大银鱼早期发育阶段微生物群落特征及其与免疫和代谢的关联

doi:10.3785/j.issn.1008-9209.2023.03.281

浙江大学学报（农业与生命科学版）

2024, Vol. 50

Issue (1): 123-136 DOI: 10.3785/j.issn.1008-9209.2023.03.281

动物科学与动物医学

大银鱼早期发育阶段微生物群落特征及其与免疫和代谢的关联

周依帆¹(

),张希昭^1,²(

),周彦锋^1,²,徐东坡^1,²,王晨赫¹,郭世越¹,蒋书伦²,尤洋^1,²(

)

^1.南京农业大学无锡渔业学院，江苏无锡 214081
^2.中国水产科学研究院淡水渔业研究中心，农业农村部淡水渔业与种质资源利用重点实验室，江苏无锡 214081

Characteristics of microbial communities and their association with immunity and metabolism at the early developmental stage of Protosalanx chinensis

Yifan ZHOU¹(

),Xizhao ZHANG^1,²(

),Yanfeng ZHOU^1,²,Dongpo XU^1,²,Chenhe WANG¹,Shiyue GUO¹,Shulun JIANG²,Yang YOU^1,²(

)

^1.Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, Jiangsu, China
^2.Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, Jiangsu, China

全文: PDF(3592 KB) HTML

摘要：

为了解大银鱼早期发育过程中的微生物群落变化规律与特征，采集大银鱼胚胎期（心跳期，XT）、内源性营养期（采样时间为孵化后第1天，记为H₁）、混合性营养期（孵化后第4天，H₄）、开口摄食期（孵化后第7天，H₇）和外源性营养期（孵化后第10天，H₁₀）5个发育时期的样本。通过16S rRNA基因测序技术观察大银鱼早期发育过程中的微生物群落演替，尤其是摄食前后的微生物群落特征和关键菌属；并结合同批次样品的转录组数据，基于关联网络方法重点分析了与免疫和代谢相关的菌属。结果显示：大银鱼早期发育阶段各时期的β多样性存在显著差异（P＜0.001），其中，XT的优势菌群为黄杆菌属（Flavobacterium）和金黄杆菌属（Chryseobacterium）；胚后H₁时期的优势菌群为假单胞菌属（Pseudomonas）；H₄时期的主要菌群为黄杆菌属和假单胞菌属；H₇和H₁₀时期的主要菌群为弯曲杆菌属（Flectobacillus）和假单胞菌属。假单胞菌属在各时期中稳定存在。与多种免疫和代谢基因表达显著相关的是弯曲杆菌属等节点菌属。本研究首次获得了大银鱼早期发育阶段的微生物演替信息，同时，筛选到的优势菌属与节点菌属将为大银鱼苗种的规模化培育提供思路和参考。

关键词： 大银鱼; 早期发育; 微生物; 16S rRNA; 转录组

Abstract:

In order to understand the changes and characteristics of microbial communities during early development of Protosalanx chinensis, samples were collected at five developmental periods, which are the embryonic period (heartbeat stage, XT), the endogenous nutrition period (the first day after hatching, H₁), the mixed nutrition period (the fourth day after hatching, H₄), the open feeding period (the seventh day after hatching, H₇), and the exogenous nutrition period (the tenth day after hatching, H₁₀). Microbial community succession during early development was observed by 16S rRNA gene sequencing technology, especially the characteristics and key microbial genera before and after feeding. Combined with the transcriptome data of the same batch of samples, the microbial genera related to immunity and metabolism were analyzed based on the association network method. The results showed that there were significant differences in β diversity among the different periods at the early developmental stage (P＜0.001). The dominant bacteria in the XT period were Flavobacterium and Chryseobacterium, and the dominant bacterium in the H₁ period was Pseudomonas. The main bacteria in the H₄ period were Flavobacterium and Pseudomonas. The main bacteria in H₇ and H₁₀ periods were Flectobacillus and Pseudomonas. The abundance of Pseudomonas was stable at all developmental periods. Node bacteria such as Flectobacillus were significantly associated with the expression of various immune and metabolic genes. This study has obtained the microbial succession information of the early developmental stage of P. chinensis for the first time, and screened out the dominant bacteria and node bacteria, which will provide references and ideas for the scaled cultivation of P. chinensis fry.

Key words: Protosalanx chinensis early development microbiota 16S rRNA transcriptome

收稿日期: 2023-03-28 出版日期: 2024-03-01

CLC:

S917.4

基金资助: 国家现代农业产业技术体系项目(CARS-46);中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金资助项目(2023TD65);国家农业科学渔业资源环境滨湖观测实验站建设项目(NAES013FS05)

通讯作者: 张希昭,尤洋 E-mail: 18203889079@163.com;zhangxizhao@ffrc.cn;youy@ffrc.cn

作者简介: 周依帆（https://orcid.org/0009-0004-7090-7297），E-mail：18203889079@163.com

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引用本文:

周依帆,张希昭,周彦锋,徐东坡,王晨赫,郭世越,蒋书伦,尤洋. 大银鱼早期发育阶段微生物群落特征及其与免疫和代谢的关联[J]. 浙江大学学报（农业与生命科学版）, 2024, 50(1): 123-136.

Yifan ZHOU,Xizhao ZHANG,Yanfeng ZHOU,Dongpo XU,Chenhe WANG,Shiyue GUO,Shulun JIANG,Yang YOU. Characteristics of microbial communities and their association with immunity and metabolism at the early developmental stage of Protosalanx chinensis. Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 123-136.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2023.03.281 或 https://www.zjujournals.com/agr/CN/Y2024/V50/I1/123

表1 样本详情

图1 大银鱼早期发育阶段5个时期的示意图

图2 5个时期微生物群落的 α 多样性和 β 多样性分析A. α多样性分析的箱线图（使用Kruskal-Wallis方法检验各时期间差异的显著性）；B.基于Bray-Curtis距离的非度量多维尺度的β多样性分析。

图3 5个时期的微生物组成（门水平）1～3代表相同时期下的3个重复样本。图4同。

图4 5个时期的微生物组成（属水平）

图5 5个时期微生物群落的KEGG功能分析每个热图代表1个KEGG通路二级分类。每个色块代表每个时期的功能丰度平均值并以行为中心进行缩放，红色代表丰度高，蓝色代表丰度低。

图6 5个时期的微生物关联网络A.微生物模块化网络（每个节点代表1个ASV，节点大小代表ASV的平均相对丰度。红色线段表示节点间呈负相关，蓝色线段表示节点间呈正相关）；B.模块化网络的Hub节点分析（Hub得分越高，与其他节点的关联程度就越高）。

图7 5个时期微生物群落基因的KEGG注释及分类结果

图8 微生物群落与免疫通路相关基因的关联网络每个节点代表1个ASV或1个基因，节点的大小代表节点关联度。节点标签表示ASV所属的细菌属名。红色线段表示节点间呈负相关，蓝色线段表示节点间呈正相关。

图9 微生物群落与代谢通路相关基因的关联网络A.碳水化合物代谢、氨基酸代谢、脂质代谢相关基因的维恩图；B.微生物群落和代谢通路相关基因关联网络（每个节点代表1个ASV或基因，节点的大小代表节点关联度。节点标签表示细菌属名或基因名称。红色线段表示节点间呈负相关，蓝色线段表示节点间呈正相关）；C.节点关联度大于5的ASVs节点的热图；D.节点关联度大于5的基因节点的热图。

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