全 文 ：·快讯 Short Communication·
从 ITS序列探讨青藏高原特有植物华福花属的亲缘关系
刘建全1 ,2　陈之端2　路安民2
(1.中国科学院西北高原生物研究所 , 西宁 810001;2.中国科学院植物研究所系统与进化植物学开放研究实验室 , 北京 100093)
摘要:　对青藏高原特有的濒危植物华福花(Sinadoxa corydalifolia C.Y.Wu , Z.L.Wu et R.F.Huang)的核糖体
DNA中的内转录间隔区(ITS)序列及 5.8S rRNA基因的序列进行了测定。同川续断目和五加目有关类群序列的比
较及分支分析表明华福花属与五福花属(Adoxa L.)近缘 ,不支持它可能与五加目或败酱科有亲缘关系的假设。尽
管形态上它与五福花属分化十分明显 ,但 ITS 碱基分异却较小。引起其系统位置发生争论的外部形态为什么进化
得如此之快 ,是值得进一步深入探讨的课题。
关键词:　华福花;内转录间隔区(ITS);系统学;进化
中图分类号:Q941　　　文献标识码:A　　　文章编号:0577-7496(2000)06-0656-03
The Phylogenetic Relationships of an Endemic Genus Sinadoxa in the Qinghai_
Xizang Plateau:Evidence from ITS Sequence Analysis
LIU Jian_Quan1 , 2 , CHEN Zhi_Duan2 , LU An_Ming2
(1.Northwest Plateau Institute of Biology , The Chinese Academy of Sciences , Xining 810001 , China;
2.Laboratory of Systematic and Evolutionary Botany , Institute of Botany , The Chinese Academy of Sciences , Beijing 100093, China)
Key words:　Sinadoxa corydalifolia;internal transcribed spacers(ITS);systematics;evolution
　　The monotypic genus , Sinadoxa , was recently es-
tablished by Wu et al
[ 1] .The only species , Sinadoxa
corydalifolia , was found in the several localities of the
southern Qinghai Province in China during the 1960s and
the 1970s.However , in our recent field expeditions from
1995 to 1999 , we found that most populations of the
species had disappeared.This species has become one of
the exceedingly endangered species in the Qinghai_Xizang
Plateau.Because of the difficulty of obtaining the materi-
als , Sinadoxa was poorly studied.Although it is morpho-
logically different from Adoxa and Tetradoxa in the char-
acters of rhizome , leaf , calyx and ovary , especially in the
highly complicated inflorescence like a spike with several
glomerate interrupted clusters , most Chinese researchers
accepted Sinadoxa as a natural member of the Adoxaceae
s.s., to be closely related to Adoxa[ 2-3] .However ,
Cronquist[ 4] thought that the gynoecium of Sinadoxa is
very suggestive of the Valeraceae.Donoghue et al[ 5]
pointed out that “Sinadoxa differs in many respects and
may be more closely related to Araliales than to Adoxa” .
Although later Donoghue① recognized that it might be
more reasonable to treat Sinadoxa within the Adoxa clade
of the Dipsacales , he further declared that this has not
been adequately tested.The karyomorphology of Sinadoxa
strongly indicated its close relationship with Adoxa
[ 6] .
The present paper describes the study of the phylogenetic
relationships of Sinadoxa using ITS sequence analysis.
1　Materials and Methods
1.1　Plant materials
Leaves of S . corydalifolia were collected in
Nangchian county of the southern Qinghai(alt.3 900 m)
and dried in silica gel in June , 1999.Voucher specimens
(Liu Jianquan 625)were deposited in HNWP.Sequences
of Adoxa , Virbunum , Sambucus and other taxa of Dip-
sacales and Araliales were downloaded from the GenBank
(also see Eriksson and Donoghue[ 7]).Their accession
numbers are U88552_U88557 , U88194 , U88198 ,
U41381_U41382 and AF077875.
1.2　DNA sequencing
Total DNA was extracted as described by He et
al[ 8] .PCR reactions were carried out following Zhang et
al[ 9] and Wang et al[ 10] and double_stranded DNA was
cycle_sequenced using PCR systems 9 600 kits and ana-
lyzed on an ABI 377 automated sequencer from both di-
rections.
＊　Received:2000-01-25　Accepted:2000-03-30
Foundation item:A grant f rom President Foundation of the Chinese Academy of Sciences;The National Natural Science Foundation of China(39930020).
① Donoghue M J.Phylogeny and phylogenetic taxonomy of Dipsacales.Amer J Bot , 1995 , 82(Abstracts):108.
