phylogenetic position of the altaic fish genus oreoleuciscus in leuciscinae based on mitogenome...
TRANSCRIPT
Phylogenetic position of the Altaic fish
genus Oreoleuciscus in Leuciscinae
based on mitogenome sequences
Junichi ImotoJunichi Imoto11, Kenji Saitoh, Kenji Saitoh22, Jun Adachi, Jun Adachi33, Takeshi Sasaki, Takeshi Sasaki44, , Yuri P. KartavtsevYuri P. Kartavtsev55, Satoru N. Chiba, Satoru N. Chiba11, Ryo O. Gotoh, Ryo O. Gotoh11, , Masaki MiyaMasaki Miya55, Mutsumi Nishida, Mutsumi Nishida66 and Naoto Hanzawa and Naoto Hanzawa11
11Grad Sch Sci Eng, Yamagata Univ, Grad Sch Sci Eng, Yamagata Univ, 22Nat Res Inst Fish SciNat Res Inst Fish Sci, , 33Inst Stat Math, Inst Stat Math, 44Inst Marine Biol, Inst Marine Biol, 55Nat Hist Mus Inst Chiba, Nat Hist Mus Inst Chiba,
66Ocean Res Inst, Univ TokyoOcean Res Inst, Univ Tokyo
Phylogenetic position of the Altaic fish
genus Oreoleuciscus in Leuciscinae
based on mitogenome sequences
38 species 204 genera (FishBase)leuciscin: Eurasian plus North American Notemigonus cryleucas
48 species 319 genera (FishBase)phoxinin: North American and Far Eastern species
Phylogeny based on morphology (Cavender and Coburn, 1992)
Leuciscinae
Widely distributed in Northern Hemisphere
Morphologically similar (even among different genera)
Important to consider the evolutional history of Leuciscinae
Central Asia and Far East
The genus Oreoleuciscus (Warpachowskii, 1889)
Endemic to inland lakes in western Mongolia and Tuva, the upper Ob River system within the Altai Mountain region of Siberia (Berg, 1962)
Planktonic algae, insects, invertebrates and fishes
Oreoleuciscus potanini
Feeding habitat
Distribution
Far E
asternE
uropeanN
orth Am
erican
ML tree based on mt genome
Phylogenetic position of Oreoleuciscus
Oreoleucuscus potanini
Closely related to Pseudaspius and Tribolodon ? (Howes, 1984, 1991)Tribolodon
sachalinensis
Tribolodon hakonensis
Pseudaspius leptocephalus
?
Tribolodon nakamurai
Far E
asternE
uropeanN
orth Am
erican
ML tree based on mt genome
Phylogenetic position of Oreoleuciscus
Oreoleucuscus potanini
Closely related to Phoxinus ? (Travers, 1989; Bogutskaya, 1990)
Phoxinus phoxinusPhoxinus lagowskii Phoxinus perenurus
?
No molecular phylogenetic analyses for Oreoleuciscus species
High degree of morphological variability that obscures differences among species
Taxonomy of the genus Oreoleuciscus is controversial
Paucity of specimens from key geographical areas
Determine complete mitochondrial (mt) genome sequences of Oreoleuciscus
Conduct phylogenetic analysis based on mt genome sequences
Clarify the phylogenetic position of genus Oreoleuciscus and evolutional history of the Leuciscinae
The purpose of this study
Mt genome analyses
Long DNA sequences
(Inoue et al., 2001a; Miya et al., 2003; Saitoh et al., 2006)
Correct estimation of phylogeny at various taxonomic levels
with strong statistical support
17 genera 31 species
Outgroup 9 species (Cypridae 8, Cobitidae 1)
ML RAxML v7.0.4 (Stamatakis, 2006)
Bayes MrBayes v3.1.2 (Ronquist & Huelsenbeck 2003)
Determined mt genome sequences
Genomic structure Common with vertebrates
Exclude ND6, control region and gaps (14,826 bp)
Mt genome of Oreoleuciscus potanini
0.1
Phoxinus perenurus mantschricus Vladivostok RussiaPhoxinus perenurus sachalinensis Hokkaido Japan
100
Phoxinus perenurus Poland
100
Phoxinus oxycephalus jouyi Kochi JapanPhoxinus lagowskii Mongol
Phoxinus lagowskii steindachneri Mogami River Japan100
100
Oreoleuciscus potanini Altai
100
Tribolodon brandtii Ukedo River JapanTribolodon brandtii Mogami River Japan
Tribolodon nakamurai Mogami RIver Japan
100
