Effects of Lactobacillus helveticus on Gene Expression in Intestinal Transcriptome of Loach Misgurnus anguillicaudatus
CUI Xiaolin1, DING Zhiwen1, MI Haoyu1, MENG Wenrong1, LIU Shu2, YANG Guang2, HOU Xiaoyue2, DONG Zhiguo1, FANG Yaowei2
1. College of Marine Science and Aquaculture, Jiangsu Ocean University, Lianyungang 222005, China; 2. College of Food Science and Engineering, Jiangsu Ocean University, Lianyungang 222005, China
Abstract:In order to explore the effect of Lactobacillus helveticus HML037 on the growth and disease resistance gene expression of loach Misgurnus anguillicaudatus, the intestinal tissues of the loach from control group (C) and L. helveticus HML037 addition group (L) were double ended sequenced by Illumina hiseq 2500 high-throughput sequencing platform. The original data were quality controlled and de novo assembled, and 40 877 unigenes with an average length of 1218 bp and 1942 bp of N50. DEGseq was used to analyze gene expression difference, with 604 differential genes, of which 326 genes were significantly up-regulated and 278 genes were significantly down-regulated. All the differentially expressed genes were enriched in five pathways by GO functional enrichment analysis. The oxidation-reduction process pathway was the most abundant, followed by the oxidoreductase activity pathway, and the differentially expressed genes involved in these pathways were all up-regulated. Enrichment analysis of KEGG pathway showed that the differentially expressed genes were mainly concentrated in peroxidase proliferation-activation receptor signaling pathways, fatty acid prolongation and fat digestion and absorption. The findings provide an important reference for the improvement of intestinal growth and disease resistance performance of the loach by using L. helveticus HML037 instead of antibiotics.
[1] FENG B, YI S V, LI R W, et al. Comparison of age and growth performance of diploid and tetraploid loach Misgurnus anguillicaudatus in the Yangtze River basin, China[J].Environmental Biology of Fishes,2017,100(7):815-828. [2] 陈景星,朱松泉.鳅科鱼类亚科的划分及其宗系发生的相互关系[J].动物分类学报,1984,9(2):201-208. [3] ZHOU X Y, YU Y Y, LI Y H, et al. Comparative analysis of mitochondrial genomes in distinct nuclear ploidy loach Misgurnus anguillicaudatus and its implications for polyploidy evolution[J].PLoS One,2014,9(3):e92033. [4] 艾炎军,邹叶茂,汤文浩,等.微生态制剂对泥鳅生长性能、体成分和免疫活性的影响[J].淡水渔业,2013,43(1):81-84. [5] 李源.谷氨酰胺对泥鳅生长和非特异性免疫的影响[D].雅安:四川农业大学,2013:3-4. [6] 孟霄鹏,孟阳,王悦,等.益生菌对凡纳滨对虾免疫功能及肠道菌群的影响[J].水产科学,2017,36(1):60-65. [7] 韩卓然,孙敬锋.水产动物益生菌的筛选及应用[J].水产科学,2016,35(1):93-98. [8] 李海兵,宋晓玲,李赟,等.水产动物益生菌研究进展[J].动物医学进展,2008,29(5):94-99. [9] KIM D, BECK B R, HEO S B, et al. Lactococcus lactis BFE920 activates the innate immune system of olive flounder (Paralichthys olivaceus), resulting in protection against Streptococcus iniae infection and enhancing feed efficiency and weight gain in large-scale field studies[J].Fish & Shellfish Immunology,2013,35(5):1585-1590. [10] LI K, ZHENG T L, TIAN Y, et al. Beneficial effects of Bacillus licheniformis on the intestinal microflora and immunity of the white shrimp, Litopenaeus vannamei[J].Biotechnology Letters,2007,29(4):525-530. [11] TSENG D Y, HO P L, HUANG S Y, et al. Enhancement of immunity and disease resistance in the white