生物多样性 ›› 2017, Vol. 25 ›› Issue (8): 847-855. DOI: 10.17520/biods.2017094
收稿日期:
2017-03-23
接受日期:
2017-06-19
出版日期:
2017-08-20
发布日期:
2017-08-31
通讯作者:
张秀梅
作者简介:
# 共同第一作者 Co-first authors
基金资助:
Xiaomeng Zhou1, Shuxin Guo1, Na Song1, Xiumei Zhang1,2,*()
Received:
2017-03-23
Accepted:
2017-06-19
Online:
2017-08-20
Published:
2017-08-31
Contact:
Zhang Xiumei
摘要:
为了准确鉴定2015年7-10月青岛崂山青山湾鱼类浮游生物调查中采集的依形态学初步鉴定为某种舌鳎属的鱼卵、仔鱼样本, 本实验同时采集青岛胶州湾短吻红舌鳎(Cynoglossus joyneri)和长吻红舌鳎(C. lighti)的成鱼样本为参照, 通过线粒体COI基因测序分析, 以及同山东半岛海域分布的6种舌鳎属鱼类的COI基因序列比对, 开展了该舌鳎属鱼卵、仔鱼种类的鉴别分析, 以期为今后舌鳎属鱼类的分类及其鱼卵、仔鱼的鉴别提供参考资料。形态学研究结果显示: 鱼卵为圆球形浮性卵, 直径0.68-0.87 mm (0.73 ± 0.03 mm, n = 50), 卵膜单层, 较薄、光滑透明, 卵周隙较大, 卵膜及卵黄囊均无特殊构造, 油球6-15个, 直径0.04-0.10 mm (0.07 ± 0.01 mm, n = 50), 圆形, 大小不一, 多位于卵黄囊中与胚体相对的一侧, 既有聚集分布也有分散分布; 仔鱼的2条冠状幼鳍及右眼移位过程中体长、色素分布等的特征和变化是此鱼种仔鱼鉴别的最明显特征。遗传学分析结果显示: 该种舌鳎属样品(Cynoglossus sp.)与短吻红舌鳎和长吻红舌鳎的遗传距离最为接近, 三者K2P遗传距离为0.006-0.009; 短吻三线舌鳎(C. abbreviatus)与紫斑舌鳎(C. purpureomaculatus)亲缘关系最近, 两者K2P遗传距离仅为0.002。以带纹条鳎(Zebrias zebra)为外类群构建的NJ系统发育树显示, 该种舌鳎属样品与短吻红舌鳎和长吻红舌鳎以及短吻三线舌鳎与紫斑舌鳎分别聚为一支, 其他种类各聚为一支。综合形态特征和COI基因比对结果, 确定该舌鳎属鱼卵、仔鱼样品为短吻红舌鳎。
周晓梦, 郭书新, 宋娜, 张秀梅 (2017) 短吻红舌鳎鱼卵、仔鱼的DNA条形码和形态学鉴定. 生物多样性, 25, 847-855. DOI: 10.17520/biods.2017094.
Xiaomeng Zhou, Shuxin Guo, Na Song, Xiumei Zhang (2017) Identification of Cynoglossus joyneri eggs and larvae by DNA barcoding and morphological method. Biodiversity Science, 25, 847-855. DOI: 10.17520/biods.2017094.
形态特征 Morphological characters | 短吻红舌鳎 C. joyneri | 长吻红舌鳎 C. lighti | ||||
---|---|---|---|---|---|---|
Li & Wang, 1995 | Zhou & Cheng, 1997 | Nakabo, 2000 | Li & Wang, 1995 | Zhou & Cheng, 1997 | Nakabo, 2000 | |
上、中侧线鳞间鳞数 Scales between upper and middle lateral line | 12-13 | 11-12 | / | 10-11 | 10-11 | / |
体长/体高 Body length/body depth | 3.6-4.4 | 3.8-4.0 | 3.4-4.1 | 3.8-4.5 | 4.2-5.0 | 3.8-4.5 |
头长/头高 Head length/head depth | ≤1 | / | / | >1 | / | / |
表1 中外相关文献中短吻红舌鳎和长吻红舌鳎的形态学数据记录的比较
Table 1 Comparative counts and distribution of Cynoglossus joyneri and C. lighti in different records
形态特征 Morphological characters | 短吻红舌鳎 C. joyneri | 长吻红舌鳎 C. lighti | ||||
---|---|---|---|---|---|---|
Li & Wang, 1995 | Zhou & Cheng, 1997 | Nakabo, 2000 | Li & Wang, 1995 | Zhou & Cheng, 1997 | Nakabo, 2000 | |
上、中侧线鳞间鳞数 Scales between upper and middle lateral line | 12-13 | 11-12 | / | 10-11 | 10-11 | / |
体长/体高 Body length/body depth | 3.6-4.4 | 3.8-4.0 | 3.4-4.1 | 3.8-4.5 | 4.2-5.0 | 3.8-4.5 |
头长/头高 Head length/head depth | ≤1 | / | / | >1 | / | / |
编号Number | 种类 Species | 序列编号Sequence numbers | 序列来源 Sources of sequences |
---|---|---|---|
1 | 舌鳎属样品 Cynoglossus sp. | egg.1-egg.3; lar.1-lar.3 | 本研究 This study |
2 | 短吻红舌鳎 C. joyneri | joy.1-joy.5 | 本研究(1-3); JQ738430.1; KF979127.1 |
