Cunea Larvae Research Articles

Comparative analysis of two attacin genes from Hyphantria cunea

A full-length clone corresponding to attacin was isolated from a cDNA library made from fat body of immunized Hyphantria cunea larvae. This newly isolated attacin B shows characteristics different from those previously reported for attacin A. The two attacin cDNAs encode precursor proteins of 233 and 248 amino acid residues, respectively. The two attacins show 45.9% identity at the amino acid level, and 35.2% identity at the nucleotide level. Attacins A and B of H. cunea show significant identities with the attacins of Lepidoptera. Attacin B is a typical glycine-rich protein, while attacin A is leucine-rich. Attacin B is expressed from last instar larvae to adult, while attacin A showed stage-specific expression during the prepupal and pupal stages. Attacins A and B are predicted to have different secondary structure in that attacin A has no tendency to form helices but attacin B contains a substantial number of helices. Attacin A is induced at a trace level in infected larvae, while attacin B is strongly induced against Gram-positive and negative bacteria, fungi, and viruses. The attacin B transcripts were detected in fat body, epidermis and hemocytes after injection with Escherichia coli, Citrobacter freundii, or Candida albicans, but not in the midgut and Malpighian tubule. Recombinant attacin A showed no antibacterial activity, while recombinant attacin B showed strong antibacterial activity in proportion to the amount of the protein injected.

Two Clip Domain Family of Serine Proteases and a Serine Protease Homolog from the Fall Webworm, Hyphantria cunea; cDNA Cloning and Characterization

ABSTRACTTwo types of serine proteases and a serine protease homologue cDNAs were isolated from Hyphantria cunea larvae induced immune response due to an injection of a microorganism through RT‐PCR and cDNA library screening, and their characteristics were examined. The isolated cDNAs are composed 2.1 kb, 2.2 kb, and 2.5 kb nucleotide each, which encoded 388, 390, 580 amino acid residues, and were designated as HcPE‐1, HcPE‐2 and HcPE‐3, respectively. They were revealed as serine proteases or a serine protease homologue with the clip domain through a database search. The deduced amino acid sequence comparison showed high homology of 72‐78% among them. Six Cys residues of the N‐terminal clip domain forming the disulfide bond, Cys residues of the catalytic domain, and Cys residues forming inter‐bridge between clip domain and catalytic domain were also well preserved. Three amino acid residues, His, Asp, and Ser, within the active site were perfectly conserved in HcPE‐2 and HcPE‐3, however, His was replaced with Gln178 in HcPE‐1. The Arg residues (HcPE‐1, Arg132; HcPE‐2, Arg134; HcPE‐3, Arg325) known as the activation sites by proteolytic cleavage were preserved well in all three types of protein. In case of HcPE‐3, three continuous clip‐like domains existed in the N terminal. As the result of phylogenetic analysis, three clip domain family of protein from H. cunea make groups with arthropod proclotting enzyme precursor. Northern blot analysis showed all three genes were induced through an injection of Escherichia coli, but expression patterns were varied.

Antifeeding activity of isoquinoline alkaloids identified in Coptis japonica roots against Hyphantria cunea (Lepidoptera: Arctiidae) and Agelastica coerulea (Coleoptera: Galerucinae).

The anti-feeding activity of 3 isoquinoline alkaloids identified from roots of Coptis japonica Makino toward 4th-instar larvae of Hyphantria cunea Drury and adults of Agelastica coerulea Baly was examined using the leaf-dipping bioassay. The biologically active constituents of the Coptis roots were characterized as the isoquinoline alkaloids berberine, palmatine and coptisine by spectroscopic analysis. In a test with H. cunea larvae, the anti-feeding activity was much more pronounced in an application of a mixture of palmatine iodide and berberine chloride (1:1, wt:wt) at 250 ppm (82.3%) and 500 ppm (100%), compared with palmatine iodide (76.0%) and berberine chloride (75.4%) alone at 500 ppm. These results indicate a synergistic effect. With A. courulea adults, berberine chloride showed 57.5 and 91.1% anti-feeding activity at 125 and 250 ppm, respectively; whereas, weak activity was obtained in application of 500 ppm of palmatine iodide (41.4%) and coptisine chloride (52.4%) alone. The Coptis root-derived compounds merit further study as potential insect-control agents.

Susceptibility of Hyphantria cunea infected with the Diacrisia granulosis virus to its homologous baculoviruses

The interactions between the Diacrisia virginica granulosis virus (DGV) and the Hyphantria cunea baculovirus isolates were determined, utilizing defined differences between the time-mortality responses of these viruses fed to H. cunea larvae. The DGV, having a prolonged incubation period, when given an advantage in time or in the number of capsules, was able to prevent the expression of the more lethal H. cunea granulosis virus (HcGV) isolate. The time-mortality response of test larvae simultaneously fed equivalent dosages of HcGV and DGV was intermediate to that achieved with HcGV alone or DGV alone. Larvae infected with the DGV isolate were still susceptible to double infection by the nucleopolyhedrosis virus. The time-mortality response demonstrated that the development of nucleopolyhedrosis was only partly inhibited by preinfecting the test larvae with the DGV isolate.

Susceptibility of Hyphantria cunea to a granulosis virus isolated from Diacrisia virginica

The dosage-mortality response of Hyphantria cunea larvae to a granulosis virus isolated from Diacrisia virginica was studied. Serial decimal dilutions of the D. virginica granulosis virus were fed to early second-instar H. cunea larvae. the LD 50 for this virus (7.9 × 10 5 capsules/larva) was significantly greater than the LD 50 calculated for the H. cunea granulosis virus (7.06 × 10 4 capsules/larva) against the same instar of H. cunea. Time mortality studies demonstrated that the LT 50 values for the D. virginica granulosis were four to six times greater than for comparable dosages of H. cunea granulosis virus. Based on the mortality response of assayed H. cunea, it is possible to distinguish between the D. virginica granulosis virus and the H. cunea granulosis virus.

PREDATION BY INSECTS AND SPIDERS INHABITING COLONIAL WEBS OF HYPHANTRIA CUNEA

AbstractThe number of predators inhabiting nests of Hyphantria cunea Drury was recorded annually for 13 years in four areas in New Brunswick and two areas on the coast of Nova Scotia. The most common groups were the pentatomids and spiders, which sometimes reproduced within the nests, but the mean number per nest was low in relation to the number of H. cunea larvae in the colonies. The rate of predation on fifth-instar larvae was low. Small or timid predators appeared to prey largely on moribund larvae or small saprophagans during the principal defoliating instars of H. cunea.No relationship could be detected between the number of larvae reaching the fifth instar and the number of predators in the colony; nor could any functional or numerical response of the predators to either the initial number of larvae per colony or the population density of colonies be found. It is concluded that the influence of the nest-inhabiting predators is small and relatively stable, and may be treated as a constant in the development of models to explain the population dynamics of H. cunea.H. cunea is a pest in parts of Europe and Asia, where it has been accidentally introduced from North America. The introduction to other continents of the North American predator, Podisus maculiventiis (Say), is discussed briefly.