基于线粒体CO#cod#x02160;基因对南海毛颚动物箭虫科的鉴定及系统发育初探

doi:10.11978/2017043

热带海洋学报 ›› 2017, Vol. 36 ›› Issue (6): 100-108.doi: 10.11978/2017043CSTR: 32234.14.2017043

所属专题：南海专题

基于线粒体CO#cod#x02160;基因对南海毛颚动物箭虫科的鉴定及系统发育初探

陈斌(), 何薇, 李海涛, 吴鹏, 肖瑜璋, 吕向立(), 何静

国家海洋局南海环境监测中心, 广东广州 510300

收稿日期:2017-04-11 修回日期:2017-06-08 出版日期:2017-11-30 发布日期:2018-01-18
作者简介:
作者简介：陈斌(1988#cod#x02014;), 女, 助理工程师, 生物工程硕士, 主要从事海洋环境监测。E-mail: stlxbb@126.com
基金资助:
广东省自然科学基金项目(2014A030310495);国家海洋局南海分局海洋科学技术局长基金项目(1513)

Identification and phylogenetic analysis of Sagittidae (Chaetognatha) in the South China Sea based on mitochondrial CO#cod#x02160; gene

Bin CHEN(), Wei HE, Haitao LI, Peng WU, Yuzhang Xiao, Xiangli LYU(), Jing HE

South China Sea Environmental Monitoring Center, State Oceanic Administration, People#cod#x02019;s Republic of China, Guangzhou 510300, China

Received:2017-04-11 Revised:2017-06-08 Online:2017-11-30 Published:2018-01-18
About author:
Author:QIU Chunhua.E-mail: qiuchh3@mail. sysu.edu.cn
Supported by:
Guangdong Natural Science Foundation (2014A030310495);Director Fund of Marine Science and Technology, South China Sea Branch, State Oceanic Administration, People#cod#x02019;s Republic of China (1513)

摘要/Abstract

摘要：

为提高毛颚动物鉴定的准确性, 进一步探索毛颚动物箭虫纲箭虫科各属的系统发育地位, 文章采用形态学鉴定与DNA条形码技术相结合, 对南海北部采集的部分箭虫种类进行鉴定, 并初步探讨其系统分类地位。结果表明包括粗壮猛箭虫(Ferosagitta robusta)、小型滨箭虫(Aidanosagitta neglecta)、规则滨箭虫(Aidanosagitta regularis)在内的箭虫, 其形态鉴定结果与分子鉴定的结果一致, 其CO#cod#x02160;(线粒体编码的细胞色素氧化酶第一亚基, cytochrome oxidase subunit #cod#x02160;)序列在系统发育树中与GenBank得到的相应箭虫CO#cod#x02160;序列以91%~100%高支持率形成各自单系枝。种内遗传距离范围为0.000~0.015, 平均值为0.004; 种间遗传距离范围为0.081~0.173, 平均值为0.127; 种内遗传距离和种间遗传距离未重叠, 存在明显的条形码间隙, 证实了CO#cod#x02160;基因作为毛颚动物DNA条形码标准基因的可行性。本研究首次将DNA条形码技术应用于箭虫科内各属的系统发育分析, 首次从分子层面探讨其系统发育地位。

关键词: DNA条形码, CO#cod#x02160;, 基因, 系统发育, 毛颚动物, 箭虫科

Abstract:

To improve the accuracy of Chaetognatha identification, we combined morphological identification and DNA barcoding based on the mitochondrial CO#cod#x02160;(cytochrome oxidase subunit #cod#x02160;) gene to identify some arrow worms from the northern South China Sea. The results showed that the morphologically and molecularly identified results were consistent, for the species of Ferosagitta robusta, Aidanosagitta neglecta and Aidanosagitta regularis. Furthermore, the neighbor-joining phylogenetic tree was constructed with the amplified CO#cod#x02160; sequences and those homologous sequences from GenBank based on Kimura 2-parameter, and the phylogenetic positions of Sagiitta (Sagittidae, Sagittoidea) were discussed. The results indicated that these CO#cod#x02160; sequences formed monophylotic clades compared with GenBank (91%~100%). The intraspecific genetic distance ranged from 0.000 to 0.015 with an average value of 0.004, while the interspecific genetic distance ranged from 0.081 to 0.173 with an average value of 0.127. The barcoding gap was detected between intra and interspecific variations, suggesting that the CO#cod#x02160; gene could be a standard DNA barcode for the Chaetognatha identification. This study was the first to apply DNA barcoding to phylogenetic analysis of Sagittidae, and to discuss the phylogenetic position at molecular level.

