基于脾脏转录组筛选北京油鸡和广明白鸡抗热应激相关功能基因

doi:10.11843/j.issn.0366-6964.2023.05.013

摘要/Abstract

摘要： 旨在通过对北京油鸡(BY)和"广明2号"白羽肉鸡B系(广明白鸡,GM)在热应激和正常饲养条件下的脾脏转录组进行分析,鉴定两鸡种在应对热应激时的差异表达基因和信号通路,解析不同鸡种耐热性能差异的分子调控机制。试验动物采用相同饲养条件下的25日龄北京油鸡和广明白鸡,测定异嗜性粒细胞与淋巴细胞的比值(H/L)、超氧化物歧化酶(SOD)和总抗氧化能力(T-AOC)等表型数据,采集脾脏组织进行转录组测序(RNA-Seq)。基于RNA-Seq数据,结合表型进行加权基因共表达网络分析(WGCNA),筛选出与性状相关性较高的模块与基因。脾脏转录组测序结果表明,热应激组相比于对照组,北京油鸡中共鉴定到313个差异表达基因,其中169个上调表达,144个下调表达;广明白鸡中鉴定到235个差异表达基因,其中152个上调表达,83个下调表达。利用WGCNA分析,在北京油鸡中筛选出2个与H/L相关性较高的模块,在广明白鸡中筛选出4个与H/L和T-AOC强相关的模块。通过筛选模块中的Hub基因,发现两品种中均存在TRIM29基因,说明该基因可能在抗热应激方面发挥重要作用。本研究通过转录组分析,揭示了不同鸡种在热应激下基因的表达差异规律,并鉴定到与耐热相关的候选基因,为进一步研究鸡的耐热机制提供了新的思路和线索。

关键词: 北京油鸡, 广明白鸡, 热应激, 差异表达基因, 加权基因共表达网络分析

Abstract: The purpose of this study was to identify the differentially expressed genes (DEGs) and signaling pathways between Beijing You chicken and Guangming No.2 broiler line B(Guangming broiler) in response to heat stress, and to reveal the molecular regulatory mechanisms underlying heat tolerance of different chicken breeds by analyzing spleen transcriptomic data of two chicken breeds Beijing You chicken and Guangming No. 2 broiler line B (Guangming broiler) under heat stress and normal rearing conditions. The animals were 25-day-old Beijing You chickens and Guangming broilers under the same rearing managements. Heterophil to lymphocyte ratio (H/L), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) were measured, and spleen tissues were collected for RNA-Seq. WGCNA analysis was performed, in combination with gene expression matrix based on RNA-Seq data and the phenotypes, to screen the modules and genes with high correlation with the traits. Comparison of the transcriptome sequencing results between the control and heat stress groups showed that 313 DEGs in Beijing You chicken before and after heat stress, of which 169 genes were up-regulated and 144 genes were down-regulated; while 235 DEGs in Guangming broiler, of which 152 genes were up-regulated and 83 genes were down-regulated. Through WGCNA, 2 modules with high correlation with H/L were screened in Beijing You chicken, while 4 modules with strong correlation with H/L and T-AOC were screened in Guangming broiler. Screening of the Hub gene in the module revealed the presence of the TRIM29 gene in both breeds, which plays an important role under heat stress conditions. In this study, the differential patterns of gene expression in different chicken breeds under heat stress were revealed by RNA-Seq. The candidate genes related to heat tolerance were identified, which provided novel ideas and clues for further study of heat tolerance mechanism in chickens.

Key words: Beijing You chicken, Guangming broiler, heat stress, differentially expressed genes, WGCNA

中图分类号:

S831.2

王子渲, 王巧, 张锦, Astrid Lissette Barreto Sánchez, 郑麦青, 李庆贺, 崔焕先, 安炳星, 赵桂苹, 文杰, 李和刚. 基于脾脏转录组筛选北京油鸡和广明白鸡抗热应激相关功能基因[J]. 畜牧兽医学报, 2023, 54(5): 1905-1914.

