Genetic diversity of six populations of Atractomorpha sinensis and Atractomorpha peregrina in Shanxi Province, China

Ten enzymes (AAT,CK,G3PDH,HEX,IDH,LDH,MDH,ME,PGI,PGM)were examined using horizontal starch gel electrophoresis to estimate the levels of genetic variation within and among six natural populations of two grasshopper species Atractomorpha sinensis and A.peregrina from Shanxi,China.The collecting sites were geographically distant from each other from south to north:Quwo district,Linfen city;Xiangyuan county,Changzhi;Jinyuan district,Taiyuan city;Yuanping county,Xinzhou city and Fanshi county of Xinzhou.A.sinensis showed 43 alleles at 16 loci but A.peregrine showed 39 alleles at 15 loci (ldh-1 was deficient).The zymograms showed that some common alleles were shared at several loci in these two species (Aat-1-b,Aat-2-b,G3pdh-a,Ck-1-b and Ldh-b).However,Hex-1-a,Hex-2-a,Hex-3-a,Idh-2-b,Mdh-2-b,Mdh-1-f Pgi-b,Pgm-b had common alleles in A.sinensis and Hex-1-b,Hex-2-b,Hex-3-b,Idh-2-a,Mdh-2-a,Mdh-1-d,Pgi-a,Pgm-c were of high frequency in A.peregrine instead.Most of the observed genotype frequencies were found to significantly deviate from the Hardy-Weinberg expectations in both species.A tendency of clinal distribution of allele frequency was observed at three loci.The frequency of the moderately migrating allele Me-c (0.318-0.740)in A.peregrina,Hex-1-a (0.800-1.000)and Ldh-b (0.487-0.750)in A.sinensis demonstrated increased frequency from north to south.Such tendency suggests that the allele frequency in these three loci may be correlated with the species'geographic distributions.A.sinensis showed higher genetic diversity than A.peregrina as indicated by higher mean number of alleles per locus (A=1.9-2.3 in A.sinensis and 1.7-2.2 in A.peregrina),percentage of polymorphic loci (56.3%-68.8%in A.sinensis and 43.8%-56.3%in A.peregrina),and the observed heterozygosities (Ho=0.072-0.096 in A.sinensis and 0.070-0.107 in A.peregrina).The observed heterozygosities of the six populations were all noticeably lower than the Hardy-Weinberg expectations,mostly due to heterozygote deficiency in the populations of both species.The overall mean Fsr were small (FST=0.045,P＞0.05 in A.sinensis populations and 0.087,P＞0.05 in A.peregrina populations).Nei's genetic identity (I)estimates indicate low intraspecific (＞0.95)but higher interspecific (0.377-0.447)genetic diversity.The cluster analysis based on modified Roger's genetic distance (D)showed that the two species were divided into two branches.Both species are of limited dispersal capacity and a moderate geographical barrier might significantly restrict the gene exchange among populations,resulting in accumulation of local genetic differentiations.The A.sinensis populations used in this study were separated from each other by 155.2 to 271.4 km and the A.peregrina populations were separated from each other by 78.8 to 174.9 km with observable physical barriers.The aUozyme data showed only minimal genetic differentiation at population level,most likely as a result of gene exchange.It is reasoned that natural factors and human agricultural activities might have facilitated migration and dispersal for the two species.     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D. dyeriana. Based on random amplified polymorphic DNA (RAPD) markers, a comparative study of the genetic diversity and genetic structure of Dipteronia was performed. In total, 128 and 103 loci were detected in 17 D. sinensis populations and 4 D. dyeriana populations, respectively, using 18 random primers. These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%, respectively, indicating that the genetic diversity of D. sinensis was higher than that of D. dyeriana. Analysis, based on similarity coefficients, Shannon diversity index and Nei gene diversity index, also confirmed this result. AMOVA analysis demonstrated that the genetic variation of D. sinensis within and among populations accounted for 56.89% and 43.11% of the total variation, respectively, and that of D. dyeriana was 57.86% and 42.14%, respectively. The Shannon diversity index and Nei gene diversity index showed similar results. The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high. Analysis of the genetic distance among populations also supported this conclusion. Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon. The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D. sinensis (p < 0.01), while no significant correlation was found between genetic and geographical distances among populations of D. dyeriana. This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale. This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges. We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats. Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination, and to help conserve genetic diversity. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(5): 785–792 [译自: 植物生态学报, 2005, 29(5): 785–792]     

