| Atrijuglans hetaohei Yang?Lepidoptera:Gelechioidea?is considered as a major threat to walnut production.It is difficult to control the larvae in the field because the larval A.hetaohei can parasitize in the walnut fruits.For monitoring the occurrence period of adults for A.hetaohei,sex attractant is an important measure to improve the effectiveness of field control.Using sex pheromone attractant to monitor the occurrence period of adults is an important measure to improve the field control effect.Up to the present,the molecular mechanism of sex pheromone biosynthesis of A.hetaohei is not clear.By transcriptomic sequencing and analysis of the female ovipositor-pheromone glands?OV-PG?and male genitalia of A.hetaohei,we obtained a total of 92 putative genes related to sex pheromone biosynthesis.RT-PCR and Real-time PCR were used to detect the tissue expression of these candidate genes at the m RNA level.The results are as follows:1. Construction and analysis of unigene database of pheromone gland transcriptome of A.hetaoheiBy transcriptome sequencing of the pheromone glands of A.hetaohei with using Illumina hiseq?TM?4000 platform and assembling the reads using Trinity,68,697 unigenes were obtained,24,251 of the unigenes were matched to the entries in NCBI non-redundant?nr?protein database with a cut-off E-value of 10-5.The best matches were obtained for Amyelois transitella?22%?,followed by Bombyx mori?10%?.9,780 genes?35.75%?were annotated by GO classification.12416,12071,and 19315 genes were classified into three different subclasses,including cell component,molecular function,and biological process,respectively.The sequencing results provided an important basis for the study of metabolism pathway and gene function of the moth.2. Identification and expression of genes related to sex pheromone synthesis and transport of A.hetaoheiBased on the analysis of the terminal abdominal transcriptomic data of A.hetaohei,92genes related to the synthesis and transport of pheromones were screened and identified in 8different categories,including 13 fatty acid synthetases,15 desaturases,23 fatty acyl-coenzyme A reductases,18 acetyltransferases,9 alcohol oxidases,1 acetyl-Co A carboxylase,6 odorant-binding proteins,and 7 chemoreceptor proteins.Gene expression patterns were analyzed using q RT-PCR,which showed that there were differences in the expression of some candidate genes in different female and male tissues,in which 7 Ahet Des and 9 Ahet FAR showed strongly PG biased,whereas 5 Ahet Des and 10Ahet FAR were predominantly expressed in male genitalia.Phylogenetic analysis indicated that Ahet Des1,Ahet Des2,Ahet Des3,Ahet Des5,Ahet Des10,Ahet Des13,Ahet Des14,and Ahet FAR5 were clustered with genes known to have involved in pheromone synthesis in other lepidopteran species.In this study,the candidate genes related to sex pheromone synthesis and transport of A.hetaohei and their expression in different tissues of male and female adults were identified,which provided a theoretical basis for further study on the function of sex pheromone synthesis related genes,exploring the molecular mechanism of sex pheromone synthesis and clarifying the biosynthesis pathway of sex pheromone. |
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