Pseudaspius leptocephalus Lake Bolony RussiaTribolodon sachalinensis Tadami River JapanTribolodon hakonensis Mogami River Japan
100
100
Phoxinus phoxinus MongolPhoxinus phoxinus Italy100
Cyprinella lutrensisCyprinella spiloptera
100
Notropis stramineus
100
Phenacobius mirabilis
100
Campostoma anomalum
100
Mylocheilus caurinus
100
Gila robustaGila conspersa
100
Phoxinus eos
100
Alburnus alburnusLeucaspius delineatus
100
Notemigonus crysoleucasAbramis brama
Blicca bjoerkna100
Chondrostoma lemmingii
100
Pelecus cultratus
100
100
Tinca tincaPseudorasbora pumila
Sarcocheilichthys variegatus microoculus
100
Megalobrama terminalisXenocypris argentea
10
Far E
asternE
uropeanN
orth Am
erican
ML bootstrap / Bayesian posterior probabilities
0.1
Phoxinus perenurus mantschricus Vladivostok RussiaPhoxinus perenurus sachalinensis Hokkaido Japan
100
Phoxinus perenurus Poland
100
Phoxinus oxycephalus jouyi Kochi JapanPhoxinus lagowskii Mongol
Phoxinus lagowskii steindachneri Mogami River Japan100
100
Oreoleuciscus potanini Altai
100
Tribolodon brandtii Ukedo River JapanTribolodon brandtii Mogami River Japan
Tribolodon nakamurai Mogami RIver Japan
100
Pseudaspius leptocephalus Lake Bolony RussiaTribolodon sachalinensis Tadami River JapanTribolodon hakonensis Mogami River Japan
100
100
Phoxinus phoxinus MongolPhoxinus phoxinus Italy100
Cyprinella lutrensisCyprinella spiloptera
100
Notropis stramineus
100
Phenacobius mirabilis
100
Campostoma anomalum
100
Mylocheilus caurinus
100
Gila robustaGila conspersa
100
Phoxinus eos
100
Alburnus alburnusLeucaspius delineatus
100
Notemigonus crysoleucasAbramis brama
Blicca bjoerkna100
Chondrostoma lemmingii
100
Pelecus cultratus
100
100
Tinca tincaPseudorasbora pumila
Sarcocheilichthys variegatus microoculus
100
Megalobrama terminalisXenocypris argentea
10
Far E
asternE
uropeanN
orth Am
erican
ML bootstrap / Bayesian posterior probabilities
0.1
Phoxinus perenurus mantschricus Vladivostok RussiaPhoxinus perenurus sachalinensis Hokkaido Japan
100
Phoxinus perenurus Poland
100
Phoxinus oxycephalus jouyi Kochi JapanPhoxinus lagowskii Mongol
Phoxinus lagowskii steindachneri Mogami River Japan100
100
Oreoleuciscus potanini Altai
100
Tribolodon brandtii Ukedo River JapanTribolodon brandtii Mogami River Japan
Tribolodon nakamurai Mogami RIver Japan
100
Pseudaspius leptocephalus Lake Bolony RussiaTribolodon sachalinensis Tadami River JapanTribolodon hakonensis Mogami River Japan
100
100
Phoxinus phoxinus MongolPhoxinus phoxinus Italy100
Cyprinella lutrensisCyprinella spiloptera
100
Notropis stramineus
100
Phenacobius mirabilis
100
Campostoma anomalum
100
Mylocheilus caurinus
100
Gila robustaGila conspersa
100
Phoxinus eos
100
Alburnus alburnusLeucaspius delineatus
100
Notemigonus crysoleucasAbramis brama
Blicca bjoerkna100
Chondrostoma lemmingii
100
Pelecus cultratus
100
100
Tinca tincaPseudorasbora pumila
Sarcocheilichthys variegatus microoculus
100
Megalobrama terminalisXenocypris argentea
10
Far E
asternE
uropeanN
orth Am
erican
ML bootstrap / Bayesian posterior probabilities
Oreoleuciscus potanini
Phoxinus lagowskii
Tribolodon nakamurai
0.1
Phoxinus perenurus mantschricusPhoxinus perenurus sachalinensis
99
Phoxinus perenurus
100
Phoxinus lagowskii steindachneriPhoxinus lagowskii
10031
Phoxinus oxycephalus jouyi
100
Oreoleuciscus potanini
100
Tribolodon brandtii Ukedo RiverTribolodon brandtii Mogami River
68
Tribolodon nakamurai
100
Pseudaspius leptocephalus
74
Tribolodon sachalinensis
97
Tribolodon hakonensis100
100
Phoxinus phoxinus ItalyPhoxinus phoxinus Mongol100
97
Cyprinella lutrensisCyprinella spiloptera100
Notropis stramineus
100
Phenacobius mirabilis
100
Campostoma anomalum
100
Mylocheilus caurinus
100
86
Gila robustaGila conspersa
