Yellow-spotted Longicorn Research Articles

Antiseptic effect of antimicrobial peptide psacotheasin 2 derived from the yellow-spotted longicorn beetle (Psacothea hilaris)

Given the challenges posed by antibiotic resistant microbes and the high mortality rate associated with sepsis, there is an urgent need to develop novel peptide antibiotics that exhibit both antimicrobial and anti-inflammatory activities. Herein, we evaluated antimicrobial activity and anti-inflammatory activity of psacotheasin 2, one of the antimicrobial peptide candidates identified previously using an in silico analysis on the transcriptome of Psacothea hilaris. In addition to exhibiting antimicrobial activities against microorganisms without inducing hemolysis, psacotheasin 2 also decreased the nitric oxide production in lipopolysaccharide (LPS)-induced Raw264.7 cells. Moreover, ELISA and western blot analysis revealed that psacotheasin 2 reduced the expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Further, we found that psacotheasin 2 markedly reduced the expression levels of pro-inflammatory cytokines (IL-6 and IL-1β) by regulating mitogen-activated protein kinases (MAPKs) and nuclear factor-kB (NF-kB) signaling in LPS-induced Raw264.7 cells. We also confirmed that the binding of psacotheasin 2 to bacterial cell membranes occurs via a specific interaction with LPS. In mouse models of LPS-induced shock, psacotheasin 2 significantly enhanced the survival rate and recovered weight by attenuating pro-inflammatory cytokines. Thus, psacotheasin 2 could be a promising candidate as a peptide antiseptic agent.

A Novel Synthesis of Sex Pheromone from the Longicorn Beetle (Psacothea hilaris)

An asymmetric synthesis of (8Z,21R)-21-methylpentatriacont-8-ene, the sex pheromone of the yellow-spotted longicorn beetle Psacothea hilaris, has been achieved using Evan’s induction as the key step. Based on the asymmetric methylation product of chiral (R)-4-benzyl-1,3-oxazolidin-2-one, the carbon chain of the target molecule was assembled through a C5+C12+C11+C8 sequence. (2R)-4-(Benzyloxy)-2-methylbutan-1-ol, which can be obtained from γ-lactone following Evan’s protocol, was connected to a C12 alkyl group. The chiral methyl group remained the key moiety (97% ee). After another Wittig reaction and catalytic hydrogenation step, the designed key intermediate (13R)-13-methylheptacosan-1-ol was obtained. Finally, after oxidation and Wittig reaction, the synthesis of the target molecule was completed in 10 linear steps with an ultra-high overall yield of 36.2%.

Transcriptome Analysis of Psacothea hilaris: De Novo Assembly and Antimicrobial Peptide Prediction.

Simple SummaryMultidrug resistance for classical antibiotics is the major problem faced in clinical infections. Insects have antimicrobial peptides (AMPs) which are a part of the immune repertoire to protect it from microbial pathogens. AMPs are short chain protein molecules that function as antibacterial, antifungal, antiparasitic and antiviral factors. Insect AMPs are present in their hemolymph or produced in response to entry of pathogens and kill them to protect the host. The objective of this study is to identify novel AMPs from Psacothea hilaris, the yellow spotted long horned beetle. The beetle was immunized with bacteria and fungi and RNA was isolated. The RNA was processed and screened by in silico strategies to identify novel AMPs. We obtained one potential candidate which was tested to be effective against harmful bacteria and fungi. This will be useful in treating multidrug resistant microbes alone or in combination with antibiotics with future validations.Antimicrobial peptides (AMPs) are the frontline innate defense system evolutionarily preserved in insects to combat invading pathogens. These AMPs could serve as an alternative to classical antibiotics to overcome the burden of treating multidrug resistant bacteria. Psacotheasin, a knottin type AMP was isolated from Psacothea hilaris and shown to exhibit antimicrobial activity, especially against fungi through apoptosis mediated cell death. In this study, we aimed to identify novel probable AMPs from Psacothea hilaris, the yellow spotted longicorn beetle. The beetle was immunized with the two bacterial strains (E. coli and S. aureus), and the yeast strain C. albicans. After immunization, total RNA was isolated and sequenced in Illumina platform. Then, beetle transcriptome was de novo assembled and searched for putative AMPs with the known physiochemical features of the AMPs. A selection of AMP candidates were synthesized and tested for antimicrobial activity. Four peptides showed stronger activity against E. coli than the control AMP, melittin while one peptide showed similar activity against S. aureus. Moreover, four peptides and two peptides showed antifungal activity stronger than and similar to melittin, respectively. Collectively one peptide showed both antibacterial and antifungal activity superior to melittin; thus, it provides a potent antimicrobial peptide. All the peptides showed no hemolysis in all the tested concentrations. These results suggest that in silico mining of insects’ transcriptome could be a promising tool to obtain and optimize novel AMPs for human needs.

