岩扇贝原种和繁育后代肠道和内脏团细菌多样性分析
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摘要
为研究岩扇贝Crassadoma gigantean的生物学特性和消化代谢机能,采用传统培养方法分离培养加拿大原种岩扇贝内脏团(JN)和肠道(JC)及中国育种得到的子一代岩扇贝内脏团(YN)和肠道(YC)4种样品中的细菌,测定分离到的可培养细菌的16S rRNA基因并确定其分类地位;提取上述样品中的总DNA,运用Ion S5~(TM)XL高通量测序平台,对样品中细菌群落多样性进行分析。结果表明:用传统方法获得可培养菌株83株,选取代表菌31株测序并比对,其分别属于嗜冷杆菌属Psychrobacter、动性球菌属Planococcus、弧菌属Vibrio、假单胞菌属Pseudomonas、假交替单胞菌属Pseudoalteromonas、芽孢杆菌属Bacillus、库克菌属Kocuria和副球菌属Paracoccus;高通量测序结果显示,4种样品中有效序列群落结构可分为9个门,分别为变形菌门Proteobacteria、异常球菌-栖热菌门Deinococcus-Thermus、拟杆菌门Bacteroidetes、厚壁菌门Firmicutes、蓝细菌门Cyanobacteria、软壁菌门Tenericutes、放线菌门Actinobacteria、浮霉菌门Planctomycetes和疣微菌门Verrucomicrobia;加拿大原种岩扇贝肠道样品中的优势菌门为变形菌门(占80%),次优势菌门为拟杆菌门(占15%),内脏团样品中的优势菌门为变形菌门(占52%),次优势门为拟杆菌门(占18%);中国育种得到的子一代岩扇贝肠道样品中的优势菌门为变形菌门(占61%),次优势菌门为拟杆菌门(占26%),内脏团样品中的优势菌门为变形菌门(占73%),次优势菌门为拟杆菌门(占17%)。研究表明,岩扇贝的肠道和内脏团微生物具有丰富的多样性,加拿大原种岩扇贝和国内子一代岩扇贝的肠道和内脏团细菌群落结构存在一定差异。
In this paper, bacteria were isolated and cultured from viscera and digestive tract of Canadian rock scallop Crassadoma gigantea and the first generation of congener rock scallop from artificial breeding in China by the traditional method, and bacterial community diversity was analyzed by 16 S rRNA identification, and total DNA using Ion S5~(TM)XL high throughput sequencing platform. The results showed that 83 strains of culturable strains were found by the traditional method, and 31 strains of representative strains were sequenced and compared,including Psychrobacter, Planococcus, Vibrio,Pseudomonas, Pseudoalteromonas, Bacillus, Kocuria and Paracoccus. The high throughput sequencing indicated that the effective sequence community structure of the 4 samples was divided into 9 phyla: Proteobacteria, Deinococcus-Thermus, Bacteroidetes, Firmicutes, Cyanobacteria,Tenericutes, Actinobacteria, Planctomycetes, and Vercomicrobia, with dominant Proteobacteria in the intestines of Canadian rock scallops, accounting for 80%, and the subdominant Bacteroidetes, accounting for 15%. There were dominant Proteobacteria, accounting for 52%, and the subdominant Bacteroidetes, accounting for 18%, in the viscera of Canadian rock scallops. The dominant phylum Proteobacteria, accounting for 61%, and subdominant phylum Bacteroides, accounting for 26%, were observed in the intestine of the subgeneration rock scallops bred in domestic. The dominant phylum Proteobacteria, accounting for 73%, and the subdominant phylum Bacterioids, accounting for 17%, in the viscera in the first generation rock scallops. The findings indicated that there was rich in microorganism diversity in intestine and viscera of rock scallops, with to some extent, differences between Canadian rock scallop and subgeneration rock scallops bred in China.
