The complete mitochondrial genomes of two sibling species of camellia weevils (Coleoptera: Curculionidae) and patterns of Curculionini speciation.

Complete mitochondrial genomes contain large and diverse datasets for species delineation. To better understand the divergence of the two morphologically indistinguishable weevil species in Curculionini, we first sequenced and compared their complete mitochondrial genomes. The complete mitochondrial genomes of Curculio chinensis and Curculio sp. were 19,713 bp with an A + T content of 76.61% and 19,216 bp with an A + T content of 76.85%, respectively. All 37 of the typical mitochondrial genes were determined in both species. The 13 protein sequences of the two species shared high homology (about 90%) except for ATP8 (73.08%). The differences in secondary structure of ATP8 were the number of possible proteins and nucleic acid binding sites. There were 22 and 15 mismatched base-pairs in the tRNA secondary structures from C. chinensis and Curculio sp., respectively. Maximum Likelihood and Bayesian analyses indicated that Curculio sp. is a novel species closely related to C. chinensis. The divergence time estimation suggests that Cryptorhynchinae and Curculionini lines diverged in the Cenozoic Period, while C. chinensis and Curculio sp. diverged at 6.7079 (95% CI 5-13) Mya. This study demonstrates the utility of using complete mitochondrial gene sets for phylogenetic analysis and enhances our understanding of the genetic basis for the evolution of the Curculionini.

RNA-sequencing analysis of fungi-induced transcripts from the bamboo wireworm Melanotus cribricollis (Coleoptera: Elateridae) larvae.

Larvae of Melanotus cribricollis, feed on bamboo shoots and roots, causing serious damage to bamboo in Southern China. However, there is currently no effective control measure to limit the population of this underground pest. Previously, a new entomopathogenic fungal strain isolated from M. cribricollis larvae cadavers named Metarhizium pingshaense WP08 showed high pathogenic efficacy indoors, indicated that the fungus could be used as a bio-control measure. So far, the genetic backgrounds of both M. cribricollis and M. pingshaense WP08 were blank. Here, we analyzed the whole transcriptome of M. cribricollis larvae, infected with M. pingshaense WP08 or not, using high-throughput next generation sequencing technology. In addition, the transcriptome sequencing of M. pingshaense WP08 was also performed for data separation of those two non-model species. The reliability of the RNA-Seq data was also validated through qRT-PCR experiment. The de novo assembly, functional annotation, sequence comparison of four insect species, and analysis of DEGs, enriched pathways, GO terms and immune related candidate genes were operated. The results indicated that, multiple defense mechanisms of M. cribricollis larvae are initiated to protect against the more serious negative effects caused by fungal infection. To our knowledge, this was the first report of transcriptome analysis of Melanotus spp. infected with a fungus, and it could provide insights to further explore insect-fungi interaction mechanisms.

[Electrophysiological and behavioral responses of male Apamea apameoides (Draudt) (Lepidoptera: Noctuidae) to sex pheromone components].

The sex pheromone gland extracts collected from calling females of Apamea apameoides (Lepidoptera: Noctuidae) were analyzed with GC-MS, the electrophysiological and behavioral responses of the male adults to serial dilutions of sex pheromone components and their synthetic blends were investigated with Y-tube olfactometer in laboratory and in bamboo forest field. The results indicated that (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol were the functional components in the sex pheromone gland extracts. Electroantennogram (EAG) recordings showed that sex pheromone gland extracts, (Z)-11-hexadecenyl acetate, (Z)-11-hexadecen-1-ol and the mixture of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol all could elicit strong EAG responses, and the average EAG values increased with the increasing concentration of the sex pheromone. The blends of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol at the ratio of 57:43 elicited a higher EAG value than each singular component did. The results of behavioral assay by Y-tube olfactometer accorded with those of EAG responses on the whole, and the mixture of (Z)-11-hexadecenyl acetate and (Z)-11-hexadecen-1-ol at the ratio of 57:43 was more attractive than each component alone. In field tests with silicone rubber as pheromone dispensers (concentration = 10(4) ng · uL(-1)), the average number of male adults captured per trap by the mixture was (48.5 ± 6.7).

[Calling and mating behaviors of bamboo shoot borer Kumasia kumaso].