100
Phoxinus eos
100
80
Alburnus alburnusLeucaspius delineatus
100
Notemigonus crysoleucas
66
Blicca bjoerknaAbramis brama
100
Chondrostoma lemmingii21
100
Pelecus cultratus100
100
Tinca tinca
95
Sarcocheilichthys variegatus mPseudorasbora pumila
100
85
Megalobrama terminalisCulter mongolicus
100
Xenocypris argentea
100
Zacco sieboldtii100
100
Labeo batesiiCyprinus carpio
100
100
Misgurnus nikolskyiCobitis striata
100
Lefua echigonia
100
100
Engraulis japonicusSardinops melanostictus
100
100
Astronotus ocellatusTropheus duboisi
46
Paratilapia polleni
100
Etroplus maculatusParetroplus maculatus
100100
Oryzias latipes
100
Takifugu rubripesTetraodon nigroviridis
100
43
Paralichthys oliva
L 100, U 120
L 112, U 145
L 85, U 95
U 363
Molecular Dating
(Mya)
Upper constraint : U
Lower constraint : L
Parametric Bayesian approach
Multidistribute (Thorn & Kishino 2002)
Cichlids
L 100, U 120
L 112, U 145
L 85, U 95
African
South American
Madagascar
Indo/Sri Lanka
Madagascar
Cichlid
Molecular Dating
(Mya)
Upper constraint : U
Lower constraint : L
(Azuma et al., 2008)
Phoxinus perenurus mantschricusVladivostok RussiaPhoxinus perenurus sachalinensisHokkaidao JapanPhoxinus perenurus PolandPhoxinus lagowskii MongolPhoxinus lagowskii steindachneriMogami River JapanPhoxinus oxycephalus jouyi Kochi JapanOreoleuciscus potanini AltaiTribolodon brandtii Ukedo River JapanTribolodon brandtii Mogami River JapanTribolodon nakamurai Mogami RIver JapanPseudaspius leptocephalus Lake Bolony RussiaTribolodon sachalinensis Tadami River JapanTribolodon hakonensis Mogami River JapanPhoxinus phoxinus MongolPhoxinus phoxinus ItalyCyprinella lutrensisCyprinella spilopteraNotropis stramineusPhenacobius mirabilisCampostoma anomalumMylocheilus caurinusGila robustaGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaBlicca bjoerknaChondrostoma lemmingiiPelecus cultratusTinca tincaPseudorasbora pumilaSarcocheilichthys variegatus microoculusCulter mongolicusMegalobrama terminalisXenocypris argenteaZacco sieboldtiiCyprinus carpioLabeo batesii
Leuciscinae / other subfamily
Cyprinidae / Cobitidae
331.2
Otocephala / Euteleostei
219.2
123.6
Mesozoic CenozoicPaleozoic
Carboniferous
Triassic Jurassic Cretaceous TertiaryPermian
Phoxinus perenurus mantschricusVladivostok RussiaPhoxinus perenurus sachalinensisHokkaidao JapanPhoxinus perenurus PolandPhoxinus lagowskii MongolPhoxinus lagowskii steindachneri Mogami River JapanPhoxinus oxycephalus jouyi Kochi JapanOreoleuciscus potanini AltaiTribolodon brandtii Ukedo River JapanTribolodon brandtii Mogami River JapanTribolodon nakamurai Mogami RIver JapanPseudaspius leptocephalus Lake Bolony RussiaTribolodon sachalinensis Tadami River JapanTribolodon hakonensis Mogami River JapanPhoxinus phoxinus MongolPhoxinus phoxinus ItalyCyprinella lutrensisCyprinella spilopteraNotropis stramineusPhenacobius mirabilisCampostoma anomalumMylocheilus caurinusGila robustaGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaBlicca bjoerkna
phoxinin / leuciscin
84.8
Far Eastern phoxinin / P. phoxinus
78.5
Orieoleuciscus / Far Eastern Phoxinus35.5
Tribolodon / Orieoleuciscus + Far Eastern Phoxinus48.6
Mesozoic Cenozoic
Cretaceous Pa. Eocene Oli. Miocene Plio
The genus Oreoleuciscus is included in phoxinin (Cavender & Coburn 1992)
rather than Tribodolon and Pseudaspius
Oreoleuciscus formed a monophyletic clade with the Far Eastern phoxinins
Conclusion
Oreoleuciscus is closely related to the Far Eastern Phoxinus
Tribolodon, Pseudaspius and the Far Eastern Phoxinus
Tribolodon + Pseudaspius and Oreoleuciscus + Phoxinus were divided 49 Mya
Oreoleuciscus and Phoxinus were divided 34 Mya
Thank you for your attention !!