A method for rearing the yellow-spotted longicorn beetle, Psacothea hilaris (Coleoptera: Cerambycidae), to stabilize the last larval instar

The larvae of Psacothea hilaris (Pascoe) grown continuously under a long day in the laboratory pupate after the 4th, 5th, or 6th instar. This developmental polymorphism has complicated studies on the control of metamorphosis in P. hilaris. Since pupation in P. hilaris is known to be suppressed under a short day, a change in the photoperiod from a short to a long day with appropriate timing may assist in obtaining physiologically homogeneous larvae that pupate at the next molt. When the photoperiod was changed from 12-h light:12-h dark to 15-h light:9-h dark at the beginning of the 5th instar, the pupation rate at the next (5th) molt reached 90%, which was significantly higher than the proportion of larvae that pupated after the 5th instar when the larvae were grown continuously under a long day (52%). This rearing technique will expedite studies on the control of metamorphosis in P. hilaris.

First record of Rhoptrocentruspiceus Marshall (Hymenoptera, Braconidae, Doryctinae) as parasitoid of Psacotheahilarishilaris (Pascoe) (Coleoptera, Cerambycidae).

The species Rhoptrocentrus piceus Marshall (Hymenoptera: Braconidae) was reared from the larvae of the xylophagous beetle Psacothea hilaris hilaris (Pascoe) (Coleoptera: Cerambycidae), an exotic pest of Ficus and Morus species native to eastern Asia. It was recorded in the north of Italy in September 2005. This discovery is the first report of this species as parasitoids of the yellow spotted longicorn beetle all over the world.

Cloning, phylogeny, and expression analysis of the Broad-Complex gene in the longicorn beetle Psacothea hilaris.

Seven isoforms of Broad-Complex (PhBR-C), in which the sequence of the zinc finger domain differed (referred to as Z1, Z2, Z3, Z2/Z3, Z4, Z5/Z6, and Z6, respectively), were cloned from the yellow-spotted longicorn beetle Psacothea hilaris. The Z1–Z4 sequences were highly conserved among insect species. The Z5/Z6 isoform was aberrant in that it contained a premature stop codon. Z6 had previously only been detected in a hemimetabola, the German cockroach Blattella germanica. The presence of Z6 in P. hilaris, and not in other holometabolous model insects such as Drosophila melanogaster or Tribolium castaneum, suggests that Z6 was lost multiple times in holometabolous insects during the course of evolution. PhBR-C expression levels in the brain, salivary gland, and epidermis of larvae grown under different feeding regimens were subsequently investigated. PhBR-C expression levels increased in every tissue examined after the gut purge, and high expression levels were observed in prepupae. A low level of PhBR-C expression was continuously observed in the brain. An increase was noted in PhBR-C expression levels in the epidermis when 4th instar larvae were starved after 4 days of feeding, which induced precocious pupation. No significant changes were observed in expression levels in any tissues of larvae starved immediately after ecdysis into 4th instar, which did not grow and eventually died.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-3-539) contains supplementary material, which is available to authorized users.

Induction of ovipositional behavior on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE, by leaves of mulberry and several other Moraceous plants

Induction of ovipositional behavior on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE, by leaves of mulberry and several other Moraceous plants

Response of each days after emergence on Yellow-spotted longicorn beetle, Psacothea hilaris PASCOE to the attractant from mulberry stumps

Response of each days after emergence on Yellow-spotted longicorn beetle, Psacothea hilaris PASCOE to the attractant from mulberry stumps

Ecological studies on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Ecological studies on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Attractant effect of mulberry stumps and volatile substances emitted by them on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Attractant effect of mulberry stumps and volatile substances emitted by them on the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Microbial control of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE (Coleoptera; Cerambycidae), by an entomogenous fungus, Beauveria tenella

Microbial control of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE (Coleoptera; Cerambycidae), by an entomogenous fungus, Beauveria tenella

The plants for maturation feeding of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

The plants for maturation feeding of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

The discrimination of larval instars of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE by means of the head capsule width

The discrimination of larval instars of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE by means of the head capsule width

Improvement of artificial diet for mass-rearing of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Improvement of artificial diet for mass-rearing of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Effect of adult feeding on the sexual maturation and the oviposition of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

