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Journal of Tropical Oceanography >

2023 , Vol. 42 >Issue 6: 127 - 136

DOI: https://doi.org/10.11978/2022264

Marine Biology

Study on species composition and distribution of fish eggs in Yongle Atoll and Dongdao Island by high-throughput sequencing technology

  • ZHAO Jinfa , 1 ,
  • LIU Yong 1, 2 ,
  • LI Chunhou 1 ,
  • WANG Teng , 1, 2 ,
  • SHI Juan 1 ,
  • XIAO Yayuan 1 ,
  • WU Peng 1 ,
  • SONG Xiaoyu 1
Expand
  • 1. South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, Observation and Research Station of Pearl River Estuary Ecosystem, Guangdong Province, Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Guangzhou 510300, China
  • 2. Scientific Observation and Research Station of Xisha Island Reef Fishery Ecosystem of Hainan Province, Key Laboratory of Efficient Utilization and Processing of Marine Fishery Resources of Hainan Province, Sanya Tropical Fisheries Research Institute, Sanya 572018, China
WANG Teng. email:

Copy editor: LIN Qiang

Received date: 2022-12-28

  Revised date: 2023-02-22

  Online published: 2023-02-27

Supported by

Hainan Natural Science Foundation(322CXTD530)

Hainan Natural Science Foundation(322MS153)

Special Financial Project of Ministry of Agriculture and Rural Affairs(NFZX2021)

Basic and Applied Basic Research Project of Guangdong Province(2019B030302004-05)

Central Public-interest Scientific Institution Basal Research Fund, CAFS(2023TD16)

Special Fund Project for Basic Scientific Research Business Expenses of Central Public Welfare Scientific Research Institutes of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences(2021SD04)

Special Fund Project for Basic Scientific Research Business Expenses of Central Public Welfare Scientific Research Institutes of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences(2019TS28)

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Abstract

Coral reef, known as the tropical rainforest in the ocean, was an important spawning ground, nursery and feeding ground for marine fish. Yongle atoll and Dongdao Island were typical coral reef ecosystems of the Xisha Islands. In order to better manage and protect the fishery and germplasm resources of the Xisha Islands, this study collected fish eggs from the Yongle atoll and the Dongdao Island in April 2021. High-throughput sequencing technology is used to classify and identify them, and the differences in species composition and community structure between the two regions were analyzed. The results showed that only fish eggs were collected in both areas, and a total of 27 species were identified, belonging to 4 orders, 16 families and 25 genera. The perciformes was dominant, accounting for 95.45% in the Yongle atoll and 85.71% in the Dongdao Island. The similarity index of roe species composition between the Yongle atoll and the Dongdao Island was significantly different, among which the similarity of order, family, genus and species was 25.00%, 37.50%, 32.00% and 33.33%, respectively. The occurrence frequency of fish eggs was 100%. The average density of fish eggs was 0.984 ind·m-3 in the Yongle atoll and 0.453 ind·m-3 in the Dongdao Island. The dominant species (Y≥0.02) were Decapterus sp., Katsuwonus pelamis and Halichoeres trimaculatus in Yongle atoll and Scarus forsteni, Chlorurus sordidus and Parupeneus multifasciatus in the Dongdao Island, with significant differences in the dominant species. In addition, there was an undocumented species (Ambassis sp.) found in this investigation. The species richness index (D) showed that the Yongle atoll (2.46) was higher than the Dongdao Island (1.87), while Shannon-Wiener diversity index (H′) and evenness index (J′) showed that the Yongle atoll (1.08, 0.35) was lower than the Dongdao Island (1.32, 0.50). The composition of fish eggs in both regions was dominated by reef-associated fish, and the relative abundance of pelagic-oceanic fish in the Yongle atoll (88.02%) was higher than that in the Dongdao Island (0.44%). The research results showed that both the Yongle atoll and the Dongdao Island are spawning grounds for marine fish, and the Yongle atoll has an open lagoon, which is more suitable as spawning grounds for pelagic-oceanic fish (especially tuna) than the Dongdao Island. Moreover, high-throughput sequencing technology can fill the gap of traditional morphological methods in the identification of fish eggs, and could be used as a supplementary means for the investigation of fishery resources. This study provides a scientific basis for the conservation and management of fishery and germplasm resources in the Yongle atoll and the Dongdao Island.