WANG Zixuan, WANG Qiao, ZHANG Jin, Astrid Lissette Barreto Sánchez, ZHENG Maiqing, LI Qinghe, CUI Huanxian, AN Bingxing, ZHAO Guiping, WEN Jie, LI Hegang. Transcriptome Based Screening of Functional Genes Related to Heat Stress Resistance in Beijing You Chickens and Guangming Broilers[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1905-1914.

参考文献 32

[1]	VANDANA G D,SEJIAN V,LEES A M,et al.Heat stress and poultry production:impact and amelioration[J]. Int J Biometeorol,2021,65(2):163-179.
[2]	DEEB N,CAHANER A.Genotype-by-environment interaction with broiler genotypes differing in growth rate.3.Growth rate and water consumption of broiler progeny from weight-selected versus nonselected parents under normal and high ambient temperatures[J]. Poul Sci,2002,81(3):293-301.
[3]	KUMAR B,MANUJA A,AICH P.Stress and its impact on farm animals[J]. Front Biosci (Elite Ed), 2012, 4(5):1759-1767.
[4]	王继光,孙晓蕾,张辉,等.短期热应激对肉仔鸡生长性能及腺苷酸活化蛋白激酶信号通路的影响[J].动物营养学报,2019,31(12):5533-5540.WANG J G,SUN X L,ZHANG H,et al.Effects of short-time heat stress on growth performance and adenosine 5'-monophosphate activated protein kinase signal pathway of broiler chickens[J]. Chinese Journal of Animal Nutrition,2019,31(12):5533-5540.(in Chinese)
[5]	宋晓娜,任景乐,李爱军,等.夏季热应激对蛋鸡的影响及防控措施[J].中国家禽,2015,37(9):60-61.SONG X N, REN J L, LI A J, et al. Effects of summer heat stress on laying hens and preventive and control measures[J]. China Poultry, 2015, 37(9):60-61. (in Chinese)
[6]	TEDESCHI J N,KENNINGTON W J,BERRY O,et al.Increased expression of Hsp 70 and Hsp 90 mRNA as biomarkers of thermal stress in loggerhead turtle embryos (Caretta Caretta)[J]. J Therm Biol,2015, 47:42-50.
[7]	FELVER-GANT J N,MACK L A,DENNIS R L,et al.Genetic variations alter physiological responses following heat stress in 2 strains of laying hens[J]. Poult Sci,2012,91(7):1542-1551.
[8]	HAO Y,GU X H,WANG X L.Overexpression of heat shock protein 70 and its relationship to intestine under acute heat stress in broilers:1.Intestinal structure and digestive function[J]. Poult Sci,2012,91(4):781-789.
[9]	DRÖGE W.Free radicals in the physiological control of cell function[J]. Physiol Rev,2002,82(1):47-95.
[10]	OMAR R,PAPPOLLA M.Oxygen free radicals as inducers of heat shock protein synthesis in cultured human neuroblastoma cells:relevance to neurodegenerative disease[J]. Eur Arch Psychiatry Clin Neurosci, 1993, 242(5):262-267.
[11]	MASHALY M M,HENDRICKS III G L,KALAMA M A,et al.Effect of heat stress on production parameters and immune responses of commercial laying hens[J]. Poult Sci,2004,83(6):889-894.
[12]	ALTAN Ö,PABUÇCUOǦLU A,ALTAN A,et al.Effect of heat stress on oxidative stress,lipid peroxidation and some stress parameters in broilers[J]. Br Poult Sci,2003,44(4):545-550.
[13]	MCFARLANE J M,CURTIS S E.Multiple concurrent stressors in chicks.:3.Effects on plasma corticosterone and the heterophil:lymphocyte ratio[J]. Poult Sci,1989,68(4):522-527.
[14]	KANEHISA M,GOTO S.KEGG:kyoto encyclopedia of genes and genomes[J]. Nucleic Acids Res,2000, 28(1):27-30.