In this paper, bacteria were isolated and cultured from viscera and digestive tract of Canadian rock scallop Crassadoma gigantea and the first generation of congener rock scallop from artificial breeding in China by the traditional method, and bacterial community diversity was analyzed by 16 S rRNA identification, and total DNA using Ion S5~(TM)XL high throughput sequencing platform. The results showed that 83 strains of culturable strains were found by the traditional method, and 31 strains of representative strains were sequenced and compared,including Psychrobacter, Planococcus, Vibrio,Pseudomonas, Pseudoalteromonas, Bacillus, Kocuria and Paracoccus. The high throughput sequencing indicated that the effective sequence community structure of the 4 samples was divided into 9 phyla: Proteobacteria, Deinococcus-Thermus, Bacteroidetes, Firmicutes, Cyanobacteria,Tenericutes, Actinobacteria, Planctomycetes, and Vercomicrobia, with dominant Proteobacteria in the intestines of Canadian rock scallops, accounting for 80%, and the subdominant Bacteroidetes, accounting for 15%. There were dominant Proteobacteria, accounting for 52%, and the subdominant Bacteroidetes, accounting for 18%, in the viscera of Canadian rock scallops. The dominant phylum Proteobacteria, accounting for 61%, and subdominant phylum Bacteroides, accounting for 26%, were observed in the intestine of the subgeneration rock scallops bred in domestic. The dominant phylum Proteobacteria, accounting for 73%, and the subdominant phylum Bacterioids, accounting for 17%, in the viscera in the first generation rock scallops. The findings indicated that there was rich in microorganism diversity in intestine and viscera of rock scallops, with to some extent, differences between Canadian rock scallop and subgeneration rock scallops bred in China.
引文
[1] 王紫竹.大连获准引进岩扇贝促进扇贝养殖业健康发展[N].中国海洋报,2017-08-10(02).
[2] Whyte J N C,Bourne N,Ginther N G.Biochemical and energy changes during embryogenesis in the rock scallop Crassadoma gigantea[J].Marine Biology,1990,106(2):239-244.
[3] Beitler M K.Toxicity of adductor muscles from the purple hinge rock scallop (Crassadoma gigantea) along the Pacific coast of North America[J].Toxicon,1991,29(7):889-894.
[4] Bendell L I.Survey of levels of cadmium in oysters,mussels,clams and scallops from the Pacific Northwest coast of Canada[J].Food Additives & Contaminants Part B:Surveillance,2009,2(2):131-139.
[5] Meyers T R,Burton T,Evans W,et al.Detection of viruses and virus-like particles in four species of wild and farmed bivalve molluscs in Alaska,USA,from 1987 to 2009[J].Diseases of Aquatic Organisms,2009,88(1):1-12.
[6] Culver C S,Richards J B,Page H M.Plasticity of attachment in the purple-hinge rock scallop,Crassadoma gigantea:implications for commercial culture[J].Aquaculture,2006,254(1-4):361-369.
[7] Tremblay I,Guderley H E,Himmelman J H.Swimming away or clamming up:the use of phasic and tonic adductor muscles during escape responses varies with shell morphology in scallops[J].Journal of Experimental Biology,2012,215(23):4131-4143.
[8] Canapa A,Barucca M,Marinelli A,et al.A molecular approach to the systematics of the Antarctic scallop Adamussium colbecki[J].Italian Journal of Zoology,1999,66(4):379-382.
[9] 王潇,曹善茂,印明昊,等.不同贝龄岩扇贝数量性征的相关性及通径分析[J].河北渔业,2018(2):23-28,38.
[10] 曹善茂,汪健,王谦,等.岩扇贝人工育苗的初步研究[J].大连海洋大学学报,2017,32(1):1-6.
[11] 曹善茂,梁伟锋,刘钢,等.不同生态因子对岩扇贝幼贝耗氧率和排氨率的影响[J].大连海洋大学学报,2017,32(3):280-286.
