全 文 :植物科学学报 2014ꎬ 32(3): 216 ~ 227
Plant Science Journal
DOI:10 3724 / SP J 1142 2014 30216
基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究
钟彩霞1ꎬ 黎维平1∗ꎬ 杨秀林1ꎬ 唐 明2ꎬ 廖 威1ꎬ 陈三茂1
(1 湖南师范大学生命科学学院ꎬ 长沙 410081ꎻ 2 中国科学院华南植物园ꎬ
中国科学院植物资源保护与可持续利用重点实验室ꎬ 广州 510650)
摘 要: 国产白酒草亚族(菊科紫菀族)由白酒草属(Conyza)、 小舌菊属(Microglossa)和歧伞菊属(Thespis)
3个小属组成ꎬ 且国产白酒草亚族各属间及其与非洲白酒草属植物之间的分子系统发育关系尚无报道ꎬ 故本研究
利用核糖体 DNA ITS和 ETS序列并采用最大简约法和贝叶斯分析法ꎬ 重建了国产白酒草亚族的分子系统发育
树ꎮ 结果表明ꎬ 国产 4种白酒草属植物、 歧伞菊和非洲白酒草属植物组成一支ꎬ 而劲直白酒草的两变种和小舌
菊嵌入田基黄亚族分支ꎻ 小舌菊与 Psiadia pascalii近缘ꎮ 基于这些结果ꎬ 我们认为: (1)劲直白酒草和 Conyza
incisa应处理为田基黄亚族的一个独立的属ꎻ (2)国产 4种白酒草属植物和歧伞菊以及大多数非洲白酒草属植物
属于 Eschenbachia属ꎬ 而且 Eschenbachia属代表一个新的亚族ꎬ 歧伞菊可处理为 Eschenbachia属的一个组ꎮ
Eschenbachia属可能从非洲经数次长距离传播到达我国南部ꎻ (3)Welwitschiella 和小舌菊属应保持属的地位ꎬ
Psiadia pascalii、 Conyza scabrida和 C pyrrhopappa可并入小舌菊属ꎮ
关键词: 国产白酒草族ꎻ 白酒草属ꎻ Eschenbachiaꎻ 小舌菊属ꎻ 分子系统发育ꎻ 歧伞菊属
中图分类号: Q949 783 5 文献标识码: A 文章编号: 2095 ̄0837(2014)03 ̄0216 ̄12
收稿日期: 2014 ̄01 ̄21ꎬ 修回日期: 2014 ̄02 ̄20ꎮ
基金项目: 国家自然科学基金资助项目(31370265)ꎻ 湖南省生态学重点学科建设项目资助ꎻ 湖南省教育厅科学研究基金资助
(08A046)ꎮ
钟彩霞(1988-)ꎬ 女ꎬ 硕士研究生ꎬ 主要研究方向为种子植物分类学(E ̄mail: 860905430@ qq com)ꎮ
∗ 通讯作者(Author for correspondence. E ̄mail: lwprenyi@ aliyun com)ꎮ
Molecular Phylogeny of Chinese Conyzaꎬ Microglossa and
Thespis (Asteraceae: Astereae) Based on
Two Nuclear Ribosomal DNA Regions
ZHONG Cai ̄Xia1ꎬ LI Wei ̄Ping1∗ꎬ YANG Xiu ̄Lin1ꎬ TANG Ming2ꎬ LIAO Wei1ꎬ CHEN San ̄Mao1
(1 College of Life Sciencesꎬ Hunan Normal Universityꎬ Changsha 410081ꎬ Chinaꎻ
2 Key Laboratory of Plant Resources Conservation and Sustainable Utilizationꎬ South China Botanical Gardenꎬ
Chinese Academy of Sciencesꎬ Guangzhou 510650ꎬ China)
Abstract: Chinese subtribe Conyzinae (Asteraceae: Astereae) consists of three small
generaꎬ Conyzaꎬ Microglossa and Thespis To dateꎬ howeverꎬ the molecular phylogenetic
relationships among African Conyzaꎬ and Chinese Conyzaꎬ Thespis and Microglossa have
not been investigated The internal and external transcribed spacers of nuclear ribosomal DNA
were used to reconstruct the phylogeny of Chinese Conyzinae through maximum parsimony
and Bayesian analyses Results showed that four Chinese Conyza species and Thespis
divaricata belonged to a clade with most sampled African Conyzaꎬ whereas two Conyza
stricta varieties and Microglossa pyrifolia were embedded deeply within the subtribe
Grangeinae clade Microglossa pyrifolia was closely related to Psiadia pascalii Based on
phylogenyꎬ we considered that (1) C stricta and C incisa should be treated as a separate
genus that belongs to subtribe Grangeianeꎻ (2) four Chinese Conyzaꎬ Thespis and most
sampled African Conyza belong to the genus Eschenbachia Eschenbachia should belong to a
new separate subtribeꎬ while Thespis should be treated as a section of Eschenbachia
Eschenbachia may have arrived at southern China by several long ̄distance dispersals from
Africaꎻ (3) it is reasonable to maintain the generic status of Welwitschiella and Microglossa
and treat Psiadia pascaliiꎬ C scabrida and C pyrrhopappa as members of Microglossa.
