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急性盐度胁迫对缢蛏肠道菌群结构及功能的影响

张紫娟 戴文芳 薛清刚 林志华

张紫娟,戴文芳,薛清刚,等. 急性盐度胁迫对缢蛏肠道菌群结构及功能的影响[J]. 海洋学报,2023,45(11):131–141 doi: 10.12284/hyxb2023146
引用本文: 张紫娟,戴文芳,薛清刚,等. 急性盐度胁迫对缢蛏肠道菌群结构及功能的影响[J]. 海洋学报,2023,45(11):131–141 doi: 10.12284/hyxb2023146
Zhang Zijuan,Dai Wenfang,Xue Qinggang, et al. Effects of acute salinity stress on the gut bacterial community structure and functional potentials of Sinonvacula constricta[J]. Haiyang Xuebao,2023, 45(11):131–141 doi: 10.12284/hyxb2023146
Citation: Zhang Zijuan,Dai Wenfang,Xue Qinggang, et al. Effects of acute salinity stress on the gut bacterial community structure and functional potentials of Sinonvacula constricta[J]. Haiyang Xuebao,2023, 45(11):131–141 doi: 10.12284/hyxb2023146

急性盐度胁迫对缢蛏肠道菌群结构及功能的影响

doi: 10.12284/hyxb2023146
基金项目: 浙江省自然科学基金项目(LQ22C190005);宁波市“泛3315计划”现代农业创新团队项目(2019−2023);浙江省“生物工程”一流学科学生创新计划项目(CX2022032)。
详细信息
    作者简介:

    张紫娟(1998—),女,四川省宜宾市人,主要从事海洋贝类微生物生态学研究。E-mail:zhangzijuan927@163.com

    通讯作者:

    戴文芳,助理研究员,主要从事水产养殖病害微生态防控技术研究。E-mail:daiwenfang@zwu.edu.cn

  • 中图分类号: Q938.8;P714+.5

Effects of acute salinity stress on the gut bacterial community structure and functional potentials of Sinonvacula constricta

  • 摘要: 盐度是水产动物肠道微生物稳态的重要影响因子,其对水产动物的生长和健康有着重要影响。缢蛏(Sinonovacula constricta)是一种埋栖类的滩涂养殖贝类,常受到盐度波动的影响。然而,盐度是否对缢蛏肠道组织及其肠道菌群造成不利影响尚不清楚。为此,本研究设置了低盐(5)、正常盐度(20)和高盐(35)3个不同的盐度胁迫组,分别在胁迫前(正常盐度20,0 d)和胁迫15 d后收集缢蛏样品,利用组织病理学、16S rRNA高通量测序技术以及PICRUSt2工具对各处理组样品的肠道组织结构、菌群构成及其潜在功能进行比较分析。结果显示,15 d的急性盐度胁迫对缢蛏肠道组织造成了不同程度的损害,表现为细胞空泡、组织坏死和绒毛杂乱等。在所有组中共鉴定到712个细菌操作分类单元(OTUs),其中共有OTUs占细菌总数的6.2%。低盐和高盐胁迫下缢蛏肠道主要优势菌分别隶属于γ-变形菌纲(γ-Proteobacteria)和α-变形菌纲(α-Proteobacteria)。急性盐度胁迫改变了肠道细菌α-多样性,且在低盐组中物种丰富度最低。低盐和高盐胁迫下缢蛏肠道细菌群落结构均发生了显著改变(p < 0.002),同时伴随肠道菌群种间互作的降低。3种盐度胁迫下缢蛏肠道菌群组装的生态过程主要是确定性过程,而在低盐胁迫下其确定性过程有所降低。功能预测结果表明,急性盐度胁迫显著改变了缢蛏肠道菌群介导的营养物质和能量代谢等相关的功能通路丰度,而与免疫相关的功能通路丰度在高盐组中显著升高。本研究结果表明,急性盐度胁迫可引起缢蛏肠道发生病理损伤并伴随肠道菌群及其潜在功能的明显改变,这些改变可能影响缢蛏的健康。
  • 图  1  3种盐度胁迫下缢蛏的肠组织切片

    A. 放大倍数×200;B. 放大倍数×400。a. 基底膜;b. 肌层;c. 肠腔;d. 绒毛;e. 杯状细胞。实心箭头:绒毛杂乱、缺损;空心箭头: 坏死;虚线箭头: 基底膜剥离;黑色方框: 空泡化

    Fig.  1  Gut tissue sections from Sinonvacula constricta under three salinity stress

