分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

基于转录组分析不同硒源调节黄颡鱼适应低温应激的信号通路及关键基因

  • 胡俊茹 ,
  • 袁夕宸 ,
  • 黄文 ,
  • 李国立 ,
  • 张羽帆 ,
  • 王国霞 ,
  • 赵红霞 ,
  • 黄燕华
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  • 1. 广东省农业科学院动物科学研究所, 农业农村部华南动物营养与饲料重点实验室, 广东省动物育种与营养公共实验室, 广东省畜禽育种与营养研究重点实验室, 广州 510640;
    2. 广州飞禧特生物科技有限公司, 广州 510640;
    3. 北京奥特奇生物制品有限公司, 北京 101499
胡俊茹(1979-),女,河北唐山人,副研究员,博士,从事水产动物营养与饲料研究。E-mail:hujunru1025@163.com

收稿日期: 2021-01-31

  网络出版日期: 2021-08-11

基金资助

广东省自然科学基金(2018A030313501);华南师范大学重点科技平台开放课题;佛山市财政专项资金-2021年度共建广东农业科技示范市项目

Analysis of Different Selenium Sources on Regulation of Signal Pathways and Key Genes of Pelteobrus fulvidraco under Low Temperature Stress Based on Transcriptome

  • HU Junru ,
  • YUAN Xichen ,
  • HUANG Wen ,
  • LI Guoli ,
  • ZHANG Yufan ,
  • WANG Guoxia ,
  • ZHAO Hongxia ,
  • HUANG Yanhua
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  • 1. Guangdong Key Laboratory of Animal Breeding and Nutrition, Key Laboratory of Animal Nutrition and Feed Science in South China of Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
    2. Guangzhou Fishtech Biotechnology Co., Ltd., Guangzhou 510640, China;
    3. Beijing Otqi Biological Products Co., Ltd., Beijing 101499, China

Received date: 2021-01-31

  Online published: 2021-08-11

Supported by

 

摘要

本试验基于转录组测序和实时荧光定量PCR (RT-qPCR)技术研究亚硒酸钠和酵母硒对低温(13℃)应激下黄颡鱼肝脏转录组和关键基因表达的影响,以期探讨不同硒源调节黄颡鱼适应低温应激的信号通路及关键基因。以5.7 g左右的黄颡鱼为研究对象,分别投喂添加亚硒酸钠(T1组)和酵母硒(T2组)的试验饲料,于养殖42 d后开展低温应激试验。结果显示:T2组黄颡鱼的增重率和存活率高于T1组,但差异不显著(P>0.05)。与亚硒酸钠相比,酵母硒显著提高了低温应激后黄颡鱼血清甘油三酯含量及抑制与产生超氧阴离子自由基能力(P<0.05),并显著降低了血清丙二醛含量(P<0.05)。肝脏转录组分析显示,T1组和T2组分别产生了140 574 638和139 455 508个高质量reads (clean reads),共筛选到147个差异表达基因,其中76个基因表达量上调,71个基因表达量下调,表达上调基因显著富集到蛋白质水解作用、水解酶活性、肽酶活性、肽链内切酶活性、羧肽酶活性、蛋白质代谢过程以及胰腺分泌、蛋白质消化与吸收、脂肪消化与吸收、花生四烯酸代谢、醚脂代谢等通路。这些通路中的硒蛋白M、磷脂酶A2、羧肽酶A1、类胰蛋白酶-1、胰凝乳蛋白酶样弹性蛋白酶家族成员2A等关键基因的表达量显著上调。通过RT-qPCR对上述差异表达基因进行验证,证实了转录组测序结果的可靠性。综上所述,摄食酵母硒为硒源饲料的黄颡鱼的生长性能、抗低温能力优于摄食亚硒酸钠为硒源饲料的黄颡鱼,酵母硒可能通过上调胰腺分泌、蛋白质消化与吸收、脂肪消化与吸收、花生四烯酸代谢、醚脂代谢等通路关键基因的表达提高黄颡鱼的抗低温应激的能力。

本文引用格式

胡俊茹 , 袁夕宸 , 黄文 , 李国立 , 张羽帆 , 王国霞 , 赵红霞 , 黄燕华 . 基于转录组分析不同硒源调节黄颡鱼适应低温应激的信号通路及关键基因[J]. 动物营养学报, 2021 , 33(8) : 4662 -4674 . DOI: 10.3969/j.issn.1006-267x.2021.08.045

Abstract

Transcriptome sequencing (RNA-seq) and real-time quantitative PCR (RT-qPCR) were used to study the effects of sodium selenite and selenium yeast on signaling pathways and key genes expression of Pelteobrus fulvidraco under low temperature stress. Pelteobrus fulvidraco (about 5.7 g) were fed diets supplemented with sodium selenite (T1 group) and yeast selenium (T2 group) for 42 d, respectively, and then to carry low temperature experiment. The results showed that the weight gain rate and survival rate in T2 group were higher than those in T1 group, but the differences were not significant (P>0.05). The serum triglyceride content and the ability to inhibit and produce superoxide anion free radical were significantly increased and the serum malondialdehyde (MDA) content was significantly decreased in T2 group compared with T1 group (P<0.05). Liver transcriptome analysis showed that 140 574 638 and 139 455 508 clean reads were produced in T1 and T2 groups, respectively. A total of 147 differentially expressed genes were selected, among which 76 genes were up-regulated and 71 genes were down-regulated. The up-regulated genes significantly enriched in proteolysis, hydrolase activity, endopeptidase activity, peptidase activity, carboxypeptidase activity, protein metabolic process, and pancreatic secretion, protein digestion and absorption, fat digestion and absorption, arachidonic acid metabolism and ether lipid metabolism pathways. The expression levels of key genes such as selenium protein M, phospholipase A2, carboxylpeptidase A1, trypsin-1-like, chymotrypsin 2A were significantly up-regulated in these pathways. These differentially expressed genes were verified by RT-qPCR, which confirmed the reliability of the transcriptome sequencing results. In conclusion, the growth performance and low temperature resistance of Pelteobrus fulvidraco used selenium yeast as selenium source are better than those used sodium selenite as selenium source. Selenium yeast can improve the resistance to low temperature stress of Pelteobrus fulvidraco by up-regulating the expression of key genes in pancreatic secretion, protein digestion and absorption, fat digestion and absorption, arachidonic acid metabolism, ether-lipid metabolism pathways.

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