Key words: Chinese Conyzinaeꎻ Conyzaꎻ Eschenbachiaꎻ Microglossaꎻ Molecular phylogenyꎻ
Thespis
Although tribe Astereae is the second largest
in Asteraceae (Compositae)ꎬ phylogenetic rela ̄
tionships within Astereae are still far from re ̄
solved[1 ̄5] Chinese Astereae subtribe Conyzinae
consists of three generaꎬ Conyza Lessꎬ Micro ̄
glossa DC and Thespis DC [6]ꎬ which have not
yet been phylogenetically evaluated based on
molecular data.
Conyza was established in 1832 by Lessing
and typified by C chilensis Sprengelꎬ a New
World species ( = Conyza primulifolia (Lam)
Cuatrec and Lourteig) [6ꎬ7] Morphological studi ̄
es[7 ̄10] and molecular data[1ꎬ2ꎬ4ꎬ11 ̄13] revealed that
New World Conyza was not closely related to Old
World Conyza Furthermoreꎬ phylogenetic analy ̄
ses[1ꎬ2ꎬ12] suggested that New World Conyza was
at least biphyletic and northern and southern
American Conyza were all derived from Erigeron
L Accordinglyꎬ Chen et al[14] merged a few
New World Conyza species introduced to China
into Erigeron Converselyꎬ Nesom[7] suggested
that Old World species identified as Conyza were
related to genera outside Conyzinaeꎬ and Ne ̄
som and Robinson[10] stated that “ species of
Conyza native to Africa apparently are more simi ̄
lar and perhaps more closely related to genera of
subtribe Grangeinae” Similarlyꎬ Brouillet et al [1]
suggested that the subtribe Grangeinae should
be expanded to include African Conyza Re ̄
centlyꎬ as a small part of Old World Conyzaꎬ
Conyza species ( excluding C stricta Willd )
native to China were assigned to Eschenbachia
Moench[14]ꎬ the oldest name representing Old
World Conyza[8] .
Howeverꎬ Old World Conyza is not mono ̄
phyletic As early as 1990ꎬ based on certain
morphological featuresꎬ Nesom[8] concluded that
Old World Conyza was strongly heterogeneous
and its taxonomy was far from resolved In addi ̄
tionꎬ some molecular phylogenetic analyses
have supported Nesom[8]ꎬ showing that Old
World Conyza was polyphyletic[3] or at least
biphyletic[1ꎬ2ꎬ11ꎬ13]. For exampleꎬ 14 sampled
Conyza species of Madagascar and Indian
Ocean islands formed several smallerꎬ mostly
weakly supported clades with two to five species
in phylogenetic treesꎬ indicating that African
Conyza was polyphyletic[3] Strijk[3] transferred
three African Conyza species (C scabrida DCꎬ
C pyrrhopappa Sch Bip and C ageratoides
DC ) to a newly circumscribed Psiadia s sꎬ
whereas other African Conyza species were
paraphyletic with respect to Frappieria Cordemoyꎬ
which was resurrected by Strijk[3] to accommo ̄
date the smaller of the two clades of Psiadia.
Up to nowꎬ howeverꎬ molecular phyloge ̄
netic analysesꎬ including Old World Conyzaꎬ
have mainly sampled African species[1 ̄5ꎬ11ꎬ13] .
Three Conyza speciesꎬ C blinii H Lév (Es ̄
chenbachia blinii (H Lév ) Brouillet )ꎬ C
perennis Hand  ̄Mazz (Eschenbachia perennis
(Hand  ̄Mazz ) Brouillet) and C muliensis Y
L Chen (Eschenbachia muliensis (Y L Chen)
Brouillet)ꎬ are endemic to Chinaꎬ though their
molecular data have not yet been reported.
Thespis DC is a small genus with three spe ̄
712
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钟彩霞等: 基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究(英文)
cies and distributed in south ̄eastern Asia Thes ̄
pis divaricata DC is the only species of the ge ̄
nus from China The systematic position of this
genus has been problematic for a long time Ling
et al[6] placed the genus within subtribe Conyzi ̄
naeꎬ whereas Zhang and Bremer[15] treated it as
an isolated genus of Astereae and Bremer[16]
thought it to be a member of subtribe Asterinae
Nesom[9] included it tentatively in subtribe Lag ̄
enophorinae Nesomꎬ which was followed by Ne ̄
som and Robinson[10] Although its genetic sta ̄
tus was supported by Chen et al[14]ꎬ its system ̄
atic position remains uncertain and no molecular
phylogenetic analyses have involved Thespis so
far.
With approximately 10 speciesꎬ Microglossa
DC [14] is distributed in Africa and tropical Asia
Microglossa pyrifolia (Lam ) O Kuntze occurs
widely from Africa to tropical Asiaꎬ and in China
it is a traditional medicine and the only represen ̄
tative of the genus[6ꎬ14] Ling et al [6] assigned
the genus within Conyzinaeꎬ while Zhang and
Bremer[15]and Bremer[16] placed it in the Eriger ̄
on ̄Conyza group Howeverꎬ Nesom[9] placed it
in the Psiadia group of subtribe Baccharidinaeꎬ
and Nesom and Robinson[10] treated it as an un ̄
placed genera in tribe Astereae Thus farꎬ how ̄
everꎬ its phylogeny has not been evaluated by
molecular data.
Based on nuclear ribosomal DNA ( nrDNA)
ITS and ETS ( internal and external transcribed
spacers of 35S ribosomal DNA )ꎬ the present
study aimed to reconstruct the phylogeny of Chi ̄
nese Conyzaꎬ Microglossa and Thespis and to
discuss their classification.
