Identification And Screening Of Resistance-related Genes Of Macrobrachium Nipponense Exposed To Nitrite

Orient freshwater prawn(Macrobrachium nipponinse) natural distribution in China, Japan, Vietnam and Burma and other regions, widely distributed in fresh water all over the country. shrimp iIs Chinese most important economic freshwater. In the breeding water, nitrite content in freshwater shrimp is one of the growth and development of the most important indicators of quality, research and reveal the molecular mechanism of resistance shrimp nitrite stress, for the development of freshwater shrimp aquaculture industry, in particular to improve survival rate of breeding provide an important basis.In recent years, Illumina high-throughput sequencing technology has become a powerful tool for transcriptome analysis of each species identified, high-throughput sequencing studies in shrimp and hypoxia stress-related have been reported, but associated with a nitrite-based platform Illunina sequencing studies have not been reported. Nitrite concentration control in 10 mg / L(96 LC-50), and extracts nipponense hepatopancreas at 48 h when and under normal feeding conditions nipponense hepatopancreas high-throughput sequencing and alignment detection together. The results are as follows:(1) In this study, Japanese prawn breeding in the environment at low concentrations of nitrite. Nitrite concentration control in 10mg/L(96LC-50), and extracts nipponense hepatopancreas at 48 h when and under normal feeding conditions nipponense hepatopancreas high-throughput sequencing and alignment detection together. The results show that, after sequencing quality control, were obtained 34.68 Gb Clean Data, each sample Q30 base percentage not less than 86.33%, each sample Clean Data are up to 5.37 Gb, obtained after a total of 56,054 pieces Unigene De novo assembly. Where in the length of more than 1kb Unigene has 14,373 pieces. Unigene for functional annotation, including NR, Swiss-Prot, KEGG, COG, KOG, GO and Pfam database comparison, received a total of 19,022 pieces Unigene annotation results. Analysis of gene expression in each sample was. Differentially expressed genes clustering mode, functional annotation and enrichment analysis.(2) Selecting the 15 differentially expressed unigenes, by qRT-PCR validation of differential expression in the stress group and the normal group, showed fluorescence quantitative results with sequencing results express the same trend. After gene and bioinformatics analysis, select the relevant immune cell signaling pathways were studied.(3)Selecting the Guaninenucleotide-binding protein and Lysosome two key genes, two genes were detected in the liver and pancreas Japanese Prawns in different time periods(4h, 10 h, 24 h, 48 h, 72 h, 96h) expression, gene showed Lysosome the expression appears at 4h, were highest at 10 h. Gap junction gene expression levels peaked at 10 h and 48 h. And the expression of two genes in different freshwater shrimp hepatopancreas, gills and muscle of three different tissues 48 h, the results showed the highest expression of the two genes in the muscle, the result may be due to muscle nitrite high sensitivity causes.(4)Under stress conditions and freshwater shrimp hepatopancreas reared under normal conditions of shrimp hepatopancreas histological observation by electron microscopy with further increase in survival time of freshwater shrimp nitrite environment, hepatopancreas cells appeared necrosis, cell wall and basement membrane separation, severe wall rupture, indicating that low concentrations of nitrite Stress has destroyed the organizational structure of the liver pancreas, which in turn will affect the normal physiological function of liver and pancreas. Rough endoplasmic reticulum ribosome particles begin to fall off a lot, severe swelling of endoplasmic reticulum, mitochondria partial collapse of the mitochondrial membrane bilayer is dissolved, the emergence of electronic transparent area, and some tend to disintegration; microvilli deformation and a few broken, indicating that long effect nitrite hepatopancreas constitutes a greater damage and destruction, with the extension of treatment time, organelle damage heavier.In summary, this study nipponense transcriptome sequencing, transcriptome get a lot of information for transcriptome studies nipponense provide basic data. Was differentially expressed genes in the organizations of the differentially expressed genes were studied, suggesting that the key genes in nipponense resilience in the process, provide a theoretical basis for improving shrimp tolerance test nitrite.