Identification and characterization of circadian clock genes in the head transcriptome of Conopomorpha sinensis Bradley.

Conopomorpha sinensis Bradley is the most detrimental pest to litchi and longan in China. Adult eclosion, locomotion, mating and oviposition of C. sinensis usually occur at night, regulated by a circadian rhythm. Nevertheless, our understanding of the linkages between adult circadian rhythms and clock genes remains inadequate. To address this gap, transcriptomic analysis was conducted on female and male heads (including antennae) of C. sinensis using the Illumina HiSeq 6000 platform to identify major circadian clock-related genes. The annotated sequences were analyzed by BLASTx, and candidate clock genes were classified based on conservation, predicted domain architectures, and phylogenetic analysis. The analysis revealed a higher conservation of these genes among the compared moths. Further, the expression profile analysis showed a significant spatiotemporal and circadian rhythmic accumulation of some clock genes during development. The candidate clock genes were predominantly expressed in the head, highlighting their crucial function in circadian rhythm regulation. Moreover, CsinPer, CsinTim1, and CsinCry1 displayed similar dynamic expressions with a peak expression level in the 4th age adults, suggesting their involvement in regulation of courtship and mating behaviors. The CsinPer and CsinTim1 mRNA oscillated strongly with a similar phase, containing a peak expression just before the female mating peak. This work will greatly contribute to understanding the circadian clock system of C. sinensis and provide valuable information for further studies of the molecular mechanisms involved in rhythmicity in fruit-boring pests.

Effect of diflubenzuron on the chitin biosynthesis pathway in Conopomorpha sinensis eggs.

Conopomorpha sinensis is the dominant borer pest of Litchi chinensis (litchi) and Euphoria longan (longan) in China. Control of C. sinensis is difficult because of its cryptic life habit; thus, an effective ovicide could be beneficial. The larvicidal effects of diflubenzuron (DFB) have been documented in many insect pest species. Therefore, DFB might be a useful ovicide to control C. sinensis. However, the detailed mode of action of DFB interference with insect molting and egg hatching is unclear. Thus, we studied alterations in expression of all genes potentially affected by DFB treatment using a transcriptome approach in 2-d-old C. sinensis eggs. Clean reads were assembled to generate 203 455 unigenes and 440 558 transcripts. A total of 4625 differently expressed genes, which included 2670 up-regulated and 1955 down-regulated unigenes, were identified. Chitin binding and chitin metabolic processes were among the most significant enriched pathways according to Gene Ontology analyses. Most of the genes that encode enzymes involved in the chitin biosynthesis pathway were unaffected, whereas genes that presumably encode cuticle proteins were up-regulated. Furthermore, altered expression patterns of 10 genes involved in the chitin biosynthesis pathway of C. sinensis embryos were observed in response to DFB treatment at different time points by quantitative reverse transcription polymerase chain reaction. We also observed abnormal development; there was reduced chitin content and modulated chitin distribution of newly hatched larvae, and altered egg hatching. Our findings illustrate an ovicidal effect of DFB on C. sinensis, and reveal more molecular consequences of DFB treatment on insects.

Sublethal Concentration of Beta-Cypermethrin Influences Fecundity and Mating Behavior of Carposina sasakii (Lepidoptera: Carposinidae) Adults.

The purpose of this study was to evaluate the sublethal effects of the beta-cypermethrin on calling behavior and fecundity of a major fruit-boring pest of apple, Carposina sasakii Matsumura. The mating rate, fertility (total number of eggs laid per female), and adult longevity of adults were remarkably decreased as compared with that in control when the adults were exposed to 10% lethal concentrations (LC10) of beta-cypermethrin (LC10♀ × LC10♂), and the age-specific survival rate (lx) was also negatively affected by sublethal beta-cypermethrin especially for the LC10â™€× LC10♂ mating combination. However, the age-specific fecundity (mx) was stimulated particularly in the combination of CKâ™€× LC10♂. Furthermore, the oviposition period was prolonged and the number of eggs was significantly increased for combinations of CKâ™€× LC10♂ and LC10â™€× CK♂. In the mating experiments, males in control or LC10-beta-cypermethrin treatments preferred to mate with females in control. It might be because of lower calling rate of female survivors treated with sublethal beta-cypermethrin. Our data indicate that treatment of beta-cypermethrin had a sublethal effect on the development and production of C. sasakii, and their mating behavior changes in surviving adults that may contribute to assortative mating.