1 Materials and methods
1 1 Taxon sampling and source of sequences
Four Conyza species native to Chinaꎬ three
American Conyza species introduced to Chinaꎬ
Microglossa pyrifolia and Thespis divaricata (Ap ̄
pendix) were sampled in China for sequence
variations in nrDNA ITS and ETS The ITS and
ETS sequences of C sumatrensis (Retz ) Walk ̄
er and C japonica ( Thunb ) Less were pro ̄
duced from previous studies[4]ꎬ and those of the
other seven species were newly generated (Ap ̄
pendix) .
Brouillet et al [2] divided Astereae into six
phylogenetic lineagesꎬ that isꎬ the basal linea ̄
gesꎬ palaeo ̄South American cladeꎬ New Zea ̄
land cladeꎬ Australasian lineagesꎬ South Ame ̄
rican lineages and Northern American lineage
According to Li et al[4]ꎬ there is a seventh cladeꎬ
Bellidinaeꎬ and an eighth cladeꎬ Grangeinae
To determine systematic positions of the three
Chinese generaꎬ all eight phylogenetic lineages
were represented by more or less species in our
ITS matrix and seven phylogenetic lineages in the
combined matrix There were more accessions in
the ITS matrix than in the combined matrix be ̄
cause there was more ITS data available in Gen ̄
Bank[1ꎬ4ꎬ5ꎬ11ꎬ12ꎬ17 ̄26] Strijk[3] and Strijk et al[5] sug ̄
gested that African Conyza species were nested
with two large cladesꎬ so in our ITS matrix some
African Conyza and their relatives represented
two clades (one clade: C ageratoidesꎬ C scabri ̄
daꎬ C pyrrhopappaꎬ C strictaꎬ Psiadia pas ̄
calii Labat and Beentjeꎬ P punctulata (DC )
Vatkeꎻ the other clade: eight Conyza species
and Psiadia argentea Cordem ) in order to reval ̄
uate the relationships among these African plants
and the Chinese taxa In the ITS matrixꎬ two se ̄
quences (HE978363 and GU045830) published
by Strijk et al [5] were recorded in GenBank and
were obtained from the same speciesꎬ Conyza
ulmifolia The two are quite differentꎬ however
Comparison of our ITS tree with that of Strijk et al[3]
showed that HE978363 was the ITS sequence of
C incisa[5] instead of C ulmifolia As a resultꎬ
812 植 物 科 学 学 报 第 32卷
we treated HE978363 as the ITS of C incisa
according to Figures 2 and 3 of Strijk et al [5] The
ITS sequence GU045827 published by Strijk
et al[5] was obtained from C stricta sampled
from eastern Africa We regarded it as C stricta
var stricta distributed in both Asia and Africaꎬ
and was thus distinguished from the other sam ̄
pled varietyꎬ C stricta var pinnatifida Kitam
Conyza stricta var pinnatifida is restricted to
southern Asia and was sampled from Yunnanꎬ
China (Appendix) . Because some taxa of the
ITS matrix were unavailable for ETSꎬ the com ̄
bined ITS / ETS matrix included only 34 ingroup
taxa (Appendix) . In the two matricesꎬ Chrysan ̄
themum coronarium L and Dendranthema indi ̄
cum (L ) Des Moul of tribe Anthemideae and
Calendula officinalis L of tribe Calenduleae (Ap ̄
pendix) were selected as outgroups for the roo ̄
ting of the phylogenetic trees because in molecu ̄
lar phylogenetic analyses[27ꎬ28] Anthemideae and
Astereae are sistersꎬ and Calenduleae is a sister
to tribes Gnaphalieaeꎬ Anthemideae and Astere ̄
ae.
1 2 DNA extractionꎬ polymerase chain reaction
(PCR) and sequencing
Total genomic DNA was isolated from fresh
leaf materials or silica gel ̄dried leaves Amplifi ̄
cation and sequencing were performed using
primers ITS1 and ITS4[29] for the ITS regionꎬ and
using primers Ast ̄8[30] and 18S ̄ETS[31] for the
ETS region.
The PCR mixture contained 1 μL (50-100 ng)
of sample DNAꎬ 2 × 2 μL of primer (10 pmol)ꎬ
5 μL of 10 × PCR bufferꎬ 3 μL of Mg2+(25 mol / L)ꎬ
0 8 μL of deoxyribonucleotide triphosphate (each
25 mol / L )ꎬ 0 5 μL of Taq DNA polymerase
(5 U / mL) and sterile water for a final volume of
50 μL The PCR procedures were as follows: ini ̄
tial denaturation for 4 min at 95℃ followed by
30 cycles of denaturation ( 94℃ꎬ 1 min)ꎬ an ̄
nealing ( 56℃ꎬ 40 s ) and extension ( 72℃ꎬ
1 min)ꎬ and a final extension of 10 min at 72℃.
The PCR fragments were purified with a
UNIQ ̄10 Spin Column PCR Product Purification
Kit (Sangon Biotech Coꎬ Ltdꎬ Shanghaiꎬ Chi ̄
na) following the manufacturers instructions Di ̄
rect sequencing was performed in both directions
by Sangon Biotech Coꎬ Ltd.