Key words: DNA barcoding, CO#cod#x02160;, gene, phylogeny, Chaetognatha, Sagittidae

中图分类号:

Q178.53

陈斌, 何薇, 李海涛, 吴鹏, 肖瑜璋, 吕向立, 何静. 基于线粒体CO#cod#x02160;基因对南海毛颚动物箭虫科的鉴定及系统发育初探[J]. 热带海洋学报, 2017, 36(6): 100-108.

Bin CHEN, Wei HE, Haitao LI, Peng WU, Yuzhang Xiao, Xiangli LYU, Jing HE. Identification and phylogenetic analysis of Sagittidae (Chaetognatha) in the South China Sea based on mitochondrial CO#cod#x02160; gene[J]. Journal of Tropical Oceanography, 2017, 36(6): 100-108.

图/表 6

表1

图 1

图 2

表2

表 3

基于CO#cod#x02160;基因序列通过K2P模型计算的箭虫科遗传距离"

序号	种名	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19
1	Ferosagitta robusta
2	Ferosagitta robusta	0.006
3	Ferosagitta robusta	0.002	0.004
4	Ferosagitta robusta	0.002	0.004	0.000
5	Ferosagitta ferox	0.083	0.086	0.081	0.081
6	Ferosagitta ferox	0.083	0.086	0.081	0.081	0.000
7	Ferosagitta ferox	0.083	0.086	0.081	0.081	0.000	0.000
8	Aidanosagitta regularis	0.141	0.143	0.143	0.143	0.160	0.160	0.160
9	Aidanosagitta regularis	0.141	0.143	0.143	0.143	0.160	0.160	0.160	0.004
10	Aidanosagitta neglecta	0.131	0.129	0.129	0.129	0.121	0.121	0.121	0.138	0.143
11	Aidanosagitta neglecta	0.131	0.129	0.129	0.129	0.121	0.121	0.121	0.138	0.143	0.000
12	Aidanosagitta neglecta	0.129	0.126	0.126	0.126	0.124	0.124	0.124	0.141	0.145	0.006	0.006
13	Aidanosagitta crassa	0.131	0.129	0.133	0.133	0.099	0.099	0.099	0.138	0.143	0.124	0.124	0.126
14	Sagitta spp.	0.092	0.090	0.094	0.094	0.105	0.105	0.105	0.141	0.141	0.136	0.136	0.138	0.103
15	Sagitta spp.	0.096	0.094	0.099	0.099	0.096	0.096	0.096	0.141	0.141	0.126	0.126	0.129	0.103	0.015
16	Sagitta spp.	0.092	0.090	0.094	0.094	0.105	0.105	0.105	0.141	0.141	0.136	0.136	0.138	0.103	0.000	0.015
17	Zonosagitta nagae	0.136	0.143	0.138	0.138	0.112	0.112	0.112	0.124	0.129	0.119	0.119	0.126	0.092	0.121	0.121	0.121
18	Zonosagitta bedoti	0.143	0.150	0.145	0.145	0.115	0.115	0.115	0.155	0.155	0.126	0.126	0.129	0.112	0.143	0.138	0.143	0.121
19	Sagitta bipunctata	0.129	0.126	0.126	0.126	0.143	0.143	0.143	0.121	0.126	0.141	0.141	0.138	0.126	0.148	0.143	0.148	0.145	0.143
20	Flaccisagitta enflata	0.143	0.150	0.145	0.145	0.173	0.173	0.173	0.170	0.170	0.150	0.150	0.153	0.160	0.148	0.153	0.148	0.160	0.168	0.153

