大豆球蛋白对克氏原螯虾肠道菌群和抗菌相关基因的影响  

GLYCININ ON INTESTINAL MICROBIOTA AND ANTIMICROBIAL RELATED GENES IN RED SWAMP CRAYFISH(PROCAMBARUS CLARKI)

作　　者：王思如 矣林圆 杨慧君[1] 许乾 袁勇超[1] WANG Si-Ru;YI Lin-Yuan;YANG Hui-Jun;XU Qian;YUAN Yong-Chao(Key Lab of Freshwater Animal Breeding,Ministry of Agriculture,College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学水产学院,农业农村部淡水生物繁育重点实验室,武汉430070

出　　处：《水生生物学报》2024年第8期1300-1314,共15页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金面上项目(32273167);中央高校自主创新基金项目(2662022SCYJ003);湖北洪山实验室重大项目(2021hszd002)资助。

摘　　要：实验旨在探究不同水平的大豆球蛋白对克氏原螯虾(Procambarus clarkii)肠道菌群和抗菌相关基因的影响。在饲料中添加0 (G0组,作为对照组)、1.82%(GL组)、3.64%(GH1组)、5.46%(GH2组)和7.28%(GH3组)五种不同水平的大豆球蛋白饲喂克氏原螯虾(平均体重约为4.3 g) 4周。养殖实验结束后测定肠道微生物的组成和抗菌相关基因的表达。结果显示:(1)饲料中的大豆球蛋白对克氏原螯虾肠道菌群的Alpha多样性没有显著性影响,但改变了其肠道菌群组成。GL组Cloacibacterium、沙曼气单胞菌(Aeromonas sharmana)等菌群的相对丰度显著高于其他4组。与G0组相比,红杆菌属(Rhodobacter)、副球菌属(Paracoccus)、丙酸杆菌属(Propionicimonas)和生丝微菌属(Hyphomicrobium)等益生菌及亨氏柄杆菌(Caulobacter henricii)的相对丰度在GH2或GH3组中显著下降;而GH3组壤霉菌属(Agromyces)、巴氏微杆菌(Microbacterium barkeri)、假黄色单胞菌属(Pseudoxanthomonas)和军团杆菌属(Legionella)等潜在病原菌的相对丰度显著高于G0组。(2)随着大豆球蛋白水平的升高,各组肠道抗菌相关基因的表达水平呈现一次线性和二次响应的变化趋势。与G0组相比,GL组抗脂多糖因子(alf)、甲壳素(cru)和血蓝蛋白(hem)基因的表达水平显著下降;除组织蛋白酶-B (cst-b)外,GH1、GH2和GH3组其他抗菌相关基因的表达水平随着饲料中大豆球蛋白水平的升高而显著上升;GH2组cst-b的表达水平显著高于其余4组。(3)斯皮尔曼相关分析结果表明肠道菌群和抗菌相关基因之间存在显著的相关关系。cru、alf、C型凝集素(lec-c)、溶菌酶(lys)、白细胞介素增强因子结合子-2 (iebf-2)、hem、热休克蛋白-70 (hsp-70)和cst-b的表达水平与亨氏柄杆菌、副球菌属、红杆菌属、丙酸杆菌属、Desulfovibrio putealis、Thermomonas dokdonensis、沙曼气单胞菌和气单胞菌属、Chitinilyticum aquatile、台湾几丁质杆菌、Cloacibacterium和Glycinin found in soybean meal has been identified as an antinutrient factor,capable of impeding the growth and health of aquatic animals.However,limited research exists regarding the impact of glycinin on the intestinal microbiota and antimicrobial related genes in red swamp crayfish(Procambarus clarki).This study aimed to investigate the effects of five different glycinin levels on intestinal microbiota and antimicrobial related genes of crayfish.Crayfish with an average body weight of approximately 4.3 g subjected to diets containing five graded levels of glycinin(0,1.82%,3.64%,5.46%,and 7.28%)over a period of 4 weeks.The groups were designated based on the level of dietary glycinin as follows:GO(control group),GL,GH1,GH2,and GH3 group,respectively.Following the feeding trial,alterations and distinctions in the expression of intestinal microbiota and selected genes were evaluated.16S rRNA gene sequencing showed no significant difference in alpha diversity among the groups.However,the beta diversity was affected by different levels of dietary glycinin,indicating a shift in the composition of intestinal microbiota.Relative abundance analysis demonstrated a significant increase in Aeromonas sharmana and Cloacibacterium in the GL group compared to other groups.Conversely,Rhodobacter,Paracoccus,Propionicimonas,Hyphomicrobium,and Caulobacter henrici exhibited a significant decrease in abundance of GH2 or GH3 groups compared to the GO group.Additionally,Agromyce,Microbacterium barkeri,Pseudoxanthomonas,and Legionella were significantly enriched in the GH3 group.In addition,the expression of antimicrobial related genes displayed a complex trend of linear and quadratic responses with increasing levels of dietary glycinin.Initially,all selected genes were down-regulated,followed by upregulation,and finally down-regulation.Specifically,compared to GO group,the GL group showed a significant decrease in the expression of alf,cru,and hem.However,with increasing levels of dietary glycinin,the expression of iebf-2,hsp-70,alf,

关 键 词：大豆球蛋白 肠道菌群 抗菌相关基因 克氏原螯虾 

分 类 号：S966.1[农业科学—水产养殖]