Genetics and polymorphism of a duplicated malate dehydrogenase (MDH) locus in the grasshopperOxya japonica japonica (Orthoptera:Acrididae:Oxyinae)

A biochemical genetic study of the enzyme malate dehydrogenase (MDH) was conducted in the grasshopperOxya j. japonica. Analysis of MDH electrophoretic variation in this species of grasshopper shows that one of the two autosomal loci for MDH in grasshoppers, the Mdh-2 locus, controlling the anodal set of MDH isozymes, is duplicated. Results of breeding studies confirm this and the observed polymorphism at theMdh-2 locus in the two populations ofOxya j. japonica studied can be attributed to three forms of linked alleles at the duplicated locus in equilibrium in both populations. In this respect, all individuals of this species possess heterozygous allelic combinations at the duplicatedMdh-2 locus, which may account for the spread of the duplicated locus in the populations of this species of grasshopper.This research was supported by a grant (Vote F) from the University of Malaya, Kuala Lumpur.     

Isolation and characterization of 11 microsatellite loci from Camellia sinensis in Taiwan using PCR-based isolation of microsatellite arrays (PIMA)

We report 11 novel microsatellite primer pairs for the wild tea, Camellia sinensis (L.) O. Kuntze forma formosensis Kitamura. These simple sequence repeat markers were tested in 24 samples collected from wild tea populations, and in cultivars and C. japonica. The number of alleles ranged from 4 to18. The expected (H _E) and observed (H _O) heterozygosity were 0.687–0.946 and 0.042–0.792, respectively. All loci were significantly deviated from Hardy-Weinberg expectations due to the heterozygote deficiency, indicating a dramatic loss of genetic polymorphisms in the rare species. Significant LD was discovered in most loci. These primers may provide a tool for understanding demography and population structure in wild tea.     

云南松居群遗传学研究的等位酶分析方法

针对１５个云南松Ｐｉｎｕｓｙｕｎｎａｎｅｎｓｉｓ居群,开展了１４种酶系统的水平切片淀粉凝胶电泳实验,在谱带遗传分析的基础上确定了３３个等位酶位点及其等位基因。其中有３２个等位酶位点是多态的（有２个以上的等位基因）,只有一个单态位点Ｄｉａ－４。有３个等位基因的位点有Ｌａｐ－１、Ｌａｐ－２、Ａａ－３、Ｓｋｄ－１、Ｓｋｄ－２、Ａｄｈ－１、Ａｄｈ－３、Ｇｄｈ、Ｐｇｄ－１、Ｐｇｍ－１、Ｐｇｍ－３、Ｐｇｉ－１、Ｐｇｉ－３、Ｍｄｈ－１、Ｍｅ、Ｇ６ｐｄ、Ｄｉａ－１、Ｔｐｉ－１、Ｔｐｉ－２、Ｔｐｉ－３和Ｔｐｉ－４,有４个等位基因的位点有Ｓｋｄ－３、Ａｄｈ－２、Ｐｇｄ－２、Ｍｄｈ－２、Ｍｄｈ－３、Ｍｄｈ－４和Ｄｉａ－２,有５个等位基因的位点有Ａａｔ－１和Ｄｉａ－３。云南松居群的等位基因平均数Ａ＝２１,在松属中居于中上水平。本研究揭示了云南松居群酶位点及其等位基因带谱的变异式样,为松属植物的遗传多样性研究提供了一批酶位点及其等位基因的参考图谱     

Development of 15 polymorphic microsatellite loci from <Emphasis Type="Italic">Garrulax morrisonianus</Emphasis> (Timaliidae), an endemic avian species of Taiwan

Garrulax morrisonianus, an endemic avian species of Taiwan, inhabits evergreen forests at high elevations of 2,000–3,952 m. In this study, we developed 15 microsatellite primer pairs for genetic study. These markers were screened for 52 samples collected from wild populations of different geographical regions. The number of alleles ranged from 4 to 13. The expected (H _E) and observed (H _O) heterozygosities were 0.430–0.725 and 0.000–0.500, respectively. All loci were significantly deviated from Hardy-Weinberg equilibrium due to the heterozygote deficiency. The authors Tzen-Yuh Chiang, Sao-Cheng Lao, Ya-Fu Lee contributed to the study equally.     

