NPVs, the entire genome of NNg1 was sequenced and compared with previously sequenced genomes of G4, C1 and Helicoverpa zea single-nucleocapsid NPV (Hz). The NNg1 genome was 132,425 bp in length, with a total of 143 putative open reading frames (ORFs), and shared high levels of overall amino acid and nucleotide sequence identities with G4, C1 and Hz. Three NNg1 ORFs, ORF5, ORF100 and ORF124, which were shared with C1, were absent in G4 and Hz, while NNg1 and C1 were missing a homologue of G4/Hz ORF5. Another three ORFs, ORF60 (bro-b), ORF119 and ORF120, and one direct repeat sequence (dr) were unique to NNg1. Relative to the overall nucleotide sequence identity, lower sequence identities were observed between NNg1 hrs and the homologous hrs in the other three Helicoverpa spp. NPVs, despite containing the same number of hrs located at essentially the same positions on the genomes. Differences were also observed between NNg1 and each of the other three Helicoverpa spp. NPVs in the diversity of bro genes encoded on the genomes. These results indicate several putative genes and nucleotide sequences that may be responsible for the virulence differences observed among Helicoverpa spp., yet the specific genes and/or nucleotide sequences responsible have not been identified.