Key words： coral reef; lagoon; Xisha Islands; spawning ground; tunas

Cite this article

ZHAO Jinfa , LIU Yong , LI Chunhou , WANG Teng , SHI Juan , XIAO Yayuan , WU Peng , SONG Xiaoyu . Study on species composition and distribution of fish eggs in Yongle Atoll and Dongdao Island by high-throughput sequencing technology[J]. Journal of Tropical Oceanography, 2023 , 42(6) : 127 -136 . DOI: 10.11978/2022264

西沙群岛在我国南海诸岛中陆地面积最大, 海岛数量最多, 位于15°46′—17°08′N, 111°11′—112°54′E之间, 地处世界生物多样性最高的“珊瑚三角区”的北缘(李晓敏 等, 2020)。拥有我国最具代表性的珊瑚礁生态系统, 渔业资源丰富, 高于南海其他岛礁, 是我国最主要的热带渔场之一(李永振 等, 2007; 李永振 等, 2011)。西沙群岛属于热带季风气候, 年平均气温为26℃~28℃, 5—9月份水温最高, 月平均温度为29℃, 盐度较为稳定, 是众多海洋生物的产卵场, 对鱼类种群的繁衍生息具有十分重要的意义(Hou et al, 2020; 张婷 等, 2020)。鱼卵仔鱼被统称为鱼类浮游生物, 对海域鱼卵仔鱼的组成、分布、生物量等特征的研究可以更加全面的了解该海域的生态环境质量, 及时掌握鱼类群落和补充群体资源量状况, 对生态环境和渔业资源养护都具有重要的理论和实践意义(García, 2019; Zhang et al, 2022)。
由于缺乏明确的形态学特征, 以传统方法识别鱼卵是极其困难的, 目前可准确鉴定的鱼卵种类数仅占现存鱼种的百分之一(万瑞景 等, 2016; Chen et al, 2021)。为了准确鉴定鱼卵, 研究人员通常将卵培养到具有辨别性形态特征的幼体或成体阶段再进行鉴定, 但此等方法也较为耗时耗力(Gao et al, 2019)。随着分子技术的发展和成本的降低, 基于高通量测序技术已成为鉴定鱼卵的重要手段, 并已被广泛应用。Hofmann等(2017)使用分子技术与形态学方法相结合对三种重要经济鱼类进行鱼卵鉴定, 结果显示两种鉴定方法的一致性为96.2%, 证明了分子技术对鉴定鱼卵的有效性。Frantine-Silva等(2015)研究了巴拉那帕内马河的浮游鱼类, 并使用COI基因匹配鉴定出536个浮游鱼类样品中的99.25%, 证明了DNA条形码技术在识别鱼类浮游生物方面的功效, 以及使用鱼卵仔鱼作为生态监测工具的可能性。Choi等(2022)使用高通量测序技术对韩国水域上层的776个混合鱼卵进行了鉴定, 结果表明混合鱼卵的高通量测序可以有效的辅助物种的入侵检测。闫亚利(2019) 使用DNA条形码技术对西沙群岛琛航岛和七连屿的鱼卵仔鱼进行了研究, 结果表明DNA条形码技术可有效弥补传统形态学的不足之处。
本研究选取西沙群岛的永乐环礁和东岛的鱼卵仔鱼作为研究对象, 基于高通量测序技术对其进行研究, 旨在分析该海域鱼卵仔鱼的种类组成、资源分布和生物多样性等, 探讨高通量测序技术在鱼卵仔鱼鉴定方面的有效性, 为西沙群岛渔业资源的保护和管理提供基础信息。