1 3 Sequence alignment and phylogenetic ana ̄
lyses
According to our previous study[4]ꎬ bounda ̄
ries of the ITS and ETS regions were determined
All DNA sequences were aligned initially using
Clustal X 1 83[32] and then adjusted manually in
BioEdit[33] The ITS region was analysed sepa ̄
rately and in a combined data set with the ETS
region The incongruence length difference test[34]
was carried out to test the homogeneity between
data sets using PAUP∗ version 4 0b10 with 1000
replicates Maximum parsimony (MP) and Baye ̄
sian inference (BI) were performed for the data
sets using PAUP∗ version 4 0b10[35] and Mr ̄
Bayes version 31 2[36]ꎬ respectively In the MP
analysisꎬ characters were unordered and equally
weightedꎬ gaps were scored as missing dataꎬ
and a heuristic search was implemented with
1000 random additional sequence replicates and
sub ̄tree pruning ̄regrafting branch swapping
Support for clades was estimated by bootstrap
with 1000 replications The MaxTrees setting in
PAUP∗ was set to 5000 for the searches and
bootstrap tests For BI analysis of the ITS region
and combined data setꎬ the best ̄fitting model of
each sequence partition ( ITS1ꎬ ITS2ꎬ 5 8Sꎬ
ETS) was determined using MrModeltest 2 2[37]
The SYM + G model was chosen for the 5 8S re ̄
gionꎬ and the GTR + I + G model for the ITS1ꎬ
ITS2 and ETS regions The Markov chain Monte
Carlo algorithm was run for 1000000 generationsꎬ
resulting in an overall sampling of 10000 trees
912
第 3期
钟彩霞等: 基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究(英文)
The first 3000 trees were discarded as a conser ̄
vation burn ̄inꎬ and the remaining trees were
used to construct the 50% majority rule consen ̄
sus tree.
2 Results
2 1 Characterization of nucleotide data
The ITS data set consisted of 680 aligned nu ̄
cleotidesꎬ of which 383 (56 32%) were variable
and 278 (40 88%) were parsimony ̄informative
The ILD test ( p = 0 123) failed to reject the
combination of the ITS and ETS markers There ̄
foreꎬ a combined analysis of the two regions was
performed using PAUP∗ and MrBayes The com ̄
bined ITS and ETS data set consisted of 1335
aligned nucleotidesꎬ of which 715 (53 56%) were
variable and 495 ( 37 08 %) were parsimony ̄
informative Near the end of the ETS region Eri ̄
geron breviscapus (Vant ) Hand  ̄Mazzꎬ Cony ̄
za canadensis ( L ) Cronqꎬ C bonariensis
(L ) Cronq and C sumatrensis had an insert
as long as 84 bp that was their synapomorphy.
2 2 Phylogenetic analyses
Phylogenetic analyses using ITS and the
combined data sets yielded generally consistent
phylogenetic trees ( Bayesian treesꎻ see Fig 1ꎬ
Fig 2)ꎬ though combined analyses resulted in
more supported clades and more resolved rela ̄
tionships than the independent analysis based on
ITS data In both the ITS tree and combined treeꎬ
the genus Conyza was not monophyletic and oc ̄
curred within three big clades with higher Baye ̄
sian posterior probability ( PP ) and bootstrap
support (BS) (clade 1: PP = 1 00 and BS =
99 in Fig 1ꎬ and PP = 1 00 and BS = 100 in
Fig 2ꎻ clade 2: PP = 1 00 and BS = 68 in
Figs 1 and 2ꎻ clade 7: PP = 0 99 and BS = 90
in Fig 1ꎬ and PP = 1 00 and BS = 100 in
Fig 2) . Furthermoreꎬ in each of the three clades
the Conyza were not grouped in an independent
subclade but mixed with other genera (Fig 1 and
Fig 2) . Microglossa pyrifolia was nested within
clade 2 and was not closely related to most Chi ̄
nese Conyza (Fig 1 and Fig 2)ꎬ whereas Thes ̄
pis divaricata belonged to clade 7 with most Afri ̄
can and Chinese Conyza sampled.
3 Discussion
3 1 Phylogenetics of Chinese Conyza and the
taxonomic implications
Three American Conyza speciesꎬ C bona ̄
riensisꎬ C canadensis and C sumatrensisꎬ are
widely distributed throughout China They were
found to be nested in the North American clade
with high supported values (clade 1: PP = 1 00
and BS = 99 in Fig 1ꎻ PP = 1 00 and BS = 100
in Fig 2) of Astereae and closely related to Eri ̄
geron In particularꎬ Erigeron breviscapus shared
an 84 bp segment of ETSꎬ a synapomorphic
characterꎬ with C canadensisꎬ C bonariensis
and C sumatrensis These results support previ ̄
ous reports[1 ̄4ꎬ7ꎬ11 ̄13] and the taxonomic treatment
of Chen et al[14] that these three Conyza were
placed in the genus Erigeron Although previous
studies also showed that New World Conyza were
not closely related to Old World Conyzaꎬ only
one Asian speciesꎬ C japonica (Eschenbachia
japanica)ꎬ was included[4] Our study showed
that in the trees (Fig 1 and Fig 2) the three in ̄
vasive alien Conyza species were far from the
Conyza native to China.
