全 文 ：Paeonia anomala的核型研究
潘 锦1 , 3 , 张大明1
?
, 王 超1 , 桑 涛2 , 潘开玉1 , 洪德元1
( 1 中国科学院植物研究所系统与进化植物学国家重点实验室 , 北京 100093 ; 2 Department of Plant Biology,
Michigan State University, East Lansing, MI 48824 , USA ; 3 中国科学院研究生院 , 北京 100049)
摘要 : 报道了 Paeonia anomala L . 的核型 , 澄清了新疆阿尔泰地区分布的该物种的染色体数目。分布于该
地区的 Paeonia anomala L . 的核型组成 : 2n= 2x= 10 = 6m+ 2sm+ 2st。该类群核型与该属其它类群一致———
2A 型。在综合比较分析该属染色体参数以及核型不均一性参数 , 包括最长?最短染色体比 (L1?Ln ) 和染色
体不对称系数 ( CKOA ) 的基础上 , 我们发现该属三个组在核型上没有明显分化 , 仅在木本类群
(Sect. Moutan DC .) 和草本类群 (Sect. Onaepia Lindley和 Sect. Paeonia) 之间存在微小差异。此外 , 作为二倍
体类群 , 新疆阿尔泰地区分布的 Paeonia anomala L . 很可能是二倍体杂种 , 这将为研究被子植物的父母本
同倍化杂交式物种起源提供一个很好的研究材料。
关键词 : Paeonia; Paeonia anomala; 核型 ; 染色体不对称系数
中图分类号 : Q 943 文献标识码 : A 文章编号 : 0253 - 2700(2006)05 - 488 - 05
Karyotypic Study on Paeonia anomala (Paeoniaceae)
PAN J in1 , 3 , ZHANG Da-Ming1 * , WANG Chao1 , SANG Tao2 ,
PAN Kai-Yu
1
, HONG De-Yuan
1
( 1 State Key Laboratory of Systematic and Evolutionary Botany, Instituteof Botany, Chinese Academy of Sciences, Beijing
100093 , China; 2 Department of Plant Biology, Michigan StateUniversity, East Lansing, MI 48824 , USA ;
3 GraduateSchool , Chinese Academy of Sciences, Beijing 100039 , China)
Abstract : This is thefirst report of thechromosome karyotypeof P . anomala fromthe Altai populations . Thepopulations
sampled from the Altai region of Xinjiang, China, are diploid ( 2n= 10) , with the 2A type karyotype and the karyotypic
formula of 2n= 2x= 10 = 6m+ 2sm+ 2st . Based on karyotypic and asymmetric parameters, i . e . , meanarm ratio, ra-
tio of longest?shortest chromosomes (L1?Ln ) , coefficient of karyotypic overall asymmetry (CKOA) , itwas foundthat there
wasno apparent differentiation in karyotype features among all the three sections of the genus Paeonia, although some
slight karyotypic differences were found between the woody section ( Sect. Moutan DC .) and the herbaceous sections
(Sect. Onaepia Lindley and Sect. Paeonia) . The diploid status of the P . anomala populations, revealed in this study,
implies that the species, assumed as interspecific hybridization origin, may add to the handful examples of homoploid hy-
brid speciation in angiosperms .