3 | 长吻红舌鳎 C. lighti | lig.1-lig4 | 本研究(1-2); DQ116752.1; HQ711865.1 |
4 | 宽体舌鳎 C. robustus | rob.1-rob.3 | KT718591.1; HM180552.1; HM180553.1 |
5 | 半滑舌鳎 C. semilaevis | sem.1-sem.3 | JN242742.1; JN242743.1; JN242744.1 |
6 | 紫斑舌鳎 C. purpureomaculatus | pur.1-pur.3 | DQ116749.1; JQ738570.1; JQ738571.1 |
7 | 短吻三线舌鳎 C. abbreviatus | abb.1 | DQ116750.1 |
8 | 带纹条鳎 Zebrias zebra | zeb.1-zeb.3 | JQ738566.1; JQ738567.1; JQ738568.1 |
表2 样品与COI基因序列信息
Table 2 Information of Cynoglossus samples and COI gene sequences
编号Number | 种类 Species | 序列编号Sequence numbers | 序列来源 Sources of sequences |
---|---|---|---|
1 | 舌鳎属样品 Cynoglossus sp. | egg.1-egg.3; lar.1-lar.3 | 本研究 This study |
2 | 短吻红舌鳎 C. joyneri | joy.1-joy.5 | 本研究(1-3); JQ738430.1; KF979127.1 |
3 | 长吻红舌鳎 C. lighti | lig.1-lig4 | 本研究(1-2); DQ116752.1; HQ711865.1 |
4 | 宽体舌鳎 C. robustus | rob.1-rob.3 | KT718591.1; HM180552.1; HM180553.1 |
5 | 半滑舌鳎 C. semilaevis | sem.1-sem.3 | JN242742.1; JN242743.1; JN242744.1 |
6 | 紫斑舌鳎 C. purpureomaculatus | pur.1-pur.3 | DQ116749.1; JQ738570.1; JQ738571.1 |
7 | 短吻三线舌鳎 C. abbreviatus | abb.1 | DQ116750.1 |
8 | 带纹条鳎 Zebrias zebra | zeb.1-zeb.3 | JQ738566.1; JQ738567.1; JQ738568.1 |
碱基 Base | 总频率 Total | 密码子第1位点 The first position of codon | 密码子第2位点 The second position of codon | 密码子第3位点 The third position of codon |
---|---|---|---|---|
T | 31.3 | 18.0 | 43.8 | 32.1 |
C | 26.5 | 24.5 | 28.1 | 27.1 |
A | 25.6 | 27.4 | 14.5 | 34.8 |
G | 16.6 | 30.1 | 13.6 | 6.0 |
A+T | 56.9 | 45.4 | 58.3 | 66.9 |
G+C | 43.1 | 54.6 | 41.7 | 33.1 |
表3 COI基因部分序列中各碱基平均分布概率(%)
Table 3 Average nucleotide frequencies (%) of COI partial sequence of eight species
碱基 Base | 总频率 Total | 密码子第1位点 The first position of codon | 密码子第2位点 The second position of codon | 密码子第3位点 The third position of codon |
---|---|---|---|---|
T | 31.3 | 18.0 | 43.8 | 32.1 |
C | 26.5 | 24.5 | 28.1 | 27.1 |
A | 25.6 | 27.4 | 14.5 | 34.8 |
G | 16.6 | 30.1 | 13.6 | 6.0 |
A+T | 56.9 | 45.4 | 58.3 | 66.9 |
G+C | 43.1 | 54.6 | 41.7 | 33.1 |
种类 Species | 样本量 Sample size | 平均GC含量 Average GC content (%) | |||
---|---|---|---|---|---|
全部位点 Total positions | 第1位点 The first position | 第2位点 The second position | 第3位点 The third position | ||
1舌鳎属样品 Cynoglossus sp. | 6 | 42.9 | 57.2 | 42 | 29.5 |
2短吻红舌鳎 C. joyneri | 5 | 43 | 56.9 | 42 | 29.9 |
3长吻红舌鳎 C. lighti | 4 | 43 | 57.3 | 42 | 29.9 |
4宽体舌鳎 C. robustus | 3 | 42.5 | 53.3 | 42 | 31.9 |
5半滑舌鳎 C. semilaevis | 3 | 40.8 | 52.4 | 42 | 28.2 |
6紫斑舌鳎 C. purpureomaculatus | 3 | 43.1 | 52.3 | 42 | 34.9 |
7短吻三线舌鳎 C. abbreviatus | 1 | 42.9 | 51.8 | 42 | 34.9 |
8带纹条鳎 Zebrias zebra | 3 | 48.4 | 53.9 | 42.5 | 49.0 |
表4 8种鱼类COI基因密码子位点GC含量