    A. ×200 magnification; B. ×400 magnification. a. Basement membrane; b. muscle layers; c. gut lumen; d. gut villus; e. goblet cells. Solid filled arrow: cilia defects; hollow filled arrow: necrosis; dotted arrow: basement membrane stripped; square frame: vacuolation

    图  2  3种盐度胁迫下缢蛏肠道共有和特有的细菌OTU数韦恩图

    Fig.  2  Venn diagram of the number of shared and unique gut bacterial OTUs in Sinonvacula constricta underthree salinity stress

    图  3  3种盐度胁迫下缢蛏肠道优势细菌门(变形菌门归类到纲)(a)和细菌属(b)的平均相对丰度变化

    Fig.  3  Changes in the average relative abundances of dominant gut bacterial phyla (Proteobacteria is assigned to the class level) (a) and genus (b) in Sinonvacula constricta under three salinity stress

    图  4  3种盐度胁迫下缢蛏肠道细菌群落的α-多样性指数变化

    不同字母表示组间存在显著差异(p < 0.05)

    Fig.  4  Changes in the α-diversity indexes of gut bacterial communities in Sinonvacula constricta under three salinity stress

    Different letters indicate significant differences (p < 0.05) among groups

    图  5  非度量多维尺度分析(NMDS)展示3种盐度胁迫下缢蛏肠道细菌群落的结构差异

    Fig.  5  Non-metric multidimensional scaling analysis (NMDS) showing the difference in gut bacterial community structures among Sinonvacula constricta under three salinity stress

    图  6  3种盐度胁迫下缢蛏肠道细菌群落的拓扑特性参数变化

    Fig.  6  Changes in the topological characteristics of gut bacterial communities in Sinonvacula constricta under three salinity stress

    图  7  3种盐度胁迫下缢蛏肠道细菌群落的生态过程差异

    虚线表示两者同等重要。虚线以下的群落表示确定性过程控制群落组装,虚线以上的群落则表示随机性过程是主要的

    Fig.  7  Difference of ecological processes of gut bacterial communities in Sinonvacula constricta under three salinity stress

    The dashed line denotes equal roles for both. A community below the line indicates that determinism dominantly governs the community assembly, while a community above the line indicates that stochasticity is dominant

    图  8  热图展示3种盐度胁迫下缢蛏肠道细菌群落潜在功能途径的丰度(平方根转换)分布变化

    热图中颜色从蓝到红代表对应的功能途径丰度提高;不同盐度处理下的样本按照功能途径进行聚类

    Fig.  8  Heatmap showing the abundance (sqrt (x) transformed) distribution of potential functional pathways of gut bacterial communities in Sinonvacula constricta under three salinity stress

    The color from blue to red represents an increase in the abundance of the corresponding functional pathway; samples under different salinity stress were clustered according to functional pathways

    表  1  基于Bray-Curtis 距离比较3种盐度处理下缢蛏肠道细菌群落的整体差异性

    Tab.  1  Comparison of overall differences in gut bacterial communities among Sinonvacula constricta under three salinity stress based on Bray-Curtis distance

    组别 N0 N15 L15 H15
    R p R p R p R p
    N0
    N15 0.828 0.003
    L15 0.960 0.002 0.317 0.036
    H15 0.993 0.002 0.135 0.095 0.593 0.007
      注:加粗的数值表示差异显著(p < 0.05)。
    下载: 导出CSV

    表  2  基于非参数的多元置换方差分析不同盐度和胁迫时间对缢蛏肠道细菌群落的定量影响

    Tab.  2  Quantitative effects of different salinity and stress time on the variations in gut bacterial community among Sinonvacula constricta based on nonparametric permutational multivariate analysis of variance

    自由度 平方和 均方差 F模型 R2 p
    不同盐度 2 1.069 0.535 2.877 0.171 0.001
    胁迫时间 1 1.473 1.473 7.929 0.235 0.001
    残差 20 3.716 0.186 0.594
    总计 23 6.259 1
    下载: 导出CSV
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  • 收稿日期:  2023-04-06
  • 修回日期:  2023-07-10
  • 网络出版日期:  2023-10-31
  • 刊出日期:  2023-11-30

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