Key words: Paeonia; Paeonia anomala; Karyotype; Coefficient of Karyotypic Overall Asymmetry ( CKOA)
The genus Paeonia L ., with chromosomes large
in size but low in number, has attracted considerable
cytologists′and cytogenetists′attention since the 1930′
s . The genus is divided into three sections, Moutan
DC ., Onaepia Lindley and Paeonia, with a total of
ca . 30 species (Hong et al , 2001 ) distributed in the
Northern Hemisphere . Among the species cytologically
studied, all of those in sects. Moutan and Onaepia are
云 南 植 物 研 究 2006 , 28 (5) : 488～492
Acta Botanica Yunnanica

? ?Corresponding author . E - mail: zhangdm@ ibcas. ac. cn
Received date: 2005 - 11 - 21 , Accepted date: 2006 - 04 - 10
作者简介 : 潘锦 ( 1977 - ) 女 , 在读博士研究生 , 主要从事系统与进化植物学研究。
diploid ( 2n = 10 , Dark, 1936; Stebbins et al ,
1938 and 1939; Stern, 1946; Delay, 1947; Ga-
jewski, 1948; Okada et al , 1979; Li , 1982 ) ,
while those in Sect. Paeonia are either diploidor tetra-
ploid ( 2n= 20) (Dark, 1936; Stern, 1946; Sopo-
va, 1971; Hong et al , 1988 ) , except two species
where both diploid and tetraploid were found intraspe-
cifically (Sopova, 1971; Hong et al , 1988 ) . How-
ever, due to lack of a universal parameter to measure
thekaryotypes, not all species reported have been suf-
ficiently analyzed karyotypically so far . Among the
well-analysed species the comparisons were made most-
ly between diploids and tetraploids, or between two
sections ( Tzanoudakis, 1983; Punina, 1987; Hong
et al , 1988; Zhang and Hou, 1996) .
Paeonia anomala L . is distributed in Xinjiangof
northwestern China, northeastern Kazakhstan, and
northern Mongolia and eastern of Russia (Hong et al ,
2001) . The accessions from northern Xinjiang were
once misidentified as P . sinjiangensis Pan ( Pan,
1979) but later were replaced ( Hong, 2004) . In the
molecular systematic studies, Sang and Zhang (1999)
speculated that the populations of P . anomala ( under
the name of P . sinjiangensis) distributed along Altai
Mountains, Xinjiang, were interspecific hybrids . Be-
cause the Altai populations showed heterozygosity at
nuclear genes, Adh1 and Adh2 , andwerewell isolat-
ed with a sympatric diploid species, P . intermedia
(Hong et al , 1994 ) , they suggested that the Altai
populations of P . anomala ( P . sinjiangensis) were
possibly allotetraploid, although the ploidy level Xin-
jiang′s populationsof P . anomalawas thennot known .
This paper is to report the chromosomal number and the
karyotypeof the Altai populations of P . anomala .
Materials and Methods
Two populations of Paeonia anomala L . involved in this
study were collected fromAltai District, Xinjiang, China . One
population (XJ021 and XJ027) was collectedfromHanasi Nature
Reserve (喀纳斯自然保护区 ) , Buerjin County (布尔津
县) , and another (XJ034 ) fromBaihaba HolidayVillage (白哈
巴渡假村 ) , Habahe County ( 哈巴河县 ) . All the vouchers
are preserved in the Herbarium, Institute of Botany, the Chi-
nese Academy of Sciences, Beijing (PE) .
Flower buds used for chromosome study were fixed by
Carnoy′s solution Ⅰ ( absolute ethanol : acetic acid = 3∶1) in
middle of May, 2002 . After fixation, the flower buds were
transferred into 70% alcohol for long-term preservation . The
chromosome slides were prepared using conventional squashing
methods and stained by modified Carbol fuchsin . The micro-
graphswere taken under Leitz Orthoplan microscope using 100x
oil lens . At least five cellswerecalculated fromeach individual,
and the results were shown in Table 1 and 2 . The classification
of karyotypes followed Stebbins ( 1971 ) . Additional parameters
measuring karyotype asymmetry, i . e . , Coefficient of Karyo-
typic Overall Asymmetry (CKOA) and longest chromosome?short-
est chromosome ratio (L1?Ln ) werecalculated accordingto Zhang
(1998) .