表 3

图 3

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种名	数量	主要形态特征
粗壮猛箭虫 Ferosagitta robusta	15	体长10~12mm, 体坚硬, 不透明, 身体从头到尾隔的宽度相等。颈部明显, 领发达, 肠分歧, 卵巢宽大, 卵呈梨形, 储精囊呈倒梨型, 前鳍椭圆形, 后鳍呈圆形, 尾鳍呈三角形
肥胖软箭虫 Flaccisagitta enflata	5	体长8~20mm, 体肥胖、柔软、透明, 颈部短, 无领, 肠不分歧, 卵巢短, 卵呈圆形, 储精囊呈球形, 前鳍仅边缘具鳍条, 尾鳍呈半圆形
凶型猛箭虫 Ferosagitta ferox	8	体长9~16mm, 体坚实、不透明, 头大, 颈部明显, 肠分歧, 卵巢呈管状, 向前伸展可达颈部, 储精囊呈长卵圆形, 后鳍中央具一条狭小无鳍条带
规则滨箭虫 Aidanosagitta regularis	6	体长6mm, 体细长, 较坚硬, 颈部不明显, 头到前鳍前半部的领非常发达, 其余为薄层延伸到尾, 卵巢前伸达腹神经节, 储精囊呈耳形
小型滨箭虫 Aidanosagitta neglecta	6	体长6~9mm, 体坚硬、不透明, 颈部不明显, 领从颈到前鳍前端厚, 肠分歧, 卵巢充满体腔, 储精囊呈卵圆形, 接近后鳍但与尾鳍分离, 前后鳍均充满鳍条
未定种 Sagitta spp.	3	体长15~16mm, 体坚实、不透明, 头大, 颈部明显。卵巢达前鳍中部, 卵大而圆, 储精囊呈耳状, 前鳍较后鳍长且大, 后鳍稍圆, 其中央无鳍条

基于线粒体CO#cod#x02160;基因对南海毛颚动物箭虫科的鉴定及系统发育初探

Identification and phylogenetic analysis of Sagittidae (Chaetognatha) in the South China Sea based on mitochondrial CO#cod#x02160; gene

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序号	分类地位	中文种名	拉丁种名	GenBank登录号	来源
1	箭虫科猛箭虫属	粗壮猛箭虫	Ferosagitta robusta	KY882128	南海北部
2	箭虫科猛箭虫属	粗壮猛箭虫	Ferosagitta robusta	KY882126	南海北部
3	箭虫科猛箭虫属	粗壮猛箭虫	Ferosagitta robusta	KY882127	南海北部
4	箭虫科猛箭虫属	凶型猛箭虫	Ferosagitta ferox	KY882123	南海北部
5	箭虫科猛箭虫属	凶型猛箭虫	Ferosagitta ferox	KY882124	南海北部
6	箭虫科猛箭虫属	凶型猛箭虫	Ferosagitta ferox	KY882125	南海北部
7	箭虫科猛箭虫属	规则滨箭虫	Aidanosagitta regularis	KY882129	南海北部
8	未定	未定种	Sagitta spp.	KY882132	南海北部
9	未定	未定种	Sagitta spp.	KY882133	南海北部
10	未定	未定种	Sagitta spp.	KY882134	南海北部
11	箭虫科滨箭虫属	小型滨箭虫	Aidanosagitta neglecta	KY882130	南海北部
12	箭虫科滨箭虫属	小型滨箭虫	Aidanosagitta neglecta	KY882131	南海北部
13	箭虫科猛箭虫属	粗壮猛箭虫	Ferosagitta robusta	JN258035.1	GenBank
14	箭虫科滨箭虫属	正形滨箭虫	Aidanosagitta regularis	FJ538304.1	GenBank
15	箭虫科软箭虫属	肥胖软箭虫	Flaccisagitta enflata	KF977332	GenBank
16	箭虫科滨箭虫属	强壮滨箭虫	Aidanosagitta crassa	HQ700947.1	GenBank
17	箭虫科带箭虫属	纳嘎带箭虫	Zonosagitta nagae	NC013810.1	GenBank
18	箭虫科带箭虫属	百陶带箭虫	Zonosagitta bedoti	JN258005.1	GenBank
19	箭虫科滨箭虫属	小型滨箭虫	Aidanosagitta neglecta	JN258022	GenBank
20	箭虫科箭虫属	双斑箭虫	Sagitta bipunctata	GQ368396	GenBank

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