Our phylogenetic trees ( Fig 1 and Fig 2)
demonstrated that Chinese Conyza were biphyle ̄
tic because Conyza stricta var stricta and var
pinnatifida belonged to clade 2 and were not
close to clade 7 where the other three Chinese
Conyza occurred (Fig 1 and Fig 2) . Conyza in ̄
cisa and the two C stricta varieties occurred in a
well ̄supported sub ̄clade (PP = 1 00ꎬ BS = 95ꎻ
Fig 1) . Unexpectedlyꎬ var pinnatifida initially
022 植 物 科 学 学 报 第 32卷
Aster verticillatus
Aster tataricus
Aster ageratoides lasiocladus
Sheareria nana
Baccharis neglecta
Aster lavanduliifolius
Calotis hispidula
Lagenophora pumila
Myriactis nepalensis
Myriactis wightii
Conyza chilensis
Conyza sumatrensis
Conyza bonariensis
Conyza canadensis
Erigeron breviscapus
Symphyotrichum subulatum
Eurybia sibirica
Bellis perennis
Galatella dahurica
Tripolium vulgare
Dichrocephala auriculata
Nidorella polycephala
Grangea maderaspatana
Conyza stricta stricta
Conyza stricta pinnatifida
Conyza incisa
Welwitschiella nereifolia
Conyza ageratoides
Psiadia punctulata
Microglossa pyrifolia
Psiadia pascalii
Conyza scabrida
Conyza pyrrhopappa
Psiadia argentea
Conyza attenuata
Conyza pinnata
Conyza limosa
Conyza subscaposa
Conyza muliensis
Conyza japonica
Conyza gouanii
Conyza tigrensis
Conyza aegyptiaca
Conyza blinii
Conyza ulmifolia
Thespis divericata
Madagaster madagascariensis
Oritrophium peruvianum
Felicia aethiopica
Felicia filifolia
Mairia hirsuta
Pleurophyllum hookeri
Celmisia mackaui
Llerasia macrocephala
Chiliotrichum diffusum
Nannoglottis delavayi
Printzia polifolia
Chrysanthemum coronarium
Dendranthema indicum
Calendula officinalis
var.
var.
var.
AIS = Astereae incertae sedis
AL = Australasian lineages
BE = Bellidinae
BL = early-branching lineages
BS = Eschenbachia
GR = Grangeinae
NA = North American lineage
NZ = New Zealand clade
OG = outgroup
PSA = palaeo South American clade
SA = South American lineages
1.00/96
1.00/100 AL
SA
AL
NA
BE
GR
AIS
ES
PSA
BL
NZ
PSA
BL
OG
0.99/-
1.00/100
1.00/781.00/100
1.00/99
1.00/87
0.96/-
1.00/88
1.00/-
1.00/54
0.98/64
1.00/68
1.00/-
3
1.00/95
2
0.99/53
0.94/-
4
1.00/100
5
1.00/100
1.00/-
6
0.97/77
1.00/90
0.99/96
0.97/-
0.97/62
-/78
0.99/90
7
0.98/51
0.99/67
0.93/54
0.98/95
0.99/98
1.00/99
0.94/99
1.00/100
1
Bayesian posterior probabilities ( ≥ 0 90) and bootstrap values ( ≥ 50%) are indicated above the branchesꎻ “-” or no number indi ̄
cates that Bayesian posterior probabilities are < 0 90 and or bootstrap percentages are < 50% For convenience of discussion and reada ̄
bilityꎬ some clades are indicated by numbers below the branch Abbreviations of the lineages are shown on the right side of the taxa and
explained (see frame at the top left ̄hand side of the figure) .
Fig 1 50% majority rule consensus tree from Bayesian analysis of nuclear ribosomal DNA internal
transcribed spacer sequences
122
第 3期
钟彩霞等: 基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究(英文)
Aster verticillatus
Aster tataricus
Aster ageratoides lasiocladus
Sheareria nana
Lagenophora pumila
Aster lavanduliifolius
Myriactis nepalensis
Myriactis wightii
Conyza sumatrensis
Conyza bonariensis
Conyza canadensis
Erigeron breviscapus
Symphyotrichum subulatum
Eurybia sibirica
Bellis perennis
Galatella dahurica
Tripolium vulgare
Dichrocephala auriculata
Nidorella polycephala
Grangea maderaspatana
Conyza stricta pinnatifida
Microglossa pyrifolia
Conyza muliensis
Conyza japonica
Conyza blinii
Thespis divericata
Madagaster madagascariensis
Felicia aethiopica
Felicia filifolia
Oritrophium peruvianum
Pleurophyllum hookeri
Celmisia mackaui
Llerasia macrocephala
Chiliotrichum diffusum
Chrysanthemum coronarium
Dendranthema indicum
Calendula officinalis
var.
var.
1.00 96/
1.00 100/
1.00/100
1.00 100/
1.00 -/
1.00 98/
1.00 100/
1.00 100/
1.00 87/
1.00 98/
0.98/70
1.00/68 1.00/66
1.00/98
1.00/67
-/76
1.00/100
1.00/100
1
0.99 -/
1.00 100/
2 3
7
1.00/95
0.99/79
0.99 -/
1.00/62
1.00 100/
1.00 -/
ES
AL
NA
BE
GR
AIS
BL
PSA
NZ
PSA
OG
Bayesian posterior probabilities ( ≥ 0 90) and bootstrap values ( ≥ 50%) are indicated above the branchesꎻ “-” or no number in ̄
dicates that Bayesian posterior probabilities are < 0 90 and or bootstrap percentages are < 50% Some clades are indicated by num ̄
bers below the branchꎬ identical to Fig 1 Abbreviations of the lineages are shown on the right side of the taxa and are identical to
those in Fig 1.