Table 1 The parameters of chromosomes in Paeonia anomala
No <. Population Chrosome number Absolute ?. length (μm) Relative length Arm ratio Chr. Type*
1 ?13 .35±2 .09 14 .61 + 10 .96 = 25 .57 1 . 367±0 . 237 m
I 2 ?11 .00±2 .05 12 .36 + 8 .71 = 21 .07 1 . 370±0 . 267 m
Population 1 J: XJ021 3 ?9 .90±1 .72 10 .39 + 8 .58 = 18 .97 1 . 228±0 . 263 m
II 4 ?9 .68±1 .29 12 .66 + 5 .87 = 18 .53 2 . 158±0 . 224 sm
III 5 ?8 .28±0 .85 12 .17 + 3 .69 = 15 .86 3 . 300±0 . 307 st
1 ?12 .22±1 .21 13 .63 + 11 .05 = 24 .68 1 . 244±0 . 169 m
I 2 ?10 .39±0 .76 11 .99 + 9 .05 = 21 .04 1 . 346±0 . 239 m
Population 1 J: XJ027 3 ?9 .25±0 .58 10 .40 + 8 .36 = 18 .76 1 . 241±0 . 137 m
II 4 ?9 .14±0 .41 12 .29 + 6 .26 = 18 .55 1 . 959±0 . 191 sm
III 5 ?8 .36±0 .44 13 .03 + 3 .94 = 16 .97 3 . 374±0 . 640 st
1 ?11 .73±1 .67 13 .51 + 11 .21 = 24 .72 1 . 219±0 . 193 m
I 2 ?10 .31±1 .08 11 .74 + 10 .04 = 21 .78 1 . 178±0 . 129 m
Population 2 J: XJ034 3 ?9 .20±1 .44 10 .69 + 8 .69 = 19 .38 1 . 242±0 . 158 m
II 4 ?8 .41±1 .05 11 .32 + 6 .40 = 17 .72 1 . 821±0 . 330 sm
III 5 ?7 .79±1 .32 11 .85 + 4 .55 = 16 .40 3 . 088±0 . 303 st
* Chr . Type: Chromosome type, classified according Stebbins (1971)
9845 期 PAN Jin et al : Karyotypic Study on Paeonia anomala (Paeoniaceae)
Table 2 The karyotypic characters of the species reported in three sections of Paeonia
Taxon L1 ??Ln Karyotype formula Type CKOA RF
Sect K. Moutan: Subsect ?. Vaginitae
P ?. ostii T . Hong & J . X . Zhang 1 .4 2n= 2x = 10 = 6m( 3SAT) + 2sm(1SAT ) + 2st( 2SAT ) 2A 0 .248 Zhang and Hou, 1996
* P 3. jishanensis T .Hong & W .Z .Zhao 1 ?. 4 2n= 2x = 10 = 6m( 2SAT) + 2sm(2SAT ) + 2st( 2SAT ) 2A 0 .247 Hong et al , 1988
* P 3. decomposita subsp ?. decomposita 1 ?. 4 2n= 2x = 10 = 6m+ 2sm+ 2st 2A 0 .233 Hong et al , 1988
P ?. suffruticosa 1 .4 2n= 2x = 10 = 6m( 1SAT) + 2sm(1SAT ) + 2st( 2SAT ) 2A 0 .221 Hong et al , 1988
Sect K. Onaepia
P ?. californica 1 .3 2n= 2x = 10 = 6m+ 2sm+ 2st(2SAT ) 2A 0 .250 Stebbins and Ellerton,1939
P ?. brownii 1 .8 2n= 2x = 10 = 6m+ 2sm+ 2st(2SAT ) 2A 0 .273 Stebbins and Ellerton,1939
Sect K. Paeonia: 1 . subsect ,. Paeonia
P ?. anomala ( XJ021) 1 .6 2n= 2x = 10 = 6m+ 2sm+ 2st 2A 0 .245 In the text
P ?. anomala ( XJ027) 1 .5 2n= 2x = 10 = 6m+ 2sm+ 2st 2A 0 .230 In the text
P ?. anomala ( XJ034) 1 .5 2n= 2x = 10 = 6m+ 2sm+ 2st 2A 0 .198 In the text
P ?. lactiflora 1 .5 2n= 2x = 10 = 6m( 2SAT) + 2sm(1SAT ) + 2st( 1SAT ) 2A 0 .214 Hong et al , 1988
P ?. veitchii ( PB85091 - 1) 1 .6 2n= 2x = 10 = 6m( 4SAT) + 2sm(1SAT ) + 2st( 2SAT ) 2A 0 .224 Hong et al , 1988
P ?. veitchii ( PB85065 ) 1 .5 2n= 2x = 10 = 6m( 1SAT) + 2sm(1SAT ) + 2st( 1SAT ) 2A 0 .274 Hong et al , 1988
P ?. clusii ssp K. clusii ( P . 1) 1 .5 2n= 2x = 10 = 6m( 1SAT) + 2sm(1SAT ) + 2st( 1SAT ) 2A 0 .232 Tzanoudakis, 1983 (