[12] 曹善茂,梁伟锋,汪健,等.岩扇贝幼贝滤食率的基础研究[J].大连海洋大学学报,2016,31(6):612-617.
[13] 曹善茂,王昊,陈炜,等.岩扇贝闭壳肌营养成分的分析及与中国3种扇贝的比较[J].大连海洋大学学报,2016,31(5):544-550.
[14] 王昊.岩扇贝与深海扇贝营养成分分析及与三种养殖扇贝的比较[D].大连:大连海洋大学,2016.
[15] 梅承,范硕,杨红.昆虫肠道微生物分离培养策略及研究进展[J].微生物学报,2018,58(6):985-994.
[16] Rappé M S,Giovannoni S J.The uncultured microbial majority[J].Annual Review of Microbiology,2003,57:369-394.
[17] 王保军,刘双江.环境微生物培养新技术的研究进展[J].微生物学通报,2013,40(1):6-17.
[18] 刘增新,柳学周,史宝,等.牙鲆(Paralichthys olivaceus)仔稚幼鱼肠道菌群结构比较分析[J].渔业科学进展,2017,38(1):111-119.
[19] 史秀清,张正,王印庚,等.大菱鲆(Scophthalmus maximus)仔稚鱼发育期消化道可培养细菌的菌群特征分析[J].渔业科学进展,2015,36(4):73-82.
[20] 张正,李彬,王印庚,等.基于高通量测序的池塘养殖半滑舌鳎消化道菌群的结构特征分析[J].水生生物学报,2015,39(1):38-45.
[21] 陈瑜,金雷,朱敬萍,等.双壳贝类微生物富集能力及环境水体对其影响分析[J].山东化工,2016,45(17):13-14,17.
[22] 王瑞旋,林韵锶,郭志勋,等.海南陵水企鹅珍珠贝肠道及其养殖水体中异养细菌耐药性研究[J].热带海洋学报,2013,32(6):96-100.
[23] 李佩佩,陈雪昌,张玉荣,等.海洋红细菌Lentibacter algarum胞外多糖的分离纯化及结构解析[J].生物工程学报,2014,30(3):455-463.
[24] 简书令,吴月红,买热帕提阿依,等.来源于中国南海的微生物新种乙醇速生杆菌(Celeribacter ethanolicus)多相分类研究(英文)[J].微生物学通报,2016,43(5):907-916.
[25] 孙雪莹.虾夷扇贝幼体及育苗池水体细菌群落动态及潜在益生菌筛选[D].大连:大连海洋大学,2016.
[26] 张皓.养殖环境微生物群落的动态变化及与水环境的互作[D].苏州:苏州大学,2015.
[27] 游龙,韩茵,张凯,等.芽孢杆菌产胞外酶的活性分析及其对凡纳滨对虾的作用[J].中国海洋大学学报:自然科学版,2018,48(3):80-87.
[2] Whyte J N C,Bourne N,Ginther N G.Biochemical and energy changes during embryogenesis in the rock scallop Crassadoma gigantea[J].Marine Biology,1990,106(2):239-244.
[3] Beitler M K.Toxicity of adductor muscles from the purple hinge rock scallop (Crassadoma gigantea) along the Pacific coast of North America[J].Toxicon,1991,29(7):889-894.
[4] Bendell L I.Survey of levels of cadmium in oysters,mussels,clams and scallops from the Pacific Northwest coast of Canada[J].Food Additives & Contaminants Part B:Surveillance,2009,2(2):131-139.
[5] Meyers T R,Burton T,Evans W,et al.Detection of viruses and virus-like particles in four species of wild and farmed bivalve molluscs in Alaska,USA,from 1987 to 2009[J].Diseases of Aquatic Organisms,2009,88(1):1-12.
[6] Culver C S,Richards J B,Page H M.Plasticity of attachment in the purple-hinge rock scallop,Crassadoma gigantea:implications for commercial culture[J].Aquaculture,2006,254(1-4):361-369.