Fig 2 50% majority rule consensus tree from Bayesian analysis of the combined data set
(nuclear ribosomal DNA internal and external transcribed spacer sequences)
combined with C incisa rather than with var
stricta (Fig 1)ꎬ although the combination of the
two taxa was lowly supportedꎬ implying that C
stricta might not be monophyletic and is in need
of a taxonomic revision Although C stricta and
C incisa were embedded within the same large
clade (clade 2ꎻ Fig 1) as the other three Afri ̄
can Conyza (C ageratoidesꎬ C scabrida and
C pyrrhopappa)ꎬ the former and latter did not
form a monophyletic cladeꎬ indicating that the
former should be treated as a separate genus
Furthermoreꎬ the new genus belonged to sub ̄
tribe Grangeinae because it was embedded within
clade 2 (PP = 100 and BS = 68 in Fig 1 and
Fig 2)ꎬ i e subtribe Grangeinaeꎬ which includ ̄
ed typical representatives such as Grangeaꎬ Di ̄
chrocephalaꎬ Nidorella[6ꎬ10] and Welwitchiella [1] .
In the ITS treeꎬ the other three Chinese
Conyzaꎬ C bliniiꎬ C muliensis and C japoni ̄
caꎬ formed the highly supported clade 7 (PP =
0 99 and BS = 90 in Fig 1) with seven African
Conyza species and one African ̄Asian Conyza
species (C aeagyptiaca) . The combined tree
(Fig 2) indicated that these three Chinese Cony ̄
za belonged to the same clade (clade 7ꎻ PP =
1 00ꎬ BS = 100) . Chen et al [14] resurrected
Eschenbachia typified by E globosa Moenchꎬ
a synonym of Eschenbachia aegyptiaca ( L )
222 植 物 科 学 学 报 第 32卷
Brouillet ( = Conyza aegyptiaca (L ) Aiton) and
delimited Eschenbachia to include Chinese native
Conyza (excluding C stricta) . Our phylogenetic
analyses showed that clade 7 (Fig 1 and Fig 2)
included most sampled taxa of Chinese and Afri ̄
can Conyza and corresponded to the genus Es ̄
chenbachia.
The African origin of Eschenbachia is based
on two reasons Firstꎬ the diversity center of the
genus was located in Africa where most Eschen ̄
bachia species occur ( see clade 7 in Fig 1) .
Secondlyꎬ it seems that Eschenbachia might
originate from African taxa because clade 7 was
a sister to the African Madagaster madagasca ̄
riensis clade (Fig 1) and was derived from the
African basal lineages (BLꎻ Fig 1 and Fig 2) . If
the African origin of Eschenbachia is trueꎬ Es ̄
chenbachia might have arrived in southern China
by long ̄distance dispersal from Africa Trans ̄
continental origins have been well documented
within Astereae[11ꎬ38] .
Furthermoreꎬ the dispersal happened sever ̄
al times because the four sampled Chinese Es ̄
chenbachia speciesꎬ including Thespis divarica ̄
ta ( discussed below)ꎬ did not form a single
clade ( Fig 1) . For exampleꎬ C japonica was
more closely related to African Conyza than C
muliensis and C blinii endemic to Chinaꎬ and
the two endemic species seem phylogenetically
closer to African Conyza than to each other.
Nesom[9] and Nesom and Robinson[10] sug ̄
gested that African Conyza were members of
subtribe Grangeinae or closely related to genera
of the sbutribeꎬ which was supported by Brouillet
et al [1] but not fully by our results ( Fig 1 and
Fig 2) . Although a few African and Asian Conyza
belonged to Grangeinaeꎬ most belonged to Es ̄
chenbachia and were not closely related to
Grangeinae ( Fig 1 and Fig 2 ) . In the trees
(Fig 1 and Fig 2)ꎬ clade 2 (subtribe Grangein ̄
ae) belonged to a big clade with subtribe Belli ̄
nae ( represented by Bellis perennis )ꎬ North
American and Australasian lineagesꎬ whereas
clade 7 ( Eschenbachia) was sister to the big
clade but not within it It isꎬ thereforeꎬ reasona ̄
ble to treat clade 7 (Eschenbachia) as a new
separate subtribe because Eschenbachia can not
be placed within any existent subtribes.
3 2 Phylogenetics of Thespis and the taxonomic
implications
Up to nowꎬ botanists have recognized the
generic status of Thespisꎬ but placed it in diffe ̄
rent subtribesꎬ Conyzinae[6]ꎬ Asterinae[16] or
Lagenophorinae[9ꎬ10]ꎬ or treated it as an isolated
genus of Astereae[15]ꎬ or avoided discussing its
subtribal position[14] Howeverꎬ our phylogenetic
trees ( Fig 1 and Fig 2 ) rejected all previous
treatments Thespis belonged to the strongly
supported clade 7 (PP = 0 99 and BS = 90 in
Fig 1ꎻ PP = 1 00 and BS = 100 in Fig 2) and
was nested among some African and Chinese
Conyza (Eschenbachia)ꎬ indicating that Thespis
should be merged into Eschenbachia Thespis
species are annual herbs with alternate simple
leavesꎬ small disciform capitula with 2 ̄seriate
herbaceous subequal phyllariesꎬ multiseriate fer ̄
tile pistillate floretsꎬ and have slightly com ̄
pressed achenes with 1 ̄seriate barbellate bristle
pappus that is largely identical to that in other
species of Eschenbachia[14] . Thespis speciesꎬ
howeverꎬ are quite different from other Eschen ̄
bachia species in some important characteris ̄
tics Firstlyꎬ the heads in Thespis are fascicled
in divaricately branched corymbiform synflores ̄
cences Secondlyꎬ their disc florets are function ̄
ally male and sterile Finallyꎬ the disc florets
sometimes have no pappus and the peripheral
female florets lack corolla Thereforeꎬ Thespis
would be better treated as a section of Eschen ̄
bachia.
