| Atractomorpha sinensis I. Bolivar, a popular species of Pyrgomorphidae in Acridoidea, is widely distributed in China. Adults and nymphs mainly feed on stems and leaves, and also damage the growth and development of graminoid plants. At present, the research about A. sinensis is mainly focused on the aspects of morphology, population genetic diversity, gametogenesis and mitochondrial genome. However, the research and analysis about genome and transcriptome of A. sinensis have not been reported. Because of enormous large size of locust genome, it is expensive to sequence the genome and difficult to do bioinformatic analysis. Comparing with the genome, the scale of locusts’transcriptome is relatively small, and the study feasibility is appropriate. Besides, the research could study gene expression from the aspects of different types of cells and organisms in different developmental stages; it also reflects dynamic transcriptional level.In this study, transcriptome of A. sinensis (include nymph, female and male adults) had been sequenced deeply by Illumina HiSeqTM2000high-throughput sequencing platform, based on the assembling of the sequencing data, we did global analysis of A. sinensis by using bioinformatics method. By using the software edgeR, we found a lot of differentially expressed genes and analyzed their function, which provided a great quantity of information for studying development and gender regulation of A. sinensis. Finally, we constructed the mitochondrial genome transcriptional map of A. sinensis, and obtained large amounts of mitochondrial transcription information. The main conclusions of this study are as follows:1.The transcriptome sequencing of three samples (A. sinensis’s nymph, female and male adults) generated20Gb data. The nymph generated54,122,927pair reads, the female adult generated24,713,322pair reads, the male adult generated22,979,536pair reads. After assembling these data, we obtained60,382Unigene of A. sinensis, the average length of the all-Unigene is707bp, and GC content is43.02%. By using the Trinity software, we predicted the reading frame of the Unigene and obtained29,705CDS sequences, the average length of them is717bp, namely, the number of amino acid which are codoned by the PCGs are more than200. 2.We have annotated the60,382Unigene, through blasting against various database, including Nr, Nt, SwissProt, KEGQ COG and GO, totally27,325Unigenes can be matched with above databases, accounting for45%of the total Unigenes. Most of the Unigene sequences were matched with the GO database, a total of14,749, they were classed into three categories:cellular component, molecular function, biological process. Most of them were involved in molecular function, such as varieties of connection forms and catalytic activities. In cellular component, most genes were located in intracellular. In biological process, most genes were involved in physiological process.3.In KEGG analysis, the number of Unigenes which were involved in genetic information processing were4,391, including gene replication and repair, RNA transcription, protein translation and modification process, most genes participate in translation. The number of Unigenes which were involved in metabolism were4,003, including varieties of amino acid metabolism, carbohydrate metabolism, glycan biosynthesis and metabolism. In contrast, there were fewer genes associated with metabolic diseases. In addition,81Unigenes were annotated to the development process.4.We used the edgeR software to find the differentially expressed genes among the nymphs, female and male adults,9,069significantly differentially expressed genes (DEGs) were selected. Among them,5,925were up-regulated in nymphs, while3,144were up-regulated in adults. We did some functional annotation and analysis of differentially expressed genes,5,205Unigenes were assigned to GO classification,1,200involved in cellular component,1,518involved in biological process,2,487involved in molecular function, most genes of them focus on varieties of binding and activitity. There were2,382differentially expressed genes which involved in KEGG metabolic pathway, focusing on all kinds of metabolisms and genetic information process, especially genes which involved in insect hormone synthesis pathway expressed differentially, indicating that these genes play an important role in growth and development of A. sinensis.5. We found1,879differentially expressed genes between female adults and male adults,608were up-regulated in female,1,271were up-regulated in male.806DEGs were assigned to GO classification,115involved in cellular,448were annotated into molecular function,243involved in biological process.193differentially expressed genes were match with the KEGG metabolic pathway, among them, we found a gene, called CYP3A, it belongs to the CYP450enzyme system, and up-regulated in female adults, suggesting that it may be involved in the biosynthesis of the insect sex hormones.6. Through mitochondrial transcript mapping of A. sinensis, we found that the rRNA transcription efficiency is higher than the protein coding genes, indicating that rRNA plays an important role in ribosome assemble. Therefore, termination factor binds to the rRNA3’downstream to terminate the transcription activity, in order to maintain the high level of rRNA. The two pairs of overlapping genes, ATP8/ATP6, ND4/ND4L, are transcribed from a bicistronic mRNA respectively. A. sinensis has five large transcriptional units, they can be processed into mature transcripts through’tRNA punctuation model’. There are heterogeneity expressions among the protein-coding genes, it may be caused by the instability of transcript and transcriptional regulation mechanism. |
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