1 数据和方法

1.1 研究区域

本研究于2021年4月在中国南海西沙群岛的永乐环礁和东岛进行鱼卵仔鱼调查采样(图1)。样品采集、保存和实验室分析测试等均按《海洋监测规范》(GB17378-2007)、《海洋调查规范》(GB/T 12763-2007)执行, 使用大型浮游生物网(网口内径80cm, 网目505μm)在调查海域进行水平拖拽采集。每站拖拽15min, 拖速2~3kn, 采集到的样品用浓度为95%乙醇溶液固定保存, 运回实验室进行分类鉴定与计数。
图1 西沙群岛调查采样分布图

Y1—Y9、D1—D4为采样站位点; 基于国家测绘地理信息局标准地图服务网站下载的审图号为GS(2019)1825号的标准地图制作

Fig. 1 Sampling distribution in the Xisha Islands (Y1-Y9, D1-D4 are the sampling points)

1.2 鱼卵仔鱼计数与鉴定

1.2.1 形态学方法

采集到的鱼类早期资源在解剖镜下进行初步处理, 将鱼卵分别进行分类与计数, 并根据卵径大小和胚体形态等典型形态特征进行初步分类(张俊彬 等, 2003; 肖瑜璋 等, 2013; 胡兴坤 等, 2022), 再通过COI分子标记进行精确种类鉴定。由于采集时在海上不便观察, 因此对样品全部采用95%乙醇溶液保存(郭琴 等, 2020), 从而致使鱼卵蛋白质变性, 无法直接观察到鱼卵的发育期, 因此, 本研究未鉴定鱼类早期资源的发育期。

1.2.2 分子方法

对95%乙醇溶液保存的鱼卵样品进行分子实验, 每个站位一个混合样, 并均设置三组平行, 采用通用型动物基因组DNA提取试剂盒(擎科生物科技有限公司)提取基因组DNA。之后选用鱼类通用引物(COI-L: TCAACCAACCACAAAGACATTGGCAC; COI-H: TAGACTTCTGGGTGGCCAAAGAATCA)对样品进行扩增(Ward et al, 2005)。PCR采用2×Es Taq MasterMix (Dye), 反应体系 (总体积为30µL): 2×Es Taq MasterMix (Dye) + dNTPs (15µL)、模板DNA (4.8µL)和上下游引物各1.2µL(10µL), 最后用ddH2O将体系补齐。PCR反应程序: 预变性94℃ 2min; 变性94℃ 30s, 10个循环; 退火55℃ 30s, 10个循环; 延伸72℃ 30s, 10个循环; 终延伸72℃ 2min, 于10℃保存。使用ddH2O为模板进行PCR阴性对照, 以评估PCR扩增中是否出现污染(Stoeckle et al, 2022)。最后经2%琼脂糖凝胶电泳检测PCR产物, 选取包含目的片段的产物, 于上海凌恩生物科技有限公司使用Illumina NovaSeq 6000测序平台进行测序(马红英 等, 2021)。
Illumina PE250测序序列首先根据barcode得到所有样品的有效序列; 然后对reads的质量进行质控过滤; 接着根据PE reads之间的overlap关系, 将成对的reads拼接成一条序列; 最后按照barcode和引物序列拆分得到每个样本的优质序列, 并在过程中根据正反barcode和引物方向校正序列方向以及去除嵌合体。按序列相似性≥97%进行OTU聚类分析(舒璐 等, 2020)后将OTU代表序列与Mito Fish数据库进行比对、分类注释, 并以错配低于2个碱基匹配到唯一的物种序列, 各采样点重复样品中匹配到物种序列数(read counts)取平均值处理, 并排除未匹配到的物种序列(Sales et al, 2020)。