322
第 3期
钟彩霞等: 基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究(英文)
3 3 Phylogenetics of Microglossa and the taxo ̄
nomic implications
Ling et al [6]ꎬ Zhang and Bremer[15] and
Bremer[16] suggested that Microglossa was relat ̄
ed to Conyzaꎬ whereas Nesom and Robinson[10]
could not assign it to any known subtribe of As ̄
tereae The two trees ( Figs 1 and 2) demon ̄
strated that M pyrifolia was embedded deeply in
clade 2 (subtribe Grangeinae)ꎬ which does not
support any previous studies on the subtribal po ̄
sition of the genus Microglossa pyrifolia formed
a strongly supported clade (PP = 1 00 and
BS = 100 in Fig 1) with Psiadia pascalii and fur ̄
ther combined C scabrida and C pyrrhopappa
to form clade 6 (PP = 1 00ꎻ Fig 1) . Clade 6
was one of three subclades in clade 4ꎬ as was
clade 5 (PP = 1 00 and BS = 100 in Fig 1)ꎬ
the principal part of Psiadia re ̄delimitated by Stri ̄
jk[3] and Strijk et al [5] Strijk[3] expanded Psia ̄
dia to include some African Conyza such as C
scabrida and C pyrrhopappa of clade 6 (Fig 1).
The latter two species were not embedded within
clade 5 ( Fig 1)ꎬ however Strijk[3] transferred
C scabrida and C pyrrhopappa to Psiadiaꎬ
which indicated that clade 4 could be the genus
Psiadia and that Welwitschiella and Microglossa
(at least M pyrifolia) should be merged into Psi ̄
adia Microglossaꎬ howeverꎬ is quite different
from Psiadia by its inflorescences of dense cor ̄
ymbose to subglobose cymes ( vs corymbose
ones in Psiadia)ꎬ receptacles with no paleae (vs
minutely paleae)ꎬ white ( vs yellow ) marginal
and disc floretsꎬ and perfect ( vs functionally
male) disc florets Alsoꎬ Welwitschiella differs
from Psiadia by its herbaceous ( vs shrubby )
stemsꎬ receptacles with no paleae ( vs minutely
paleate) and pappus of toothed scales (vs bris ̄
tles) . Thereforeꎬ it is reasonable to maintain the
generic status of Welwitschiella and Microglossa
and treat Psiadia pascaliiꎬ C scabrida and C
pyrrhopappa as members of Microglossa rather
than as Psiadia species More evidence from
molecular dataꎬ cytology and morphology is nec ̄
essary to determine the taxonomic status of Mi ̄
croglossaꎬ Welwitschiellaꎬ Psiadia pascaliiꎬ C
scabrida and C pyrrhopappa.
Acknowledgements: We thank the referees for their
useful commentsꎬ suggestions and questionsꎻ Dr
YANG Fu ̄Sheng for his help in analysing the molecular
data and Ms ZHANG Ping for field assistance.
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歧伞菊属(菊科紫菀族)分子系统学研究(英文)
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Appendix: Speciesꎬ sourcesꎬ voucher numbers and GenBank accessions ( ITSꎬ ETS) used in this
study Information of the sources and voucher numbers is limited to those species in which sequences
were newly generated Some species were unavailable for ETS (★: Data are being submitted to NCBI).