The calling and mating behaviors of Kumasia kumaso were studied in laboratory at (25 1) degrees C and (60 +/- 10) % RH under a cycle 14 L:10 D photo period, and the EAG response of male K. kumaso to female K. kumaso sex gland extracts was also tested. The calling and mating behaviors of K. kumaso could only be observed in scotophase. The females began calling at the first scotophase following emergence, and the peak of calling occurred during the second scotophase and decreased thereafter. The calling of the females commenced 0-4 h after dark, reached the maximum between the 5th and the 7th hour of the scotophase, and terminated during the last 1-2.5 h of the scotophase. Moth age had significant effects on the calling patterns. With increasing moth age, the onset time of calling advanced gradually, and the mean number of calling bouts as well as the calling length per bout had a gradual increase from calling day 1 to day 4 but decreased on day 5. The peak time and terminated time of calling advanced with increasing moth age. The mating of K. kumaso adults initiated during the first scotophase and terminated till the 5th scotophase following emergence, and the peak of mating was observed during the second scotophase. The peak of mating was observed 5.5-7.0 h after dark, and advanced with increasing moth age. Moth age affected the mean onset time of mating and the copulation duration significantly. With increasing moth age, the mean onset time of mating advanced, and the copulation duration decreased. Sex ratio had significant effects on the mating behavior as well. Treatment 2 female:1 male showed a significantly higher mating percentage, but an earlier onset time of mating and shorter copulation duration, as compared with treatment 1 female:1 male. The results of EAG test indicated that the male adults showed a significant EAG response to the sex gland extracts of 2-day-old virgin females.

Mud-puddling in the yellow-spined bamboo locust, Ceracris kiangsu (Oedipodidae: Orthoptera): does it detect and prefer salts or nitrogenous compounds from human urine?

C. kiangsu adults were observed visiting human urine, especially on hot summer days. The main chemicals in fresh human urine include inorganic salts and CO(NH(2))(2.) When human urine was incubated, NH(4)HCO(3) became the richest nitrogenous compound. The phagostimulants, repellents and attractants in urine were identified here. On the filter papers treated with fresh or incubated urine samples, the 5th instar nymphs and the adults started and continued gnawing around the edges, in contrast to the 3rd and the 4th instar nymphs. The consumed areas were dramatically greater on the filters treated with the urine samples incubated for 3-6 days. The feedings of both male and female adults were also stimulated by several urine-borne components such as NaCl, NaH(2)PO(4), Na(2)SO(4), KCl, NH(4)Cl and NH(4)HCO(3) but not by CO(NH(2))(2). Among them NaCl was the most powerful phagostimulant. The repelling, or attractive/arresting effects of CO(NH(2))(2) and NH(4)HCO(3) were also evaluated by a two-choice test. When exposed to water- and CO(NH(2))(2) solution-immersed filters simultaneously, the adults prefer to stay on water-immersed filter. In contrast, when provided water- and NH(4)HCO(3) solution-treated filters, the adults prefer to stay on NH(4)HCO(3) solution-treated filter. This demonstrated that CO(NH(2))(2) acted as a repellent and NH(4)HCO(3) as an attractant/arrestant. In the bamboo forest, similar feeding behavior was also elicited by NaCl, NH(4)HCO(3) but not by CO(NH(2))(2). Comparing to NaCl solution, a mixed solution of NaCl and CO(NH(2))(2) (1:1) significantly decreased the consumed area of the treated filters whereas a mixed solution of NaCl and NH(4)HCO(3) (1:1) dramatically increased the consumed area. These results demonstrated that the phagostimulatory effect by NaCl was reduced by CO(NH(2))(2) in fresh urine and was enhanced by NH(4)HCO(3) in incubated urine.

Trade-offs between constitutive and induced resistance in wild crucifers shown by a natural, but not an artificial, elicitor.

Constitutive and induced plant resistance against herbivores occurs throughout the plant kingdom, but little is known about the evolutionary relationship between these two types of resistances. We examined the relationships between constitutive and induced resistance to the diamondback moth, Plutella xylostella, in 11 wild species of crucifers, and analyzed the changes in volatiles associated with their expression in two species. We used larvae of P. xylostella and jasmonic acid (JA) as elicitors of the induced response. The level of resistance was estimated as the relative number of eggs laid on the plants by P. xylostella. Substantial variation in constitutive resistance was observed among the 11 crucifer species. When the plants were damaged by larvae, a negative correlation was found between constitutive and induced resistance. However, a positive correlation was detected between constitutive and induced resistance when the plants were treated by JA. The shift in resistance was associated with changes in the emission of volatiles. These results strongly suggest that (1) a trade-off occurs between constitutive and induced resistance in wild crucifers, and that (2) such a trade-off can be observed by treating the plants with a natural, but not an artificial, elicitor.