1.3 数据处理

鱼卵物种丰度(A)的计算公式如下:
$A=\frac{N}{V}=\frac{N}{SL}=\frac{N}{Svt}$
式中 A为物种丰度(ind.·m-3), N为鱼卵个体数量(ind.), V为滤水量(m3), S为网口面积(m2), L为拖网作业距离(m), v为拖网速度(m·h-1), t为拖网的时间(h)(Song et al, 2019)。
鱼卵优势度(Y)的计算公式如下:
$Y=\frac{{{a}_{i}}}{A}{{f}_{i}}$
式中ai为第i个种类的物种密度(ind.·m-3), fi为第i个物种的出现频率, 优势种以Y≥0.02为划分标准(Xu et al, 2016)。
Jaccard相似性系数(Js)的计算公式如下:
${{J}_{\text{s}}}=\frac{c}{a+b-c}$
式中a、b分别为地点a、b的鱼卵种类数或目、科、属的数量; c为两地调查的鱼卵共有种类数或者目、科、属的数量。
物种丰富度指数(D)的计算公式如下:
$D=\frac{S-1}{\text{ln}N}$
香农-威纳多样性指数(H′)的计算公式如下:
$H'=-\sum\nolimits_{i=1}^{S}{{{P}_{i}}\text{ln}{{P}_{i}}}$
均匀度指数(J′)的计算公式如下:
$J'=\frac{H'}{\ln S}$
式(4)—(6)中, S为种类数, N为总个体数, Pi为第i种个体数占总个体数的比例。
按不同生态类型(礁栖鱼类、大洋性中上层鱼类和深海鱼类)将鉴定的鱼卵种类进行分类(陈真然, 1979)。
使用Primer 5.0对不同站点的鱼卵种类分布进行聚类分析, 来比较不同位置鱼卵群落结构的相对丰度和变化程度。采用Excel 2019、SPSS 24.0、Origin 2021进行数据统计分析, 采用ArcMap 10.6、Origin 2021、Photoshop 2019进行图片处理。

2 结果

2.1 鱼卵资源量特征

本次调查样品采集到鱼卵6204粒。其中, 永乐环礁5151粒, 平均密度为0.984ind.·m-3, 东岛1053粒, 平均密度为0.453ind.·m-3。永乐环礁鱼卵资源量普遍高于东岛, 永乐环礁鱼卵总平均密度为东岛的2.17倍。根据永乐环礁和东岛的鱼卵数量站位分布(图2a、c)可知, 在四月份的鱼卵调查中, 鱼卵的出现频率为100%, 每个站点平均为477粒。
图2 永乐环礁和东岛鱼卵物种和生物量分布

a. 永乐环礁鱼卵生物量; b. 永乐环礁鱼卵种类; c. 东岛鱼卵生物量; d. 东岛鱼卵种类

Fig. 2 Distribution of fish egg species and biomass in the Yongle atoll and Dongdao Island. (a) Biomass of eggs in the Yongle atoll; (b) biomass of species in the Yongle atoll; (c) number of eggs in the Dongdao Island; (d) number of species in the Dongdao Island

2.2 鱼卵种类组成

两区域共鉴定到27种鱼卵, 隶属于4目16科25属。以鲈形目(Perciformes)最多, 占总物种数的88.89%, 其余为刺鱼目(Gasterosteiformes)、鼠鱚目(Gonorhynchiformes)和鲀形目(Tetraodontiformes)均为1科1种。其中有5种仅鉴定到属, 分别为带鱼属未定种(Trichiurus sp.)、金枪鱼属未定种(Thunnus sp.)、圆鲹属未定种(Decapterus sp.)、双边鱼属未定种(Ambassis sp.)和前角鲀属未定种(Pervagor sp.)。永乐环礁鱼卵种类数明显高于东岛(图2b、d)。永乐环礁共22种, 隶属于2目11科20属, 其中, 鲈形目(Perciformes)占绝对优势, 占总数的95.45%, 其次为鼠鱚目(Gonorhynchiformes)仅有1种。东岛共14种, 隶属于3目11科13属, 鲈形目(Perciformes)最多, 占总数的85.71%, 刺鱼目(Gasterosteiformes)和鲀形目(Tetraodontiformes)均为1科1种(表1)。
表1 永乐环礁与东岛鱼卵种类组成