Aster ageratoides Turcz var lasiocladus (Hayata) Hand  ̄Mazzꎬ JN543781ꎬ JN543782[4] ꎻ Aster lavanduliifolius
Hand  ̄Mazzꎬ JN543781ꎬ JN543782[4] ꎻ Aster tataricus L fꎬ JN543748ꎬ JN543749[4] ꎻ Aster verticillatus ( Rein ̄
wardt) Brouilletꎬ Semple and Y L Chenꎬ JN543706ꎬ JN543707[4] ꎻ Baccharis neglecta Britton U97604[17] ꎻ Bellis pe ̄
rennis Lꎬ JN315918ꎬ JN315942[4] ꎻ Calendula officinalis Lꎬ JN315941ꎬ JN315965[4] ꎻ Calotis hispidula (F Muell )
F Muellꎬ AB196597[23] ꎻ Celmisia mackaui Raoulꎬ AF422115[20] ꎬ HQ439825[25] ꎻ Chiliotrichum diffusum (G Forst )
Kuntze AF046945[11] ꎬ DQ479128[22] ꎻ Chrysanthemum coronarium Lꎬ JN315939ꎬ JN315963 [4] ꎻ Conyza aegyptiaca
(L ) Aitꎬ GU045821 [5] ꎻ Conyza ageratoides DCꎬ GU045822[5] ꎻ Conyza attenuata DCꎬ GU045823[5] ꎻ Conyza bli ̄
nii H Lévꎬ Chinaꎬ Kunmingꎬ lwp 0708282 (HNNU)ꎬ ★ꎬ★ꎻ Conyza bonariensis (L ) Cronqꎬ Chinaꎬ Changshaꎬ
lwp 1103005 (HNNU)ꎬ ★ꎬ★ꎻ Conyza canadensis (L ) Cronqꎬ Chinaꎬ Changshaꎬ lwp 1103009 (HNNU)ꎬ ★ꎬ★ꎻ
Conyza chilensis Sprengꎬ AF118510[12] ꎻ Conyza gouanii ( L ) Willdꎬ AF046948[11] ꎻ Conyza incisa Aitꎬ
HE978363[5] ꎻ Conyza japonica (Thunb ) Lessꎬ JN315938ꎬ JN315962[4] ꎻ Conyza limosa O Hoffmꎬ GU045824[5] ꎻ
Conyza muliensis Y L Chenꎬ Chinaꎬ Sichuanꎬ lwp 0708074 ( HNNU)ꎬ ★ꎬ★ꎻ Conyza pinnata ( L f ) Kuntzeꎬ
GU045825[5] ꎻ Conyza pyrrhopappa Sch Bip ex A Richꎬ AF046953[11] ꎻ Conyza scabrida DCꎬ GU045826[5] ꎻ
Conyza stricta Willd var pinnatifida (D Don) Kitamꎬ Chinaꎬ Kunmingꎬ lwp 1009012 (HNNU)ꎬ ★ꎬ★ꎻ Conyza stricta
Willd var strictaꎬ GU045826[5] ꎻ Conyza subscaposa O Hoffmꎬ GU045828[5] ꎻ Conyza sumatrensis (Retz ) Walkerꎬ
JN315923ꎬ JN315947[4] ꎻ Conyza tigrensis Oliv and Hiernꎬ GU045829[5] ꎻ Conyza ulmifolia ( Burm f ) Kuntzeꎬ
622 植 物 科 学 学 报 第 32卷
GU045830[5] ꎻ Dendranthema indicum (L ) Des Moulꎬ JN315940ꎬ JN315964[4] ꎻ Dichrocephala auriculata (Thunb )
Druceꎬ JN315919ꎬ JN315943[4] ꎻ Erigeron breviscapus ( Vant ) Hand  ̄Mazzꎬ JN315925ꎬ JN315949[4] ꎻ Eurybia
sibirica (L ) Nesomꎬ AY772421ꎬ AY772435[21] ꎻ Felicia aethiopica (Burm f ) Bolus and Wolley ̄Dod ex Adamson
and T M Salterꎬ DQ478997ꎬ DQ479054[24] ꎻ Felicia filifolia (Vent ) Burtt Davyꎬ FJ457937[1] ꎬ AF319703[18] ꎻ Gala ̄
tella dahurica DCꎬ JN315935ꎬ JN315959[4] ꎻ Grangea maderaspatana (L ) Poirꎬ JN315920ꎬ JN315944[4] ꎻ Lageno ̄
phora pumila ( G Forst ) Cheesemanꎬ DQ479037ꎻ DQ479094[24] ꎻ Llerasia macrocephala ( Rusby ) Pruskiꎬ
JQ042751ꎬ JQ042790[26] ꎻ Madagaster madagascariensis (Humbert) Nesomꎬ DQ479031ꎬ DQ479087[24] ꎻ Mairia hir ̄
suta DCꎬ FJ457929[1] ꎻ Microglossa pyrifolia ( Lam ) O Kuntzeꎬ Chinaꎬ Yunnanꎬ lwp 0804078 (HNNU)ꎬ ★ꎬ★ꎻ
Myriactis nepalensis Lessꎬ JN315921ꎬ JN315945[4] ꎻ Myriactis wightii DCꎬ JN315922ꎬ JN315946[4] ꎻ Nannoglottis
delavayi (Franch ) Ling and Chenꎬ AY017167[19]ꎻ Nidorella polycephala DCꎬ DQ478999ꎬ DQ479057[24]ꎻ Oritrophium
peruvianum ( Lam ) Cuatrꎬ DQ479117[24]ꎬ JQ042796[26]ꎻ Pleurophyllum hookeri Buchꎬ HQ439864ꎬ HQ439853[25]ꎻ
Printzia polifolia ( L ) Hutchꎬ FJ457927[1] ꎻ Psiadia argentea Cordemꎬ HE978394[5] ꎻ Psiadia pascalii Labat and
Beentjeꎬ HE978450[5] ꎻ Psiadia punctulata (DC ) Oliv and Hiern ex Vatkeꎬ HE978457[5] ꎻ Sheareria nana S Mooreꎬ
JN543703ꎬ JN543704[4] ꎻ Symphyotrichum subulatum (Michx ) Nesomꎬ JN315927ꎬ JN315951[4] ꎻ Thespis divaricata
DCꎬ Chinaꎬ Yunnanꎬ lwp 0804067 ( HNNU )ꎬ ★ꎬ★ꎻ Tripolium vulgare Neesꎬ JN315937ꎬ JN315961[4] ꎻ Wel ̄
witschiella nereifolia O Hoffmꎬ FJ457943[1] .
(责任编辑: 张 平)
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第 3期
钟彩霞等: 基于 2个核糖体 DNA序列的国产白酒草属、 小舌菊属和
歧伞菊属(菊科紫菀族)分子系统学研究(英文)