[7] Tremblay I,Guderley H E,Himmelman J H.Swimming away or clamming up:the use of phasic and tonic adductor muscles during escape responses varies with shell morphology in scallops[J].Journal of Experimental Biology,2012,215(23):4131-4143.
[8] Canapa A,Barucca M,Marinelli A,et al.A molecular approach to the systematics of the Antarctic scallop Adamussium colbecki[J].Italian Journal of Zoology,1999,66(4):379-382.
[9] 王潇,曹善茂,印明昊,等.不同贝龄岩扇贝数量性征的相关性及通径分析[J].河北渔业,2018(2):23-28,38.
[10] 曹善茂,汪健,王谦,等.岩扇贝人工育苗的初步研究[J].大连海洋大学学报,2017,32(1):1-6.
[11] 曹善茂,梁伟锋,刘钢,等.不同生态因子对岩扇贝幼贝耗氧率和排氨率的影响[J].大连海洋大学学报,2017,32(3):280-286.
[12] 曹善茂,梁伟锋,汪健,等.岩扇贝幼贝滤食率的基础研究[J].大连海洋大学学报,2016,31(6):612-617.
[13] 曹善茂,王昊,陈炜,等.岩扇贝闭壳肌营养成分的分析及与中国3种扇贝的比较[J].大连海洋大学学报,2016,31(5):544-550.
[14] 王昊.岩扇贝与深海扇贝营养成分分析及与三种养殖扇贝的比较[D].大连:大连海洋大学,2016.
[15] 梅承,范硕,杨红.昆虫肠道微生物分离培养策略及研究进展[J].微生物学报,2018,58(6):985-994.
[16] Rappé M S,Giovannoni S J.The uncultured microbial majority[J].Annual Review of Microbiology,2003,57:369-394.
[17] 王保军,刘双江.环境微生物培养新技术的研究进展[J].微生物学通报,2013,40(1):6-17.
[18] 刘增新,柳学周,史宝,等.牙鲆(Paralichthys olivaceus)仔稚幼鱼肠道菌群结构比较分析[J].渔业科学进展,2017,38(1):111-119.
[19] 史秀清,张正,王印庚,等.大菱鲆(Scophthalmus maximus)仔稚鱼发育期消化道可培养细菌的菌群特征分析[J].渔业科学进展,2015,36(4):73-82.
[20] 张正,李彬,王印庚,等.基于高通量测序的池塘养殖半滑舌鳎消化道菌群的结构特征分析[J].水生生物学报,2015,39(1):38-45.
[21] 陈瑜,金雷,朱敬萍,等.双壳贝类微生物富集能力及环境水体对其影响分析[J].山东化工,2016,45(17):13-14,17.
[22] 王瑞旋,林韵锶,郭志勋,等.海南陵水企鹅珍珠贝肠道及其养殖水体中异养细菌耐药性研究[J].热带海洋学报,2013,32(6):96-100.
[23] 李佩佩,陈雪昌,张玉荣,等.海洋红细菌Lentibacter algarum胞外多糖的分离纯化及结构解析[J].生物工程学报,2014,30(3):455-463.
[24] 简书令,吴月红,买热帕提阿依,等.来源于中国南海的微生物新种乙醇速生杆菌(Celeribacter ethanolicus)多相分类研究(英文)[J].微生物学通报,2016,43(5):907-916.
[25] 孙雪莹.虾夷扇贝幼体及育苗池水体细菌群落动态及潜在益生菌筛选[D].大连:大连海洋大学,2016.
[26] 张皓.养殖环境微生物群落的动态变化及与水环境的互作[D].苏州:苏州大学,2015.
[27] 游龙,韩茵,张凯,等.芽孢杆菌产胞外酶的活性分析及其对凡纳滨对虾的作用[J].中国海洋大学学报:自然科学版,2018,48(3):80-87.