Tab. 1 Species composition of fish eggs and larvae in the Yongle atoll and Dongdao Island

目 科 属 种 生态类型 永乐环礁 东岛
相对丰度/% Y 相对丰度/% Y
刺鱼目
Gasterosteiformes		 管口鱼科
Aulostomidae		 管口鱼属
Aulostomus		 中华管口鱼
A. chinensis		 A 0.00 0.00000 0.13 0.00031
鲈形目
Perciformes		 刺尾鱼科
Acanthuridae		 鼻鱼属
Naso		 六棘鼻鱼
N. hexacanthus		 A 0.97 0.00215 0.00 0.00000
突角鼻鱼
N. annulatus		 A 0.31 0.00068 0.28 0.00071
高鳍刺尾鱼属
Zebrasoma		 黄高鳍刺尾鱼
Z. flavescens		 A 0.04 0.00005 0.00 0.00000
带鱼科
Trichiuridae		 带鱼属
Trichiurus		 带鱼属未定种
Trichiurus sp.		 C 0.00 0.00000 1.02 0.00256
笛鲷科
Lutjanidae		 笛鲷属
Lutjanus		 四带笛鲷
L. kasmira		 A 0.38 0.00043 0.90 0.00224
蝴蝶鱼科
Chaetodontidae		 蝴蝶鱼属
Chaetodon		 叉纹蝴蝶鱼
C. auripes		 A 0.07 0.00015 0.00 0.00000
隆头鱼科
Labridae		 唇鱼属
Cheilinus		 横带唇鱼
C. fasciatus		 A 0.02 0.00002 0.00 0.00000
海猪鱼属
Halichoeres		 三斑海猪鱼
H. trimaculatus		 A 6.66 0.03700 0.08 0.00039
裸颊鲷科
Lethrinidae		 裸颊鲷属
Lethrinus		 桔带裸颊鲷
L. obsoletus		 A 0.02 0.00003 0.00 0.00000
梅鲷科
Caesionidae		 鳞鳍梅鲷属
Pterocaesio		 黑带鳞鳍梅鲷
P. tile		 A 0.00 0.00000 6.32 0.01581
鮨科
Serranidae		 侧牙鲈属
Variola		 侧牙鲈
V. louti		 A 0.06 0.00007 0.00 0.00000
鲭科Scombridae 舵鲣属
Auxis		 扁舵鲣
A. thazard		 B 0.01 0.00001 0.00 0.00000
鲣属
Katsuwonus		 鲣
K. pelamis		 B 22.35 0.09931 0.00 0.00000
金枪鱼属
Thunnus		 金枪鱼属未定种
Thunnus sp.		 B 0.18 0.00020 0.00 0.00000
鲔属
Euthynnus		 鲔
E. affinis		 B 0.04 0.00005 0.00 0.00000
鲹科
Carangidae		 鲹属
Caranx		 珍鲹
C. ignobilis		 A 0.54 0.00120 0.00 0.00000
圆鲹属
Decapterus		 圆鲹属未定种
Decapterus sp.		 B 65.44 0.50900 0.35 0.00087
双边鱼科
Ambassidae		 双边鱼属
Ambassis		 双边鱼属未定种
Ambassis sp.		 B 0.00 0.00000 0.09 0.00024
羊鱼科
Mullidae		 副绯鲤属
Parupeneus		 多带副绯鲤
P. multifasciatus		 A 0.03 0.00003 8.49 0.04247
拟羊鱼属
Mulloidichthys		 无斑拟羊鱼
M. vanicolensis		 A 0.30 0.00034 0.00 0.00000
鹦嘴鱼科
Scaridae		 大鹦嘴鱼属
Bolbometopon		 隆头大鹦嘴鱼
B. muricatum		 A 0.37 0.00041 0.06 0.00016
绿鹦嘴鱼属
Chlorurus		 灰鹦嘴鱼
C. sordidus		 A 0.38 0.00085 16.94 0.08470
鹦嘴鱼属
Scarus		 绿唇鹦嘴鱼
S. forsteni		 A 1.35 0.00300 60.24 0.30120
许氏鹦嘴鱼
S. schlegeli		 A 0.01 0.00001 3.92 0.00979
鼠鱚目
Gonorhynchiformes		 遮目鱼科
Chanidae		 遮目鱼属
Chanos		 遮目鱼
C. chanos		 C 0.45 0.00050 0.00 0.00000
鲀形目
Tetraodontiformes		 单角鲀科
Monacanthidae		 前角鲀属
Pervagor		 前角鲀属未定种
Pervagor sp.		 A 0.00 0.00000 1.18 0.00295

