Identification of Chemosensory Genes Based on the Antennal Transcriptomic Analysis of Plagiodera versicolora

Insects can sense surrounding chemical signals by their accurate chemosensory systems. This system plays a vital role in the life history of insects. Several gene families participate in chemosensory processes, including odorant receptors (ORs), ionotropic receptors (IRs), gustatory receptors (GRs), chemosensory proteins (CSPs), odorant binding proteins (OBPs), and sensory neuron membrane proteins (SNMPs). Plagiodera versicolora (Coleoptera: Chrysomelidae), is a leaf-eating forest pest found in salicaceous trees worldwide. In this study, a transcriptome analysis of male and female adult antennae in P. versicolora individuals was conducted, which identified a total of 98 candidate chemosensory genes including 40 ORs, 7 IRs, 13 GRs, 10 CSPs, 24 OBPs, and 4 SNMPs. Subsequently, the tissue expression profiles of 15 P. versicolora OBPs (PverOBPs) and 39 ORs (PverORs) were conducted by quantitative real-time PCR. The data showed that almost all PverOBPs and PverORs were highly expressed in the male and female antennae. In addition, several OBPs and ORs (PverOBP10, PverOBP12, PverOBP18, PverOR24, and PverOR35) had higher expression levels in female antennae than those in the male antennae, indicating that these genes may be taking part in some female-specific behaviors, such as find mates, oviposition site, etc. This study deeply promotes further understanding of the chemosensory system and functional studies of the chemoreception genes in P. versicolora.

Volatile metabolites of willows determining host discrimination by adult Plagiodera versicolora

Plagiodera versicolora Laicharting is a highly damaging leaf beetle foraging on willow leaves. In willow germplasm collections, observation has shown that Salix suchowensis Cheng was severely foraged by this leaf beetle while Salix triandra L. was damage free or only slightly damaged. Results of olfactometer bioassays show that the headspace volatiles from leaves of S. triandra significantly repelled adult beetles, suggesting that this species produces volatile repellents against P. versicolora. S. suchowensis had no effect on the beetles. Gas chromatography-mass spectrometry was carried out to profile the headspace volatile organic compounds and 23 compounds from leaves of the alternate species in significantly different concentrations were detected. The effects of 20 chemical analogs on host discrimination were examined. Olfactory response to these chemicals showed that o-cymene, a S. suchowensis specific constituent, significantly attracted adult P. versicolora. In contrast, cis-3-hexenyl acetate, a constituent concentrated more in S. triandra than in S. suchowensis, significantly repelled beetles. Mixing o-cymene and cis-3-hexenyl acetate in comparable concentrations as in the volatiles of S. suchowensis demonstrated that the latter could mask the attracting effect of the former, causing a neutral response by adult beetles to leaves of S. suchowensis against clean air. In addition, chemical analogs have the same effect as plants when resembling volatile organic compounds in real samples. Two volatile metabolites were detected triggering host discrimination by one of the most damaging insect pests to host and non-host willows. The two metabolites are of considerable potential for use as olfactory signs in managing the beetles.

Bacteria-Mediated RNA Interference for Management of Plagiodera versicolora (Coleoptera: Chrysomelidae)

RNA interference (RNAi) has emerged as a novel and feasible strategy for pest management. Methods for cost-effective production and stable delivery of double-stranded RNA (dsRNA) to the target insects are crucial for the wide application of RNAi for pest control. In this study, we tested the expression of dsRNA in RNaseIII-deficient Escherichia coli HT115 which was then fed to Plagiodera versicolora larvae, an insect pest of Salicaceae plants worldwide. By targeting six potential genes, including actin (ACT), signal recognition particle protein 54k (SRP54), heat shock protein 70 (HSC70), shibire (SHI), cactus (CACT), and soluble N-ethylmaleimide-sensitive fusion attachment proteins (SNAP), we found that feeding bacteria-expressed dsRNA successfully triggered the silencing of the five target genes tested and the suppression of ACT and SRP54 genes caused significant mortality. Our results suggest that the oral delivery of bacteria-expressed dsRNA is a potential alternative for the control of P. versicolora, and that ACT and SRP54 genes are the potent targets.

Contribution to the knowledge of development and harmfulness of imported willow leaf beetle (Plagiodera versicolora) (Coleoptera, Chrysomelidae)

The paper deals with the occurrence, development and harmfulness of imported willow leaf beetle (Plagiodera versicolora Laich.) in riparian and accompanying stands of the Svratka and Svitava rivers in the region of Brno. Salix fragilis L. and S. &times; rubens Schr. are the most damaged species there. Imagoes leave wintering places usually at the beginning of May. During about a 60-day period, they damage ca. 23.3 cm² leaves of S. fragilis and lay 250 to 730 (on average 539) eggs. The whole egg-laying consists of 16 to 48 (on average 35.5) groups with 6 to 26 (on average 15.2) eggs. The fecundity of females hatched in the laboratory is minimally twice lower. Larvae hatch after 5 to 8 (in the laboratory after 4.3) days and damage about 282 mm² leaves. The development from laying eggs until hatching imagoes of the <br />1^st generation takes about 21 (in the laboratory about 15) days. Imagoes of the 1^st generation occur on trees from the end of May to mid-August and die after completing their reproduction. Plagiodera versicolora creates 3 to 4 (in the laboratory 4 to 6) generations during a year. The small part of imagoes of the 2^nd generation, predominant part (or all) imagoes of the 3^rd generation and under conditions of a tetravoltine development all imagoes of the 4^th generation enter a diapause. In the course of the growing season, the food consumption of imagoes and larvae decreases and fecundity of females markedly decreases. Schizonotus sieboldi (Ratz.), Medina melania (Meig.) and M. luctuosa (Meig.) rank among main enemies of Plagiodera versicolora.