The genus Oreoleuciscus (Warpachowskii, 1889)
Endemic to inland lakes in western Mongolia and Tuva, the upper Ob River system within the Altai Mountain region of Siberia (Berg, 1962)
The range of Oreoleuciscus demarcates a highly ranked biogeographical unit that has a very depauperate and distinct ichthyofauna.
In the review of Eurasian cyprinid biogeography, Banarescu & Coad (1991) determined that the rank of this unit is equal to that of regions such as the entire Euro-Mediterranean or Central Asian regions.
Oreoleucuscus potanini
Far E
asternE
uropeanN
orth Am
erican
1.81 ~ 4.37 Mya Tribolodon speciation
The glacial period
12.90 ~ 15.41 Mya Radiation of Leuciscinae
Rapid global cooling of Middle Miocene
The sea level dropped
The land area expansion, freshwater and brackish water area increased
Environment habitable for Leuciscinae extended
Enlarge their distribution and occurred speciation
Far E
asternE
uropeanN
orth Am
erican
ML tree based on mt genome
Ancestral area of the Far Eastern Leuciscinae
The genus Oreoleuciscus (Warpachowskii, 1889)
Endemic to inland lakes in western Mongolia and Tuva, the upper Ob River system within the Altai Mountain region of Siberia (Berg, 1962)
The range of Oreoleuciscus demarcates a highly ranked biogeographical unit that has a very depauperate and distinct ichthyofauna.
In the review of Eurasian cyprinid biogeography, Banarescu & Coad (1991) determined that the rank of this unit is equal to that of regions such as the entire Euro-Mediterranean or Central Asian regions.
10
Tribolodon brandtii UKETribolodon brandtii MGMTribolodon nakamuraiPseudaspius leptocephalusTribolodon sachalinensisTribolodon hakonensisPhoxonus perenurus sachalinensisPhoxonus perenurus mantschricusPhoxonus perenurusPhoxinus oxycephalus jouyiPhoxinus lagowskii steindachneriPhoxinus lagowskiiPhoxinus phoxinus MONPhoxinus phoxinus ITACyprinella lutrensisNotropis stramineusMylocheilus caurinusGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaPelecus cultratusTinca tincaSarcocheilichthys variegatus microoculusMegalobrama treminalisXenocypris argenteaCyprinus carpioLabeo batesiiCobitis striataLefua echigoniaEngraulis japonicusSardinops melanostictusAstronotus ocellatusTropheus duboisiParatilapia polleniEtroplus maculatusParetroplus maculatusOryzias latipesParalichthys olivaceusTakifugu rubripesTetraodon nigroviridisGasterosteus aculeatus
100 0200
中世代
第三紀
第四紀古世代
白亜紀ジュラ紀三畳紀ペルム紀
(Mya)
ウグイ亜科と他の亜科
コイ科とドジョウ科
271.0
ニシン・骨鰾下区と正新骨下区
189.7
100.3
化石記録よりも古い年代
コイ科魚類 最古の化石Parabarbus sp. ca. 50
Mya
24
ユーラシアおよび北米に広く分布するコイ科ウグイ亜科魚類は、主にユーラシアに分布する leuciscin と、主に北米および極
東に分布する phoxinin の 2 つの系統に大別される。
ウグイ亜科魚類の系統地理学的考察を行うためには、 2 つの系統群が接する極東から中央ユーラシアにかけて分布するタクソ
ンのサンプリングが必要と考えられ、発表者らは、アルタイ共和国においてサンプリングを行い、ウグイ亜科 Oreoleuciscus
属魚類を採集した ( 千葉ら , 2008) 。
Oreoleuciscus 属魚類は中央アジア・オビ川上流のアルタイ山地、モンゴルの一部にのみ分布する非常に希少な魚類である。
Howes (1984, 1991) は、形態的特徴から Oreoleuciscus 属は Pseudaspius 属やウグイ属と共に aspinin の系統に含まれる
とした。
Howes (1984, 1991) ウグイ亜科を Abramin 、 Aspinin 、 Phoninin の 3 つに分類
しかし、 Oreoleuciscus 属は形態的に Phoxinus 属に近いという考えもあり (Travers, 1989; Bogutskaya, 1990) 、未だ