注: A. 礁栖鱼类; B. 大洋性中上层鱼类; C. 深海鱼类

从优势度结果看, 永乐环礁和东岛鱼卵优势种组成差异明显。永乐环礁的优势种是圆鲹属未定种(Decapterus sp., 相对丰度65.44%, 优势度0.50900)、鲣(Katsuwonus pelamis, 相对丰度22.35%, 优势度0.09931)和三斑海猪鱼(Halichoeres trimaculatus, 相对丰度6.66%, 优势度0.03700); 东岛的优势种是绿唇鹦嘴鱼(Scarus forsteni, 相对丰度60.24%, 优势度0.30120)、灰鹦嘴鱼(Chlorurus sordidus, 相对丰度16.94%, 优势度0.08470)和多带副绯鲤(Parupeneus multifasciatus, 相对丰度8.49%, 优势度0.04247)(表1)。
从相似度来看, 永乐环礁和东岛之间的相似性指数均为不相似, 其中目、科、属、种的相似度指数分别是25.00%、37.50%、32.00%和33.33%。

2.3 生态类型和生物多样性

在永乐环礁和东岛产卵的鱼类三种生态类型均有分布, 种类组成上均以礁栖鱼类占据优势, 永乐环礁中占72.73%(16种), 东岛中占78.57%(11种); 其次为大洋性中上层鱼类, 永乐环礁中占22.73%(5种), 东岛中占14.29%(2种); 深海鱼类最少, 永乐环礁中占4.55%(1种), 东岛中占7.14%(1种)(图3)。鱼卵相对丰度上, 永乐环礁以大洋性中上层鱼类(88.02%)最多, 其次为礁栖鱼类(11.53%), 深海鱼类(0.45%)最少; 而东岛以礁栖鱼类(98.54%)最多, 大洋性中上层鱼类(0.44%)和深海鱼类(1.02%)均较低 (图4)。永乐环礁的物种丰富度指数(D)为2.46, 明显高于东岛的1.87, 而香农-威纳多样性指数和均匀度指数均表现为永乐环礁(1.08和0.35), 低于东岛 (1.32和0.50)(图5)。
图3 永乐环礁和东岛鱼卵不同生态类型的种类组成

Fig. 3 Species composition of different ecological types of fish eggs in the Yongle atoll and Dongdao Island

图4 永乐环礁和东岛鱼卵不同生态类型的相对丰度

Fig. 4 Relative abundance of different ecological types of fish eggs in the Yongle atoll and Dongdao Island

