Variation Of Microflora In The Digestive Tract And Prxs Gene Expression Analysis Of Apostichopus Japonicus Under High-temperature Stress

Apostichopus japonicus is nutritious. It is an important resource and it can enhance immunity, antifungal, antitumor, and anticoagulant functions of antifatigue. The sea cucumber enters the aestivation state due to high temperatures. Temperature in Fujian province is significantly higher than in the North of Fujian province, leading to southern culture of sea cucumber entered a period of aestivation in advance. Because of high temperature some sea cucumber will be death during aestivation, certain economic losses caused to production. This paper, the South under high temperature stress study on intestinal flora and culture of sea cucumber Prxs gene, explore the effect of high temperature on microbial flora and genes, for an in-depth study on basal metabolic and molecular mechanisms of sea cucumber in the summer provided a theoretical basis.We count statistics on intestinal flora of southern cultured by tablet-colony count method. The number of gastrointestinal flora 2.7×106 cfu/mL at room temperature(18℃). 26℃ high temperature bacteria two weeks after digestive tract bacteria decreased to 3.2×105 cfu/mL. The both of result is different signifitly(P<0.05). Isolation, cultivation and traditional method of combining modern molecular biological techniques, respectively, from the experimental group control group at room temperature and high temperature purification to 58 and 55 strains of bacteria. By constructing phylogenetic trees of the 16 S rRNA gene and Biolog automated identification system, digestive tract flora species group at room temperature of sea cucumber: alternate genus Pseudomonas(Pseudoalteromonas.sp), giant Oyster vibrios(Vibrio gigantis), Bacillus(Bacillus.sp), Siva of the genus(Shewanella.sp), marine Oceanobacter kriegii similar species of the Bacillus, Similar to Vibrio rotiferanus species of the genus Vibrio and Vibrio litoralis similar kind; high temperature group of sea cucumber apostichopus japonicus gastrointestinal flora species: Acinetobacter Johnson(Acinetobacter johnsonii), saprophytic staphylococci(Staphylococcus saprophyticus) and generic bacteria(Pantoea agglomerans).We design degenerate primers by iCODEHOP software, amplification of the Prxs gene fragment in the Middle, then cloning 3’ with RACE technology ends can be separately identified and KB 683 bp KB 475 bp sequences, with typical 2-Cys characteristic motif of the Prxs, "FYPLDFTFVCPTEI" and "GEVCPA".Bioinformatics software analysis revealed high homology with the Prx1 and Prx2 gene.With fluorescent quantitative PCR research the expression situation of purpose gene in different organization and in breathing tree organization under the high temperature stress. results displayed: Prx1 gene in breathing tree, and intestinal, and longitudinal muscle and the body wall organization are has expression, which breathing tree and intestinal in the of expression volume significantly above longitudinal muscle and body wall(P<0.05); Under high temperature stress the gene expression in breathing organization rose in the first 6-9 hour, and then down, after 48 h recover normal. Prx2 gene in the respiratory tree, intestines, also has expression in body and outer longitudinal muscle tissue, and the highest level of expression in the respiratory tree organization, substantially higher than the longitudinal muscle and body wall of intestines; Under high temperature stress, the gene expression in breathing organization rose in the first 3-6 hour(P<0.05), and then down, after 24 h recover normal.