克氏原螯虾内源性纤维素酶糖苷水解酶9基因家族鉴定及其表达分析  

Identification and Expression Analysis of Endogenous Cellulase Glycoside Hydrolase 9 Gene Family in Procambarus clarkii

作　　者：王霆辉 李旭光[2] 徐宇[2] 李佳佳[2] 王爱民[3] 明建华[1] 许志强[2] WANG Tinghui;LI Xuguang;XU Yu;LI Jiajia;WANG Aimin;MING Jianhua;XU Zhiqiang(College of Life Sciences,Huzhou University,Huzhou 313000,China;Key Laboratory of Genetic Breeding and Cultivation for Freshwater Crustacean,Ministry of Agriculture and Rural Affairs,Freshwater Fisheries Research Institute of Jiangsu Province,Nanjing 210017,China;College of Marine and Bioengineering,Yancheng Institute of Technology,Yancheng 224000,China)

机构地区：[1]湖州师范学院生命科学学院,湖州313000 [2]江苏省淡水水产研究所,农业农村部淡水虾蟹遗传育种与养殖重点实验室,南京210017 [3]盐城工学院海洋与生命科学院,盐城224000

出　　处：《动物营养学报》2023年第3期1985-1995,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：江苏省种业振兴“揭榜挂帅”项目(JBGS〔2021〕032);江苏现代农业重点项目(BE2020348);江苏省农业科技自主创新资金[CX(21)3176]。

摘　　要：本试验旨在分析鉴定克氏原螯虾内源性纤维素酶糖苷水解酶9(GH9)基因家族,研究其在投喂不同饲料时以及低氧-复氧胁迫下的表达特征,以期为克氏原螯虾对于植物性饲料的消化利用研究提供理论依据,同时为动物内源性纤维素酶高效利用研究积累数据资料。采用生物信息学与分子克隆相结合的策略,系统研究克氏原螯虾GH9基因家族的全基因组分布情况,获取克氏原螯虾GH9基因家族的结构特点、分布特征及其在投喂不同饲料(植物性饲料、动物性饲料、配合饲料)时以及低氧-复氧胁迫下的表达响应结果。结果显示:1)克氏原螯虾基因组中共有7个GH9基因家族成员(PcGH9-1、PcGH9-2、PcGH9-3、PcGH9-4、PcGH9-5、PcGH9-6、PcGH9-7),GH9基因家族成员的cDNA长度为1843~2354 bp,编码氨基酸残基数为510~585个,分布于LG23、LG39以及LG54等3条染色体上。克氏原螯虾GH9基因家族具有1个典型的Glyco hydro 9结构域,外显子数为7~14个,内含子数为6~13个。依据是否具有纤维素结合区(CBD),克氏原螯虾GH9基因家族可分为2个亚类,第Ⅰ亚类包括PcGH9-1、PcGH9-2和PcGH9-7,第Ⅱ亚类包括PcGH9-3、PcGH9-4、PcGH9-5、PcGH9-6。2)克氏原螯虾GH9基因家族主要在肝胰腺中高表达。3)克氏原螯虾肝胰腺中GH9基因家族表达量及β-1,4-内切葡聚糖酶活性在投喂植物性饲料时最高,在投喂动物性饲料时最低。4)低氧-复氧胁迫下,克氏原螯虾肝胰腺中PcGH9-1、PcGH9-2、PcGH9-3、PcGH9-4与PcGH9-5的表达量随着低氧胁迫时间延长而下降,PcGH9-6与PcGH9-7的表达量在低氧-复氧胁迫下变化较小;低氧-复氧胁迫下,β-1,4-内切葡聚糖酶活性随着低氧胁迫时间延长而显著下降(P<0.05)。综上可知,克氏原螯虾体内具有内源性纤维素酶GH9基因家族,GH9基因家族成员主要在肝胰腺中高表达。投喂植物性饲料可显著提升克氏原螯虾肝胰腺组织中GH9基因家族的表达及β-1,4-内切�This experiment was conducted to analyze and identify the glycoside hydrolase 9(GH9)gene family of endogenous cellulase in Procambarus clarkii,and to study its expression characteristics when fed different feeds or under hypoxia-reoxygenation stress,so as to provide theoretical basis for the study of plant feed digestion and utilization in P.clarkii,and to accumulate data for the efficient utilization of endogenous cellulase in animals.The whole genome distribution of the GH 9 gene family in P.clarkii was systematically studied by combining bioinformatics and molecular cloning strategies,and the structural characteristics,distribution characteristics of the GH9 gene family in P.clarkia and their expression response patterns when fed different feeds(plant feed,compound feed and animal feed)or under hypoxia-reoxygenation stress were obtained.The results showed as follows:1)there were seven GH 9 gene family members(PcGH9-1,PcGH9-2,PcGH9-3,PcGH9-4,PcGH9-5,PcGH9-6 and PcGH9-7)in the genome of P.clarkii.The cDNA length of GH9 gene family members was 1843 to 2354 bp,and the number of encoded amino acids was 490 to 583,which were distributed on the three chromosomes LG23,LG39 and LG54.The GH9 gene family of P.clarkii had one typical Glyco hydro 9 domains with exon number ranging from 7 to 14 and intron number ranging from 6 to 13.The GH 9 gene family of P.clarkii could be divided into two subclasses according to whether it has cellulose binding domain(CBD).The first subclass included PcGH9-1,PcGH9-2 and PcGH9-7,and the second subclass included PcGH9-3,PcGH9-4,PcGH9-5 and PcGH9-6.2)The GH9 gene family of P.clarkii was mainly highly expressed in hepatopancreas.3)The GH9 gene family expression level andβ-1,4-endoglucanase activity of P.clarkii were the highest when fed plant feed and the lowest when fed animal feed.4)Under hypoxic-reoxygenation stress,the expression levels of PcGH9-1,PcGH9-2,PcGH9-3,PcGH9-4 and PcGH9-5 in hepatopancreas of P.clarkii were decreased with the prolongation of hypoxic-reoxygenation stress,while

关 键 词：克氏原螯虾 GH9基因家族 Β-1 4-内切葡聚糖酶 不同饲料 低氧胁迫 

分 类 号：Q78[生物学—分子生物学]