图5 永乐环礁和东岛的鱼卵生物多样性指数

Fig. 5 Diversity indexes of parrotfish in the Yongle atoll and Dongdao Island

3 讨论

3.1 鱼卵种类组成

此次调查共获得27种鱼卵, 种类符合西沙群岛鱼类群落结构特征(李永振 等, 2007; 邱书婷 等, 2022; 王腾 等, 2022b)。与其他岛礁鱼卵组成相似, 均以鲈形目占据优势, 例如南沙美济礁(91.89%)(卢芷程 等, 2021)、南沙永暑礁(100%) (吴娜 等, 2018)、西沙七连屿(91.78%)(闫亚利, 2019)和肯尼亚潟湖(91%)(Okemwa et al, 2019)。本研究发现, 两区域鱼卵在种类组成上均以礁栖鱼类为主, 同时也发现了一定比例的大洋性中上层鱼类和深海鱼类; 而在鱼卵相对丰度上, 永乐环礁以大洋性鱼类为主, 东岛以礁栖鱼类为主。这体现了珊瑚岛礁作为海洋鱼类产卵场的功能, 在主要支持了礁栖鱼类鱼卵的同时, 还为大洋性中上层鱼类和深海鱼类提供了重要的繁殖场所(Pinheiro et al, 2021)。
大洋性中上层鱼类中主要为金枪鱼类, 本次研究在永乐环礁发现4种金枪鱼的鱼卵, 且拥有较高的序列丰度, 而东岛仅发现其中一种, 且丰度较低。说明金枪鱼类选择了永乐环礁作为主要产卵场, 而东岛的金枪鱼鱼卵有可能为漂流所致。这可能与永乐环礁拥有开放的潟湖有关, 开放性的潟湖不仅可以为金枪鱼的繁殖提供稳定的环境因子, 而且其中的洋流和旋涡也是金枪鱼幼鱼摄食、运输和生长的重要机制(Muhling et al, 2017; 卢芷程 等, 2021; Schiller et al, 2021)。 南海水产研究所分别于1964年3月—1965年2月、1975年1月—1976年5月和1978年1月—1979年2月三次大规模对西中沙群岛海域的金枪鱼类鱼卵仔鱼进行了调查, 均采集到黄鳍金枪鱼、鲣和扁舵鲣的仔鱼(张仁斋, 1983)。闫亚利(2019)在2017年冬季和2018年春、秋季调查了西沙七连屿的鱼卵仔鱼, 采集到了鲭科的圆鮀鲣(Auxis rochei)和扁鮀鲣的鱼卵。结合上述研究, 均说明西沙群岛为金枪鱼的产卵场, 而本研究进一步为永乐环礁作为金枪鱼产卵场之一提供了直接证据。
另外, 本次调查鉴定出了双边鱼(Ambassis sp.)这一未记录种, 对比以往西沙研究(孙典荣 等, 2005; Qiu et al, 2021; 王腾 等, 2022a; 王腾 等, 2022b), 均未发现相关记录。卢芷程 等(2021)使用分子方法在对南沙美济礁鱼卵仔鱼的调查中也发现了捕捞调查中未记录的圆翅燕鱼(Platax pinnatus)、颌圆鲹(Decapterus macarellus)和环状钝头鱼(Cymolutes torquatus)等未记录种。de Lima等(2020)用分子方法调查了巴拉那帕内马河的鱼卵仔鱼分布, 通过对鱼卵DNA条形码的鉴定, 发现了鲇形目的两种稀有鱼类芒拟油鲇(Pseudopimelodus mangurus)和闪光鸭嘴鲇(Pseudoplatystoma corruscans)。这些均显示了基于高通量测序的方法对于未记录种有更好的发现能力, 可作为传统鱼类资源调查的有效补充方法(Elias-Gutierrez et al, 2021)。

