Abstract
Cyrtotrachelus buqueti is an extremely harmful bamboo borer, and the larvae of this pest attack clumping bamboo shoots. Pheromone-binding proteins (PBPs) play an important role in identifying insect sex pheromones, but the C. buqueti genome is not readily available for PBP analysis. Developmental transcriptomes of eggs, larvae from the first instar to the prepupal stage, pupae, and adults (females and males) from emergence to mating were built by RNA sequencing (RNA-Seq) in the present study to establish a sequence background of C. buqueti to help understand PBPs. Approximately 164.8 million clean reads were obtained and annotated into 108,854 transcripts. These were assembled into 24,338, 21,597, 24,798, 21,886, 24,642, and 83,115 unigenes for eggs, larvae, pupae, females, males, and the combined datasets, respectively. Unigenes were annotated against NCBI non-redundant protein sequences, NCBI non-redundant nucleotide sequences, Gene Ontology (GO), Protein family, Clusters of Orthologous Groups of Proteins/ Clusters of Eukaryotic Orthologous Groups (KOG), Swiss-Prot, and KEGG Orthology databases. A total of 17,213 unigenes were annotated into 55 sub-categories belonging to three main GO categories; 10,672 unigenes were classified into 26 functional categories by KOG classification, and 8,063 unigenes were classified into five functional KEGG categories. RSEM software for RNA sequencing showed that 4,816, 3,176, 3,661, 2,898, 4,316, 8,019, 7,273, 5,922, 5,844, and 4,570 genes were differentially expressed between larvae and males, larvae and eggs, larvae and pupae, larvae and females, males and females, males and eggs, males and pupae, females and eggs, females and pupae, and eggs and pupae, respectively. Of these, three were confirmed to be significantly differentially expressed between larvae, females, and males. Furthermore, PBP Cbuq7577_g1 was highly expressed in the antenna of males. A comprehensive sequence resource of a desirable quality was constructed from developmental transcriptomes of C. buqueti eggs, larvae, pupae, and adults. This work enriches the genomic data of C. buqueti, and facilitates our understanding of its metamorphosis, development, and response to environmental change. The identified candidate PBP Cbuq7577_g1 might play a crucial role in identifying sex pheromones, and could be used as a targeted gene to control C. buqueti numbers by disrupting sex pheromone communication.
Highlights
The bamboo snout beetle Cyrtotrachelus buqueti Guerin-Meneville (Coleoptera: Curculionidae) is widely distributed in China, Vietnam, Burma, Thailand, and other Southeast Asian countries [1, 2]
This resulted in a total of 31,469,916, 36,773,825, 32,128,345, 33,070,448, and 31,434,121 clean reads in eggs, larvae, pupae, females, and males of C. buqueti, respectively
All clean reads were assembled into transcripts by Trinity software; the longest copy of redundant transcripts was regarded as a unigene [22, 24, 25]
Summary
The bamboo snout beetle Cyrtotrachelus buqueti Guerin-Meneville (Coleoptera: Curculionidae) is widely distributed in China, Vietnam, Burma, Thailand, and other Southeast Asian countries [1, 2]. In Sichuan Province, China, nearly 67,000 hm of forests are affected by C. buqueti every year. The damage rate is typically 50%–80%, in severe cases this may reach 100%. C. buqueti is a major forest pest, and the severity of the damage caused has become an important factor restricting the development of bamboo for paper making [4]. In April 2003, the State Forestry Administration of the People’s Republic of China released a list of 156 harmful forest organisms, which included C. buqueti among other pests and harmful mites
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