| Cyrtotrachelus buqueti Guerin-Meneville is an important forestry drilling pest in South China.peck inside the sheaths of bamboo shoots to absorb nutrition using tubular beak,while its larvae eat bamboo shoots inside the tissue.C.buqueti has brought harm to various tuft and severely influenced the development of bamboo industry.So it is important to control the insect harmlessly by taking the most use of its preference as the forestry protection in the new development era.For those reasons,Headspace Solid Phase Micro-Extraction(HS-SPME)method,Gas Chromatograph-Mass Spectrometry(GC-MS),Electroantennogram Responses(EAG),Y-tube olfactometer and Illumina HiSeq were conducted to investigate the developmental transcriptional group differentially expresses,the semiochemicals of mate-seeking orientation behavior and the influences of combination of pheromone and host volatile,and then used the sex attractant to trap(catch)adults,which was an important way to realize the pests comprehensive pollution free control and resource utilization.The main results were follows:(1)Developmental transcriptomes of eggs,larvae from the first instar to the prepupalstage,pupae,and adults(females and males)from emergence to mating were built.Approximately 164.8 million clean reads were obtained and annotated into 108854 transcripts.These were assembled into 24338,21597,24798,21886,24642,and 83115 Unigenes for eggs,larvae,pupae,females,males,and the combined datasets.And 21058 Unigenes were exhibited in NR library,5890 Unigenes were exhibited in NT library,8063 Unigenes were exhibited in KO library,14748 Unigenes were exhibited in SwissProt library,17105 Unigenes were exhibited in PFAM library,17123 Unigenes were exhibited in GO library,10672 Unigenes were exhibited in KOG library.the transcriptome library of C.buqueti also showed that when those insects were in different developmental stage?(2)The volatile components of the C.buqueti unmated adults fecula were analyzed by Headspace Solid Phase Micro-extractio(HS-SPME)and Gas Chromatograph-Mass Spectrometry(GC-MS).The results showed that the volatile components of the unmated female fecula were classified to 7 species with 26 kinds,mainly terpene,hydrocarbon,alcohols,aldehydes,ketones,esters and Phenols,the unmated male fecula had 5 species with 18 kinds,mainly terpene,hydrocarbon,alcohols,esters and Phenols.It is preliminarily speculated that female fecula contain information compounds.(3)Combining GC-MS and EAG responses,the circadian rhythm of the volatile components of adults body surfaces of C.buqueti and EAG responses towards the volatile components in different time periods was analyzed.Analysis results indicate that there are 17 kinds of rhythmic volatile components on females of C.buqueti.while there are 15 rhythmic volatile components on male.It can be observed that in the time period of 10-12 AM,the rhythmic volatile components` extracting solution of female imagoes has the highest attracting rate for those non-mating male imagoes(P<0.05),which is consistent with the EAG responses,indicating that 10-12 AM is peak time of female to release pheromone,It is illustrating that the volatile components of male body surfaces may contain some semiochemicals,and the major secretion time period is 10-12 AM.(4)The electrophysiological and behavioral response of the C.buqueti adults to 15 kinds of cuticular volatiles were tested with EAG and a Y-tube olfactometer.The results showed that different cuticular volatiles had different effects on adults significantly,and adults performed different prefenrences.The strongest EAG response of unmated males is Dibutyl phthalate,while the strongest EAG response of unmated males is Benzothiazole.The unmated male adult's EAG response was generally higher than the unmated female adult.When Benzothiazole concentration is 100?g/?L?200?g/?L,the concentrationg d-Limonene,phenol and Dibutyl phthalate concentration is in 100?g/?L,the EAG response was significantly different between unmated males and unmated females.Besides,adults show different EAG responses towards the same compound with different concentration,the most intense EAG response of camphor and squalene is 200?g/?L and 1?g/?L respectively,and the most intense EAG response of other 13 compounds is 100?g/?L.In behavioral responses,Dibutyl phthalate,phenol and benzothiazole have relatively better attracting effect on unmated female and male,and Dibutyl phthalate have a significant difference in the attracting effect on unmated male(P<0.05).(5)The single volatile matter of(Dibutyl phthalate,phenol,2-Nonanone)with the concentration of 100?g/?L and standard substance compound mixture(Phenol : Ethylhexameter : 2-Nonanone : nonanal : Methyl pentadecanoate as 17:19:8:26:28)was added the three volatile organic compounds(linalool,indole,benzaldehyde)according to different ratios(9:1,7:3,5:5,3:7).The EAG response measure for different mixed components and the behavior measure of Y-tube olfactometer for four combinations with the most intense response towards EAG indicate that Benzaldehyde can improve the attractant ability to the male when added in the insect body mixture or the monomer compound,and it has a good synergistic effect.The best attracting effect for the male is the scaling of Dibutyl phthalate and benzaldehyde=5:5 or mixture and benzaldehyde =5:5(P<0.05).while the synergism of linalool and benzpyrole is not ideal,demonstrating that Benz aldehyde is expected to be used as the synergistic agent for sex pheromone of C.buqueti.In our study,developmental transcriptomes of different insect state of C.buqueti were constructed,and the volatile components of fecal and rhythmic volatile components of the male and female insects were extracted and analyzed.The daily peak period of the release of pheromones was found,and the volatile compounds such as dibutyl phthalate,phenol were identified as the information compounds,demonstrating that Benz aldehyde may have synergistic effect on male mate-seeking. |
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