3.2 鱼卵资源量和生物多样性特征比较

本次调查永乐环礁和东岛的每个采样点均出现了鱼卵, 并且密度较高。一般认为温度、盐度、溶解氧、pH和栖息地特征等环境因子共同影响鱼卵仔鱼分布(Song et al, 2019)。西沙群岛属于热带海洋季风气候, 具有高温低盐的特征, 从三月到十一月温度就保持在28℃左右, 其中四月到六月是鱼类繁殖盛期(陈真然 等, 1978; 陈真然, 1979)。因此, 调查期间鱼卵表现出较高的密度和出现率, 并进一步证明了珊瑚礁生态系统对于海洋鱼类繁殖的重要性(Siddon et al, 2019; Zhang et al, 2022)。
永乐环礁鱼卵资源量明显高于东岛。可能是永乐环礁中间有开放性的潟湖, 与东岛相比拥有更复杂的珊瑚礁结构、更多的产卵微生境、更大的礁外运输率和更可靠的食物获得性, 更适宜作为鱼类的产卵场和育幼场(Gladstone, 2007)。并且, 与物理上相对封闭的潟湖相比, 开放性的潟湖对于鱼类的繁殖更具优势, 尤其是大洋性鱼类一般不会选择半封闭的水体, 那会阻止远洋繁殖鱼类和仔稚鱼离开出生地(Leis et al, 2003)。Vásquez-Yeomans等(2003)调查了墨西哥新克洛浅滩的鱼卵仔鱼, 发现与海洋区相比潟湖中的鱼卵仔鱼丰度更高。且开放性的潟湖更符合鱼卵仔鱼的漂移模式, 其中的洋流促进了鱼卵仔鱼广泛的扩散, 增加了仔鱼找到另一个栖息地的可能(Gladstone, 2007)。Machado等(2021)评估了间歇性封闭-开放潟湖与邻近水域的连通性, 发现了两种环境之间鱼卵仔鱼交换的明显证据, 并且这种连通性的对浮游鱼类的存活率和遗传流动有潜在的贡献。因此, 开放性的潟湖对鱼类的繁殖活动具有重要的意义。
永乐环礁鱼卵物种丰富度指数明显高于东岛, 但由于单一优势种的大量出现, 香农-威纳多样性指数和均匀度指数均低于东岛, 表明永乐环礁较东岛拥有更丰富的产卵群体, 但是存在少数物种占据了大部分产卵资源的现象, 这是许多产卵场的共同特征(Pinheiro et al, 2009; Farhana-Azmi et al, 2022)。Saroj等(2020)于2017—2019年在马纳尔湾水域的曼达帕姆、图图库迪和蓬纳卡亚尔三个站点对鱼卵仔鱼的多样性和分布进行了研究, 发现三个地点均在不同时间出现了少数物种占据大部分资源的情况, 导致同一地点在某一时期鱼卵仔鱼物种丰富度指数较高, 而多样性指数和均匀度指数较低。杭州湾也有这样的特点, 由于髭缟虾虎鱼(Tridentiger barbatus)作为优势种的出现, 使得杭州湾北部水域仔稚鱼在5、6月份与4月份相比物种丰富度指数很高, 但Shannon-Wiener指数和Pielou均匀度指数均很低(闫欣 等, 2014)。徐华 等(2021)调查了调水调沙期间黄河口及邻近海域鱼卵仔稚鱼的群落结构动态变化, 发现2011年调水调沙前的鱼卵仔鱼物种丰富度明显高于调水调沙后, 而多样性指数和均匀度指数则相反。这种现象可能与鱼类产卵场分布和繁殖策略相关, 可以降低鱼卵仔鱼的种间竞争, 更有利于鱼类种群的繁衍壮大, 但具体关系还需进一步研究。
本文通过高通量的方法对不同采样点的混合样品进行扩增和测序, 与传统单一物种鉴定方法相比, 高通量测序方法能够在短时间内低成本的对大量复合样品进行物种鉴定。但是也存在局限性, 比如不能将鱼卵样品与鉴定种类对应, 不能精确确定不同种类鱼卵的丰度等, 这个问题可以作为今后的研究方向。

4 结论

这项研究证明了高通量测序技术在鱼卵鉴定方面的有效性, 可以作为渔业资源调查的补充手段。另外, 分析了永乐环礁和东岛鱼卵的种类组成和分布特征, 二者可能均为各种海洋鱼类的产卵场, 并进一步探讨了永乐环礁的开放性潟湖对于海洋鱼类, 尤其是大洋性鱼类繁殖活动的重要性。

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