P ?. clusii ssp K. clusii ( P . 4) 1 .5 2n= 4x = 20 = 12m+ 4sm+ 4st( 1SAT ) 2A 0 .230 Tzanoudakis, 1983 (
P ?. peregrina ( P . 27) 1 .9 2n= 4x = 20 = 12m(1SAT ) + 4sm(1SAT ) + 4st(2SAT ) 2A 0 .232 Tzanoudakis, 1983 (
2 ?. Subsect 0. Foliolatae
P ?. mairei ( PB85023 - 2) 1 .6 2n= 4x = 20 = 12m+ 4sm+ 4st( 4SAT ) 2A 0 .223 Hong et al , 1988
P ?. obovata ( PB85078) 1 .6 2n= 2x = 10 = 6m( 2SAT) + 2sm(2SAT ) + 2st( 2SAT ) 2A 0 .263 Hong et al , 1988
P ?. obovata ( PB85068) 1 .9 2n= 4x = 20 = 12m+ 4sm( 1SAT) + 4st(3SAT ) 2A 0 .217 Hong et al , 1988
P ?. obovata ( PB8524 - 4) 2 .8 2n= 4x = 20 = 12m+ 4sm+ 4st( 4SAT ) 2B 0 .175 Hong et al , 1988
P ?. mascula ssp. mascula ( P 2. 15) 1 .5 2n= 4x = 20 = 12m+ 4sm+ 4st 2A 0 .234 Tzanoudakis, 1983 (
P ?. mascula ssp. russoi ( P. 18) 1 .5 2n= 2x = 10 = 6m( 2SAT) + 2sm(1SAT ) + 2st( 2SAT ) 2A 0 .229 Tzanoudakis, 1983 (
L1 = the longest chromosome; Ln = the shortest chromosome; CKOA : Coefficient of Karyotypic Overall Asymmetry; RF: Reference
* : P . decomposita subsp. decomposita, in theoriginal referenceas P . szechuanica; P . jishanensis, in theoriginal reference as P . suffruticosa var. papaveracea .
Results
The two populations studied are diploid, 2n= 10
(Fig . 1) . The materials used for karyotypic analyses
were XJ021 ( Population 1 ) , XJ027 ( Population 1 )
and XJ034 ( Population 2 ) , representing two popula-
tions . The karyotype formulas are shown in Table 1
and 2 . The difference between these two popu-
lations is found on chromosome 3 and chromosome 4
(Table 1 ) .
The CKOA values were different in the two popu-
lations . In the two individuals of Population 1 , the
CKOA was0 .245 and 0 .230 , respectively, whilethat
of Population 2 was 0 .198 . Generally, the karyotype
of P . anamala was similar to thespecies previously re-
ported in the genus, all belonging to 2A type . The
chromosomes can be classified into three types: Type
I : including chromosome 1 through 3 , was metacen-
tric ( m) . Type II : including only chromosome 4 ,
was submetacentric ( sm) , and Type III : including
only chromosome 5 , was subtelocentric (st) .
Discussion
This was the first report of the karyotype of the
Altai populations of P . anomala . Because of the pre-
vious taxonomic confusion of this species, especially
with P . intermedia, the study addedvaluable informa-
tion to the completion of cytological data of the genus
Paeonia .
We synthesized the karyotypic parameters of all
reported species (Table 2) , and found that the karyo-
types were highly consistent in all the speciesof thege-
nus . In addition, the arm ratio of Chromosome 1 of
Sect . Moutan (1 . 43 - 1 .60) is similar in all the spe-
cies but does not overlap with that of Sect . Paeonia
(1 . 12 - 1 .37 ) , agreeing with Hong et al ( 1988 ) .