植　物　学　报　2000 , 42(6):656-658 　 　 　 　 　 　 　
Acta Botanica Sinica
Table 1　Pairwise genetic distance(below:absolute distances/ above:mean distances)of nrDNA ITS and 5.8 s rDNA sequences of Sinadoxa
and other 7 species of Adoxa , Sambucus , and Viburnum
1 2 3 4 5 6 7 8
　1 Sambucus callicarpa - 0.031 0.034 0.138 0.132 0.242 0.215 0.195
　2 Sambucus gaudichaudiana 19 - 0.003 0.142 0.132 0.239 0.220 0.188
　3 Sambucus australasica 21 2 - 0.143 0.133 0.242 0.221 0.189
　4 Sinadoxa corydalifolia 84 86 87 - 0.034 0.269 0.235 0.231
　5 Adoxa moschatellina 80 80 81 21 - 0.259 0.236 0.222
　6 Viburnum sieboldii 112 114 115 122 122 132 - 0.084
　7 Viburnum lentago 103 99 100 122 117 119 44 -
　8 Viburnum dentatum 106 107 108 125 119 122 33 34
Fig.1.　Single and most parsimonious tree for Sinadoxa and related taxa based on nrDNA ITS and 5.8S rDNA sequences using three Vibur-
num species as the outgroup.
The tree has 239 steps , with CI=0.908 and RI=0.908.Base substitutions are indicated above branches.Bootstrap values are indicated be-
low branches.
1.3　Data analysis
The sequences of S .corydalifolia and those from
Araliales and Dipsacales were aligned and compared using
CLUSTALV and the phylogenetic analysis was conducted
using PAUP 3.1.1 following Wang et al[ 10] .
2　Result and Discussion
The sequences of the nuclear ribosomal DNA ITS re-
gion and 5.8S rRNA gene of Sinadoxa corydalifolia are
611 bp (deposited in the GenBank data base , accession
no.AF248611).The length of ITS1 is 225 bp , 5.8S
rRNA is 164 bp and ITS2 is 222 bp.We compared the
sequences of S .corydalifolia with those from Araliales
and Dipsacales and found that the sequences of S .cory-
dalifolia are similar to those of Adoxa , Viburnum and
Sambucus , but distinctly different from other taxa of these
two orders.In the first unrooted analysis using all ITS da-
ta of these orders , Sinadoxa is always combined with the
Adoxa supported by the bootstrap value of 100%.But in
such an analysis , there appeared many unresolved clades
among the representative taxa of the two orders , which
might be due to the long branch attraction and the homo-
plasy evolution of ITS sequences at the higher taxonomic
level.
Adoxa , Viburnum and Sambucus comprise a natural
group , and were sometimes treated together as the ex-
panded Adoxaceae[ 11 ,12] .The floral ontogenetical study
further indicated that Adoxa is closely related to Sambu-
cus[ 16] .Therefore , Adoxa and Sambucus were together
treated as the Adoxoideae while Viburnum comprises the
other subfamily of Adoxaceae s.l.[ 11-13] .Eriksson and
Donoghue[ 7] discussed the relationships of Adoxa and
Sambucus using Viburnum as the outgroup based on the
ITS and morphological data.All Sambucus species formed
a natural clade , and Adoxa comprised the other clade.In
the last analysis of ITS data presented here in Table 1 and
6 期 刘建全等:从 ITS 序列探讨青藏高原特有植物华福花属的亲缘关系(英) 657　
Fig 1., we mainly used the data of these three genera to
be compared to that of Sinadoxa.Six species are further
selected to represent respectively Sambucus clade and
Viburnum clade.
Using the Viburnum as the outgroup , the heuristic ,
exhaustive and bound_branch searches all resulted in only
one tree(Fig.1).Sinadoxa is the sister group of Adoxa
supported by the bootstrap value of 100%and 46 common
base substitutions.The absolute distance between them is
only 21 bp and the relative distance is only 0.034 , which
is the same as that between Sambucus callicarpa and S .
australasica , two representatives from two sections of the
genus.The branch length or substitutions from the com-
mon node to Adoxa is 8 , while the length to Sinadoxa is
13 , only four more steps.This result is surprising and in-
teresting , because two genera are so morphologically dif-
ferent.The habits of two genera are also distinctive.
Sinadoxa grows on the alpine area at elevations between
3900-4800 m while Adoxa grows under woods at alti-
tudes between 2300-3600 m.But why its gross_morphol-
ogy evolves so fast that causes the dispute of the systemat-
ic position of the genus while the sequence divergence is
so low , needs a further study.Another pending work is to
study the endangering mechanisms and to find the proper
protection strategies of S.corydalifolia , because the in-
dividuals of this species decreased annually.At any rate ,
the present investigation supports the close relationship of
Sinadoxa and Adoxa , which is consistent with our previ-
ous karyomorphological study that Sinadoxa has the same
basic chromosome number and similar karyotype with
Adoxa
[ 6] .
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