に系統的位置は不明だが、これまで分子系統解析は全く行われていない。
そこで、我々は Oreoleuciscus potanini のミトコンドリアゲノムを決定して、他のウグイ亜科魚類とともに分子系統解析を
行った。
その結果、 Oreoleuciscus 属は極東産 Phoxinus 属の姉妹群となり、これらにウグイ属を加えたクレードは単系統群を形成し、
他の欧米産ウグイ亜科魚類各クレードからは大きく離れていた。
本研究の結果は、ウグイ亜科魚類の進化史を考察する上で重要なデータと考えられる。
10
Tribolodon brandtii UKETribolodon brandtii MGMTribolodon nakamuraiPseudaspius leptocephalusTribolodon sachalinensisTribolodon hakonensisPhoxonus perenurus sachalinensisPhoxonus perenurus mantschricusPhoxonus perenurusPhoxinus oxycephalus jouyiPhoxinus lagowskii steindachneriPhoxinus lagowskiiPhoxinus phoxinus MONPhoxinus phoxinus ITACyprinella lutrensisNotropis stramineusMylocheilus caurinusGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaPelecus cultratusTinca tincaSarcocheilichthys variegatus microoculusMegalobrama treminalisXenocypris argenteaCyprinus carpioLabeo batesiiCobitis striataLefua echigoniaEngraulis japonicusSardinops melanostictusAstronotus ocellatusTropheus duboisiParatilapia polleniEtroplus maculatusParetroplus maculatusOryzias latipesParalichthys olivaceusTakifugu rubripesTetraodon nigroviridisGasterosteus aculeatus
100 0200
中世代
第三紀
第四紀古世代
白亜紀ジュラ紀三畳紀ペルム紀
(Mya)
コイ科とドジョウ科
271.0
ニシン・骨鰾下区と正新骨下区
189.7
化石記録よりも古い年代
コイ科魚類 最古の化石Parabarbus sp. ca. 50
Mya分子を用いた年代推定とほぼ一致
(Inoue et al., 2005; Yamanoue et al., 2006; Azuma et al., 2008)
10
Tribolodon brandtii UKETribolodon brandtii MGMTribolodon nakamuraiPseudaspius leptocephalusTribolodon sachalinensisTribolodon hakonensisPhoxonus perenurus sachalinensisPhoxonus perenurus mantschricusPhoxonus perenurusPhoxinus oxycephalus jouyiPhoxinus lagowskii steindachneriPhoxinus lagowskiiPhoxinus phoxinus MONPhoxinus phoxinus ITACyprinella lutrensisNotropis stramineusMylocheilus caurinusGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaPelecus cultratusTinca tincaSarcocheilichthys variegatus microoculusMegalobrama treminalisXenocypris argenteaCyprinus carpioLabeo batesiiCobitis striataLefua echigoniaEngraulis japonicusSardinops melanostictusAstronotus ocellatusTropheus duboisiParatilapia polleniEtroplus maculatusParetroplus maculatusOryzias latipesParalichthys olivaceusTakifugu rubripesTetraodon nigroviridisGasterosteus aculeatus
中世代 新生代
白亜紀ジュラ紀
暁新生
始新生
漸新世
中新生
鮮新世
100 0 (Mya)
phoxinin / leuciscin
74.7
Far Eastern phoxinin / P. phoxinus
61.7
Orieoleuciscus / Far Eastern Phoxinus
21.2Tribolodon / Orieoleuciscus + Far Eastern Phoxinus
39.2
イタリアモンゴル
最上川請戸川
10
Tribolodon brandtii UKETribolodon brandtii MGMTribolodon nakamuraiPseudaspius leptocephalusTribolodon sachalinensisTribolodon hakonensisPhoxonus perenurus sachalinensisPhoxonus perenurus mantschricusPhoxonus perenurusPhoxinus oxycephalus jouyiPhoxinus lagowskii steindachneriPhoxinus lagowskiiPhoxinus phoxinus MONPhoxinus phoxinus ITACyprinella lutrensisNotropis stramineusMylocheilus caurinusGila conspersaPhoxinus eosAlburnus alburnusLeucaspius delineatusNotemigonus crysoleucasAbramis bramaPelecus cultratusTinca tincaSarcocheilichthys variegatus microoculusMegalobrama treminalisXenocypris argenteaCyprinus carpioLabeo batesiiCobitis striataLefua echigoniaEngraulis japonicusSardinops melanostictusAstronotus ocellatusTropheus duboisiParatilapia polleniEtroplus maculatusParetroplus maculatusOryzias latipesParalichthys olivaceusTakifugu rubripesTetraodon nigroviridisGasterosteus aculeatus