Meanwhile, the arm ratio of Chromosome 1 in Sect .
Onaepia ( 1 . 13 - 1 .20) does not overlap with that of
Sect . Moutan either . This result further confirms that
thewoody section (Sect . Moutan) has apparently dif-
ferentiated fromtheherbaceous sections (Sects . Paeo-
nia and Onaepia) with respect to the armratio of these
094 云 南 植 物 研 究 28 卷
chromosomes . The differentiation in karyotypes of the
genus also exists in the ratioof thelongest chromosome?
shortest one . In Sect . Moutan, the L1?Ln value is
1 .4 , in Sect . Paeonia L1?Ln ≥1 .5 , while in Sect .
Onaepia, L1?Ln of P . californica is 1 .3 and P . br-
ownii is 1 .8 .
图 1 P . anomala减数分裂后期 I I 图。1 , 4 : XJ021; 2 , 5 : XJ027; 3 , 6 : XJ034 .
4 , 5 , 6 分别是 1 , 2 , 3 对应的染色体组型图。标尺 = 5μm
Fig . 1 Meiotic anaphase II plates of P . anomala . 1 , 4 : XJ 021; 2 , 5 : XJ027; 3 , 6 : XJ034 .
4 , 5 , 6 is Karyogram of 1 , 2 , 3 , respectively . Bar = 5μm
In all the three sections of the genus, karyotypic
asymmetry varies slightly, showing similar CKOA val-
ues ( Table 2 ) . However, from the meiotic studies
(Sax, 1937; Stebbins and Ellerton, 1939; Walters,
1942; Zhang et al , 1997) , a great deal of abnormal
meiotic configurations was found in all the individuals
observed, indicating that there existed many chromo-
somal structural variations in the genus . These phe-
nomena are contradictory to the karyotypic consistence
recognized in Paeonia . A better understanding of the
contradiction requires further investigation . The study
raises avery interestingquestion regardingto the evolu-
tionary history of the Altai populations of P . anomala .
The diploid status of the populations does not support
the previous assumption of the allotetraploid origin of
the species . The rather conclusive molecular phyloge-
netic evidencegathered so far, however, does point to
a hybrid origin of the species ( Sang et al , 1997;
Sang and Zhang, 1999 ) . The alternative hypothesis,
therefore, is that P . anomala is a diploid hybrid that
has happened to maintain heterozygosity at the Adh lo-
ci . Given that only handful examples of well-docu-
mented diploid hybrid species were reported so far in
flowering plants ( Rieseberg, 1997 ) , this may repre-
sent an interesting case of homoploid hybrid speciation
in Paeonia . The hypothesis needs further to test with
phylogenetic data from more populations by nuclear
genes of P . anomala .
1945 期 PAN Jin et al : Karyotypic Study on Paeonia anomala (Paeoniaceae)
Acknowledgements: We thank Prof . TAN Dun-Yan (Xinjiang
Agricultural University) and Superintendent WANG Jian ( Altai
Institute of Forestry Science, Xinjiang) , and MAO Jian-Feng
and BAO Er-Jiang ( Altai Institute of Forestry Science, Xin-
jiang) for their assistance in our field works, andfor their advice
and help in the field works . This study was supported by NSFC
(Grant No . 39928003) .
References:
李懋 4学 , 张?方 , 1982 . 三倍体牡丹的细胞遗传学观察 [ J ] . 遗
传 , 4 ( 5) : 19—21
潘开 4玉 , 1979 . 中国植物志第 27 卷 [ M ] . 北京 : 科学出版 社 ,
37—59
Dark ASOS, 1936 . Meiosis in diploid and tetraploid Paeonia species
[ J ] . J Genet, 32 : 353—372
Dela ?y C, 1947 . Recheres sur la structuredes noyaux quiescentschez les
phanerogames [ J ] . Rev Cytol Cytophysiol Vdg, 9 : 169—222
Gaje ?wski W, 1948 . O Inwersji stanieu Paeonia ( Inversion hereozygosi-
ty in Paeonia) [ J ] . Acta Soc Bot Polon, 19 ( 1) : 25—55
Hong ?DY ( 洪德 元 ) , Zhang ZX ( 张志 宪 ) , Zhu XY ( 朱相 云 ) ,
1988 . Studies on the genus Paeonia ( 1) . Report of karyotypes of
somewild species in China [ J ] . Atca Phytotax Sin ( 植物分类学
报 ) , 26 : 33—43
Hong @DY ( 洪德元 ) , Pan KY (潘开玉 ) , Turland NJ , 2001 . Flora
of China: Paeoniaceae [ M ] . Beijing: Science Press and St .