Table 4 The GC content of COI partial sequence for eight species in all codon positions
种类 Species | 样本量 Sample size | 平均GC含量 Average GC content (%) | |||
---|---|---|---|---|---|
全部位点 Total positions | 第1位点 The first position | 第2位点 The second position | 第3位点 The third position | ||
1舌鳎属样品 Cynoglossus sp. | 6 | 42.9 | 57.2 | 42 | 29.5 |
2短吻红舌鳎 C. joyneri | 5 | 43 | 56.9 | 42 | 29.9 |
3长吻红舌鳎 C. lighti | 4 | 43 | 57.3 | 42 | 29.9 |
4宽体舌鳎 C. robustus | 3 | 42.5 | 53.3 | 42 | 31.9 |
5半滑舌鳎 C. semilaevis | 3 | 40.8 | 52.4 | 42 | 28.2 |
6紫斑舌鳎 C. purpureomaculatus | 3 | 43.1 | 52.3 | 42 | 34.9 |
7短吻三线舌鳎 C. abbreviatus | 1 | 42.9 | 51.8 | 42 | 34.9 |
8带纹条鳎 Zebrias zebra | 3 | 48.4 | 53.9 | 42.5 | 49.0 |
种间 Pairwise-species | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
---|---|---|---|---|---|---|---|---|
1舌鳎属样品 Cynoglossus sp. | 0.009 | |||||||
2短吻红舌鳎 C. joyneri | 0.009 | 0.008 | ||||||
3长吻红舌鳎 C. lighti | 0.008 | 0.006 | 0.004 | |||||
4宽体舌鳎 C. robustus | 0.236 | 0.240 | 0.240 | 0.002 | ||||
5半滑舌鳎 C. semilaevis | 0.189 | 0.192 | 0.190 | 0.208 | 0.000 | |||
6紫斑舌鳎 C. purpureomaculatus | 0.181 | 0.182 | 0.181 | 0.200 | 0.113 | 0.000 | ||
7短吻三线舌鳎C. abbreviatus | 0.182 | 0.185 | 0.184 | 0.203 | 0.116 | 0.002 | n/c | |
8带纹条鳎 Zebrias zebra | 0.245 | 0.245 | 0.244 | 0.277 | 0.278 | 0.302 | 0.304 | 0.002 |
表5 8种鱼类的种间遗传距离和种内遗传距离
Table 5 Genetic distance pairwise-species and within-species of eight species
种间 Pairwise-species | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
---|---|---|---|---|---|---|---|---|
1舌鳎属样品 Cynoglossus sp. | 0.009 | |||||||
2短吻红舌鳎 C. joyneri | 0.009 | 0.008 | ||||||
3长吻红舌鳎 C. lighti | 0.008 | 0.006 | 0.004 | |||||
4宽体舌鳎 C. robustus | 0.236 | 0.240 | 0.240 | 0.002 | ||||
5半滑舌鳎 C. semilaevis | 0.189 | 0.192 | 0.190 | 0.208 | 0.000 | |||
6紫斑舌鳎 C. purpureomaculatus | 0.181 | 0.182 | 0.181 | 0.200 | 0.113 | 0.000 | ||
7短吻三线舌鳎C. abbreviatus | 0.182 | 0.185 | 0.184 | 0.203 | 0.116 | 0.002 | n/c | |
8带纹条鳎 Zebrias zebra | 0.245 | 0.245 | 0.244 | 0.277 | 0.278 | 0.302 | 0.304 | 0.002 |
图1 基于K2P进化距离构建的各种类NJ进化树。节点处的数字为1,000次bootstrap自展检验支持率(只显示大于50%的数值)。图中序列编号同表2。
Fig. 1 Neighbor-joining tree constructed using the K2P model for COI gene sequences of eight species. The numbers indicate the percentages of bootstrap values (>50%) supporting each node from 1,000 replicates. The Sequence numbers are the same to Table 2.
图2 舌鳎属样品(Cynoglossus sp.)鱼卵及仔鱼的形态结构。(a)胚胎绕卵黄囊1/2; (b)初孵仔鱼, 体长1.16 mm; (c)后期仔鱼, 体长6.21 mm。
Fig. 2 Morphological characters of egg and larva of Cynoglossus sp. (a) Embryo ring 1/2 epiboly; (b) 1.16 mm BL newly hatched larva; (c) 6.21 mm BL post-metamorphosis larva.
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