中世代 新生代
白亜紀ジュラ紀
暁新生
始新生
漸新世
中新生
鮮新世
100 0 (Mya)
7.5 ~ 21.2 Mya Tribolodon 属各種が種分化
寒冷化・乾燥化
陸地が拡大、淡水域や汽水域などが増える
ウグイ亜科魚類が生息できる環境が広がる
新しい生息場所に分布を広げながら種分化
海水面の低下
7.5
21.2
イタリアモンゴル
最上川請戸川
Closely related to Pseudaspius and Tribolodon ? (Howes, 1984, 1991)
Closely related to Phoxinus ? (Travers, 1989; Bogutskaya, 1990)
Tribolodon sachalinensis
Tribolodon hakonensis
Tribolodon brandtii
Tribolodon nakamurai
Phoxinus phoxinusPhoxinus lagowskii Phoxinus perenurus
Pseudaspius leptocephalus
High degree of morphological variability that obscures differences among species
Taxonomy of the genus Oreoleuciscus is controversial
Paucity of specimens from key geographical areas
Phylogenetic position of Oreoleuciscus
Oreoleuciscus is thought to be allied to the Eurasian cyprinid genera Pseudaspius and Tribolodon which are within the aspinin lineage of the subfamily Leuciscinae (Howes, 1984, 1991).
On the other hand, there is also the idea that Oreoleuciscus is rather closely related to the phoxinin lineage (Travers, 1989; Bogutskaya, 1990).
Thus, taxonomy of the genus Oreoleuciscus is controversial, in part because of the paucity of specimens from key geographical areas, and in part because there is high degree of morphological variability that obscures differences among species.
Determine complete mitochondrial (mt) genome sequences of Leuciscinae species
Purpose
Conduct phylogenetic analysis based on mt genome sequences
Clarify the phylogenetic position of genus Oreoleuciscus and evolutional history of the Leuciscinae
Determine complete mitochondrial (mt) genome
sequences of Leuciscinae species
Purpose
Conduct phylogenetic analysis based on
mt genome sequences
Clarify the phylogenetic position of genus Oreoleuciscus
and evolutional history of the Leuciscinae
38 species 204 genera (FishBase)leuciscin: Eurasian plus North American Notemigonus cryleucas
48 species 319 genera (FishBase)phoxinin: North American and Far Eastern species
Phylogeny based on morphology (Cavender and Coburn, 1992)
Leuciscinae
33
ウグイ亜科の分類 phoxinin と leuciscin
中央アジアが Point だけどサンプルが少ない
ウグイ亜科の進化史を考えると重要
今回アルタイで Oreoleuciscus を採集
Oreoleuciscus はどこに入るか ?
T. sachalinensis
T. hakonensis
T. brandtii
T. nakamurai
Low level of morphological differentiation
Their taxonomy had been contentious (1 ~ 5 species).
Tribolodon
(Ikeda, 1937; Nakamura, 1963,1969; Onodera & Honma, 1976; Kurawaka, 1977)
Endemic in the Far East
Far Eastern phoxinin
Inhabit the upper reaches of rivers, lakes and coastal regions of the sea
The pharyngeal teeth of Pseudaspius and Tribolodon species are similar.
Pseudaspius leptocephalus
P. leptocephalus T. nakamurai T. sachalinensis
(Tomoda, 1998)
Previous molecular phylogenetic studies(Kartavtsev et al., 2002; Sakai et al., 2002, 2006; Sasaki et al., 2007)
Pseudaspius and Tribolodon species are closely related
in the Far Eastern phoxinin branch.