shrimp, Litopenaeus vannamei, by the probiotic, Bacillus subtilis E20[J].Fish & Shellfish Immunology,2009,26(2):339-344. [12] SHEN W Y, FU L L, LI W F, et al. Effect of dietary supplementation with Bacillus subtilis on the growth, performance, immune response and antioxidant activities of the shrimp (Litopenaeus vannamei)[J].Aquaculture Research,2010,41(11):1691-1698. [13] SUZER C, ÇOBAN D, KAMACI H O, et al. Lactobacillus spp.bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae:effects on growth performance and digestive enzyme activities[J].Aquaculture,2008,280(1/2/3/4):140-145. [14] VERSCHUERE L, ROMBAUT G, SORGELOOS P, et al. Probiotic bacteria as biological control agents in aquaculture[J].Microbiology and Molecular Biology Reviews,2000,64(4):655-671. [15] GILL H S. Probiotics to enhance anti-infective defences in the gastrointestinal tract[J].Best Practice & Research Clinical Gastroenterology,2003,17(5):755-773. [16] LLOYD A B, CUMMING R B, KENT R D. Prevention of Salmonella typhimurium infection in poultry by pretreatment of chickens and poults with intestinal extracts[J].Australian Veterinary Journal,1977,53(2):82-87. [17] GRABHERR M G, HAAS B J, YASSOUR M, et al. Full-length transcriptome assembly from RNA-Seq data without a reference genome[J].Nature Biotechnology,2011,29(7):644-652. [18] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR 2(-Dalta Delta C(T)) method[J].Methods,2001,25(4):402-408. [19] 吴凤霞,袁国华.趋化因子及其受体研究进展[J].川北医学院学报,2008,23(3):297-300. [20] MURPHY P M, BAGGIOLINI M, CHARO I F, et al. International union of pharmacology.ⅩⅫⅠ.Nomenclature for chemokine receptors[J].Pharmacological Reviews,2000,52(1):145-176. [21] BONO M R, ELGUETA R, SAUMA D, et al. The essential role of chemokines in the selective regulation of lymphocyte homing[J].Cytokine & Growth Factor Reviews,2007,18(1/2):33-43. [22] WANG W, LI M, FANG W H, et al. A predictive model for assessment of decontamination effects of lactic acid and chitosan used in combination on Vibrio parahaemolyticus in shrimps[J].International Journal of Food Microbiology,2013,167(2):124-130. [23] XIE C, MAO X Z, HUANG J J, et al. KOBAS 2.0:a web server for annotation and identification of enriched pathways and diseases[J].Nucleic Acids Research,2011,39(suppl_2):W316-W322. [24] 谢帝芝,王树启,游翠红,等.鱼类高度不饱和脂肪酸合成的影响因素及其机理[J].中国水产科学,2013,20(2):456-466. [25] 郁颖.四种不同生境鱼类脂肪酸去饱和酶和延长酶基因cDNA全序列的克隆与分子进化分析[D].广州:暨南大学,2011:1-9. [26] SPRECHER H. Metabolism of highly unsaturated n-3 and n-6 fatty acids[J].Biochimica et Biophysica Acta,2000,1486(2/3):219-231. [27] 戴超,王芳,房子恒,等.温度对蜕壳后期三疣梭子蟹能量代谢酶活力的影响[J].水产学报,2013,37(9):1334-1341. [28] 漆正堂,丁树哲,贺杰.丙酮酸脱氢酶活性调控在线粒体代谢与生长平衡中的作用[J].生命的化学,2008,28(3):307-310. [29] XUE L Y, QIAN K X, QIAN H Q, et al. Molecular cloning and characterization of the myostatin gene in croceine croaker, Pseudosciaena crocea[J].Molecular Biology Reports,2006,33(2):129-135. [30] XUE L Y, YANG Q Y, XIAO Z K, et al. Molecular characterization of myostatin in black seabream, Acanthopagrus schlegelii[J].DNA Sequence,2008,19(3):217-223. [31] BIGA P R, CAIN K D, HARDY R W, et al. Growth hormone differentially regulates muscle myostatin1 and-2 and increases circulating cortisol in rainbow trout (Oncorhynchus mykiss)[J].General and Comparative Endocrinology,2004,138(1):32-41.