Louis: Missouri Botanical Garden Press . Vol . 6 : 127—132
Hong ADY ( 洪德元 ) , Pan KY (潘开玉 ) , Li XY ( 李学禹 ) , 1994 .
Paeonia in Xinjiang, China [ J ] . Acta Phytotax Sin ( 植物分类
学报 ) , 32 ( 4) : 349—355
Okad /a H , Tamura M, 1979 . Karyomorphology and relationship in the
Ranunculaceae [ J ] . J J ap Bot, 54 : 65—77
Puni ?na EO, 1987 . Karyological study of species of the genus Paeonia
( Paeoniaceae ) from the Caucasus [ J ] . Botank Zhurnal , 72
(11) : 1504—1514
Ries ?eberg LH , 1997 . Hybrid origins of plant species [ J ] . Annu Rev
Eco Syst, 28 : 359—389
Sang ?T, Crawford DJ , Stuessy TF , 1997 . Chloroplast phylogeny, re-
ticulate evolution, and biogeography of Paeonia ( Paeoniaceae )
[ J ] . Amer J Bot, 84 : 1120—1136
Sang ?T, Zhang DM, 1999 . Reconstruct hybrid speciation using se-
quences of low cope nuclear genes: hybrid origins of five Paeonia
species based on Adh gene phylogenies [ J ] . Syst Bot, 24 ( 2) :
148—163
Sax ?K , 1937 . Chromosome inversions in Paeonia suffruticosa [ J ] . Cy-
tologia, Vol . Fujii Jub: 108—114
Sopo ?va M, 1971 . The cytological study of two Paeonia species from
Macedonia [ J ] . Fragm Balk Mus Macedon Sci Nat, 8 ( 16 ) :
137—142
Steb ?bins GL , 1938 . Cytogenetic studies in Peaonia I I . The cytology of
the diploid species and hybrids [ J ] . J Genet, 23 : 83—110
Steb ?bins GL , Ellerton S, 1939 . Structural hybridity in Peaonia califor-
nica and P . brownii [ J ] . J Genet, 38 : 1—36
Steb ?bins GL , 1971 . Chromosomal Evolution in Higher Plants [ M ] .
London: Edward Arnold Ltd
Ster ?n FC , 1946 . A Study of the Genus Paeonia [ M ] . London: The
Royal Horticultural Society
Tzan ?oudakis D, 1983 . Karyotypes of four wild Paeonia species from
Greece [ J ] . Nord J Bot, 3 ( 3) : 307—318
Walt ?ers JL , 1942 . Distribution of structural hybrid in Paeonia californi-
ca [ J ] . Amer J Bot, 29 : 270—275
Zhan ?gDM ( 张大明 ) , 1998 . Systematics of the tribe Ophiopogoneae
(Liliaceae s. 1 .) with special reference to karyotypes and chromo-
somal evolution . Cathaya 10 [ M ] . Beijing: International Aca-
demic Publishers
Zhan ?gSZ ( 张寿洲 ) , Pan KY ( 潘开玉 ) , Zhang DM (张大明 ) , et
al , 1997 . Observations on abnormal meiosis of pollen mother cells
in Paeonia suffruticosa subsp . spontanea [ J ] . Acta Bot Sin ( 植
物分类学报 ) , 39 (5) : 397—404
Zhan ?g ZP ( 张赞平 ) , Hou XG ( 侯小改 ) , 1996 . Karyotype analysis
of Paeonia ostii [ J ] . Hereditas (遗传快报 ) , 18 (5 ) : 3—6
294 云 南 植 物 研 究 28 卷