Widely distributed in Northern Hemisphere (Nelson, 2006)
Body size small
Classified into a single genus “Phoxinus”
7 species live in the Far East
Degree of morphological differentiation is lower than the other genera
P. phoxinusP. lagowskii P. perenurus
Phoxinus
Previous molecular phylogenetic studies (Sasaki et al., 2007)
Far Eastern Phoxinus species and P. phoxinus are polyphyletic
Far Eastern phoxinin
Phylogenetic analysis based on cyt b gene
cyt b gene sequences 1140 bp
Useful to estimate phylogenetic relationships in closely related species
Large number of sequence are registered in databases.
North American27 species
Far Eastern30 species
European36 species
Outgroup 9 species (Cypridae 8, Cobitidae 1)
Neighbor Joining (NJ) Analysis
45 genera 93 species
NJ tree based on cyt b gene
Far Eastern phoxinin
North American phoxinin
North American phoxinin
European leuciscin
&North American Notemigonus
North American phoxinin
Phoxinus phoxinus
Relationship among clades
Unclear
NJ tree based on cyt b gene
Phylogenetic analysis based on mt genome
mt genome analysis
Long DNA sequences
(Inoue et al., 2001a; Miya et al., 2003; Saitoh et al., 2006)
Determined mt genome sequences ca. 16,600 bp (circular)
Genomic structure common with vertebrates
Exclude ND6, control region and gaps (14,969 bp)
mt genome of Leuciscinae species
North American10 species
Far Eastern13 species
European7 species
Selected species from major clades in the cyt b-based tree
16 genera 30 species
Outgroup 9 species (Cypridae 8, Cobitidae 1)
Maximum likelihood (ML) Analysis (RAxML)
Correct estimation of phylogeny at various taxonomic levels
with strong statistical support
Far E
asternE
uropeanN
orth Am
erican
ML tree based on mt genome
ML tree based on mt genome gene
Far E
asternE
uropeanN
orth Am
erican
ML tree based on mt genome
phoxinin
leuciscin
Far E
asternE
uropeanN
orth Am
erican
This analysis supports the classification by Cavender and Coburn (1992)
Divided into 2 clades; phoxinin & leuciscin
Far E
asternE
uropeanN
orth Am
erican
The Far Eastern phoxinins Tribolodon, Pseudaspius, Phoxinus
Monophyletic and greatly diverged from the other Leuciscinae clades
Far E
asternE
uropeanN
orth Am
erican
Pseudaspius leptocephalus
Tribolodon brandtii
Tribolodon species and Pseudaspius leptocephalus
A single genus ?
Subclade with 100% statistical supports
Far E
asternE
uropeanN
orth Am
erican
North American species
Divided into some independent clades
Notemigonus cryleucas European leuciscin clade
Far E
asternE
uropeanN
orth Am
erican
Far Eastern Phoxinus species, European P. phoxinus, North American P. eos
P. eos
P. lagowskii
P. phoxinus
Polyphyletic Some different genera ?
Far E
asternE
uropeanN
orth Am
erican
mt genome tree
Divergence time estimation
L 0.014 Mya
L 0.014 Mya
North America isolation from Eurasia
The oldest and reliable fossils of Cyprinidae (Cavender, 1991)
Fossil record of Pseudorasbora (Liu and Su, 1962)
U 54.8 L 49.0 Mya
L 3.0 Mya
Lower constraint : L
Upper constraint : U
Bayesian method; mcmctree (PAML 4; Yang and Rannala, 2006)
15.14 Mya
12.90 Mya
8.82 Mya
4.37 Mya
19.74 Mya
Far E
asternE
uropeanN
orth Am
erican
Far E
asternE
uropeanN
orth Am
erican
1.81 ~ 4.37 Mya Tribolodon speciation
The glacial period
12.90 ~ 15.41 Mya Radiation of Leuciscinae
Rapid global cooling of Middle Miocene
The sea level dropped
The land area expansion, freshwater and brackish water area increased
Environment habitable for Leuciscinae extended
Enlarge their distribution and occurred speciation
(Miya and Nishida, 1999)
Long PCR
Cycle sequencing
Same primers as for the nested PCR
Nested PCR
162 Fish-versatile primers55 cyprinid-specific primers
Determination of whole mt genome sequences
4 combinations of Fish-versatile primers