[15]	LANGFELDER P,HORVATH S.WGCNA:an R package for weighted correlation network analysis[J]. BMC Bioinformatics,2008,9:559.
[16]	HORVATH S,DONG J.Geometric interpretation of gene coexpression network analysis[J]. PLoS Comput Biol,2008,4(8):e1000117.
[17]	GHOLAMREZA Z,XI H,XI F,et al.How can heat stress affect chicken meat quality?-a review[J]. Poult Sci,2019,98(3):1551-1556.
[18]	LARA L J,ROSTAGNO M H.Impact of heat stress on poultry production[J]. Animals (Basel),2013, 3(2):356-369.
[19]	BAGATH M,KRISHNAN G,DEVARAJ C,et al.The impact of heat stress on the immune system in dairy cattle:a review[J]. Res Vet Sci,2019,126:94-102.
[20]	QUINTEIRO-FILHO W M,CALEFI A S,CRUZ D S G,et al.Heat stress decreases expression of the cytokines,avian β-defensins 4 and 6 and Toll-like receptor 2 in broiler chickens infected with Salmonella Enteritidis[J]. Vet Immunol Immunopathol,2017,186:19-28.
[21]	BARRETO SÁNCHEZ A L,WANG Q,THIAM M,et al.Liver transcriptome response to heat stress in Beijing you chickens and Guang Ming broilers[J]. Genes,2022,13(3):416.
[22]	GONZALEZ-GIL A M,ELIZONDO-MONTEMAYOR L.The role of exercise in the interplay between myokines,hepatokines,osteokines,adipokines,and modulation of inflammation for energy substrate redistribution and fat mass loss:a review[J]. Nutrients,2020,12(6):1899.
[23]	SINGH A K,CHAUBE B,ZHANG X B,et al.Hepatocyte-specific suppression of ANGPTL4 improves obesity-associated diabetes and mitigates atherosclerosis in mice[J]. J Clin Invest,2021,131(17):e140989.
[24]	WEE W K J,LOW Z S,OOI C K,et al.Single-cell analysis of skin immune cells reveals an Angptl4-ifi20b axis that regulates monocyte differentiation during wound healing[J]. Cell Death Dis,2022,13(2):180.
[25]	刘梅.急性热应激对肉仔鸡生长性能及脂肪代谢的影响[J].动物营养学报,2011,23(5):862-868.LIU M.Effects of acute heat stress on growth performance and lipid metabolism of broilers[J]. Chinese Journal of Animal Nutrition,2011,23(5):862-868.(in Chinese)
[26]	ALMADA A E,HORWITZ N,PRICE F D,et al.FOS licenses early events in stem cell activation driving skeletal muscle regeneration[J]. Cell Rep,2021,34(4):108656.
[27]	BAKIRI L,HAMACHER R,GRAÑA O,et al.Liver carcinogenesis by FOS-dependent inflammation and cholesterol dysregulation[J]. J Exp Med,2017,214(5):1387-1409.
[28]	JOHNSTONE C P,REINA R D,LILL A.Interpreting indices of physiological stress in free-living vertebrates[J]. J Comp Physiol B,2012,182(7):861-879.
[29]	DAVIS A K,MANEY D L,MAERZ J C.The use of leukocyte profiles to measure stress in vertebrates:a review for ecologists[J]. Funct Ecol,2008,22(5):760-772.
[30]	WANG J,ZHU B,WEN J,et al.Genome-wide association study and pathway analysis for heterophil/lymphocyte (H/L) ratio in chicken[J]. Genes (Basel),2020,11(9):1005.
[31]	XING J J,ZHANG A,ZHANG H,et al.TRIM29 promotes DNA virus infections by inhibiting innate immune response[J]. Nat Commun,2017,8(1):945.
[32]	XU M X,HU J X,ZHOU B X,et al.TRIM29 prevents hepatocellular carcinoma progression by inhibiting Wnt/β-catenin signaling pathway[J]. Acta Biochim Biophys Sin,2018,51(1):68-77.