Effect of adult feeding on the sexual maturation and the oviposition of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE

The effect of DDVP against Cordyceps brongniartii, natural enemy of yellow-spotted longicorn beetle (Psacothea hilaris)

The effect of DDVP against Cordyceps brongniartii, natural enemy of yellow-spotted longicorn beetle (Psacothea hilaris)

Susceptibility of adults of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE (Coleoptera: Cerambycidae), reared with an artificial diet through their larval stage to an entmogenous fungus Beauveria brongniartii

Susceptibility of adults of the yellow-spotted longicorn beetle, Psacothea hilaris PASCOE (Coleoptera: Cerambycidae), reared with an artificial diet through their larval stage to an entmogenous fungus Beauveria brongniartii

Photoperiodic response of larvae of the yellow-spotted longicorn beetle Psacothea hilaris after removal of the stemmata

The role of the stemmata in photoperiodism has been examined in holometabolic insects, but the only reliable results in Coleoptera have been obtained in Leptocarabus kumagaii (Carabidae), the larvae of which do not respond to photoperiod without stemmata. In the present study, photoperiodism was examined in another coleopteran, Psacothea hilaris (Pascoe) (Cerambycidae), after surgical removal of the stemmata. Larvae reared under short-day conditions and transferred to long-day conditions on day 2 of the 5th instar pupated without further larval molts, whereas those continuously reared under short-day conditions underwent supernumerary molts and did not pupate. When the stemmata were removed on day 2 of the 5th instar, the larvae pupated under long-day conditions but did not do so under short-day conditions. However, under long-day conditions some underwent supernumerary molts before pupation. Larvae from which the sensilla trichodeum were removed showed a similar response to that of stemmata-deficient larvae, and larvae from which stemmata were removed at a younger stage (day 2 of the 4th instar) responded to photoperiod similarly to intact larvae. Thus, supernumerary molts under long-day conditions after removal of the stemmata were attributed to injury due to surgery, rather than a change in photoperiodic photoreception. Therefore, we conclude that larvae of P. hilaris show a photoperiodic response after removal of stemmata, in contrast to larvae of L. kumagaii.

Relationship between rapid cold-hardening and cold acclimation in the eggs of the yellow-spotted longicorn beetle, Psacothea hilaris

Rapid cold-hardening (RCH) and cold acclimation (ACC) were examined in eggs of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). When eggs incubated at 25 °C were transferred directly to conditions of −22 °C for 2 h, less than 30% survived, whereas exposure to 0 °C for 4 h prior to transfer to −22 °C increased survival to nearly 60%. The rapidly enhanced cold tolerance (RCH) was transient and lost rapidly after 1 h at 25 °C. Incubation at 15.5 °C for 9 days (ACC) also enhanced cold tolerance. Comparison of the cold tolerance of non-treated eggs and eggs pre-treated to give RCH, ACC, or ACC+RCH allowed the relationship between the two hardening processes to be determined. At a mild subzero temperature (−10 °C) an RCH effect was not detected, whereas only RCH is effective at the severest subzero temperature just above the SCP (−26 °C). At intermediate temperatures (−16, −22 and −25 °C), ACC and RCH enhanced survival in combination. Therefore, the two hardening processes have different physiological bases but operate concomitantly over a wide temperature range.

Molecular cloning and developmental expression of the gene encoding juvenile hormone esterase in the yellow-spotted longicorn beetle, Psacothea hilaris

Juvenile hormone (JH) plays a key role in the regulation of growth, development, diapause and reproduction in insects. The regulation of JH titers in the insect body is therefore crucial throughout postembryonic development. One of the major pathways of JH metabolism is degradation by a highly selective enzyme, juvenile hormone esterase (JHE). We obtained a full-length cDNA encoding JHE in Psacothea hilaris ( PhJHE). The complete PhJHE cDNA sequence is comprised of 1989 bp with an open reading frame of 1785 bp encoding 595 amino acid residues. The deduced protein sequence of PhJHE showed high homology with the Tenebrio molitor JHE (50% amino acid identity) and moderate homology with the Drosophila melanogaster JHE (34%). The PhJHE transcript was expressed mainly in the fat body. PhJHE transcript levels were low until day 3 of the 5th (final) larval instar, then steadily increased reaching a peak on day 13 (the prepupa stage), coinciding well with the peak hemolymph enzyme activity level. Sustained starvation of larvae after a period of feeding stimulated the expression of PhJHE mRNA while feeding the larvae with glucose downregulated its expression. These results are discussed with reference to the induction of precocious metamorphosis in this beetle by starvation.