Subpopulation genetic structure of a plant panmictic population of Castanea sequinii as revealed by microsatellite markers

Castanea squinii Dode, an endemic tree widely distributed in China, plays an important role both in chestnut breeding and forest ecosystem function. The spatial genetic structure within and among populations is an important part of the evolutionary and ecological genetic dynamics of natural populations, and can provide insights into effective conservation of genetic resources. In the present study, the spatial genetic structure of a panmictic natural population of C. sequinii in the Dabie Mountain region was investigated using microsatellite markers. Nine prescreened microsatellite loci generated 29–33 alleles each, and were used for spatial autocorrelation analysis. Based on Moran’s I coefficient, a panmictic population of C. sequinii in the Dabie Mountain region was found to be lacking a spatial genetic structure. These results suggest that a high pollen-mediated gene flow among subpopulations counteract genetic drift and/or genetic differentiation and plays an important role in maintaining a random and panmictic population structure in C. sequinii populations. Further, a spatial genetic structure was detected in each subpopulation’s scale (0.228 km), with all three subpopulations showing significant fine-scale structure. The genetic variation was found to be nonrandomly distributed within 61 m in each subpopulation (Moran’s I positive values). Although Moran’s I values varied among the different subpopulations, Moran’s I in all the three subpopulations reached the expected values with an increase in distances, suggesting a generally patchy distribution in the subpopulations. The fine-scale structure seems to reflect restricted seed dispersal and microenvironment selection in C. sequinii. These results have important implications for understanding the evolutionary history and ecological process of the natural population of C. sequinii and provide baseline data for formulating a conservation strategy of Castanea species. __________ Translated from Acta Phytoecologica Sinica, 2006, 30(1): 147–156 [译自: 植物生态学报]     

基于SSR标记的大明松天然群体遗传多样性分析

大明松是广西和贵州特有的高山松类,具有很高的经济价值和生态价值,其自然种群长期没有得到充分的保护和利用,不利于该树种长期稳定发展。为了合理保护和开发利用大明松天然遗传资源,该文利用12个SSR分子标记对大明松3个天然群体进行遗传多样性研究,分析其群体间的遗传分化和基因流,为该物种的保护策略提供参考。研究结果表明:(1)12对引物共检测到37 个等位基因,多态位点百分率为100%。每个位点平均观察等位基因数(Na)为3.08,平均有效等位基因数(Ne)为1.68, 不同位点间有效等位基因数差异较大; 每个位点平均观察杂合度(Ho)为0.35,平均期望杂合度(He)为0.40,平均多态信息含量(PIC)为0.31。(2)3 个群体的Shannon''s多样性指数变化范围为0.48～0.65,Nei''s基因多样度的变化范围为0.27～0.39,与其他近缘种松类相比遗传多样性偏低。群体平均观察杂合度为0.40,平均期望杂合度为0.33,平均有效等位基因数为1.58。群体间基因分化系数(Gst)为0.10,群体间的遗传分化水平较低,大部分变异均存在群体内。群体间的基因流(Nm)为2.74,说明大明松群体间的基因交流比较充分。该研究可为大明松生物多样性保护提供重要参考依据,为科学利用大明松资源打下基础。     

Polymorphic microsatellites identified by cross-species amplifications in the European Coot Fulica atra

We tested the cross-amplification of 26 microsatellites developed for passerines and an additional three developed for Gallinula species in eight European Coots from two populations. Sixteen microsatellite markers successfully amplified, of which nine were polymorphic with 2–6 alleles (mean 3.7 alleles) and an expected heterozygosity (H _e) ranging from 0.375 to 0.805 (mean H _e = 0.589). On average, we found 2.22 alleles/locus and a mean H _e of 0.440 in one nest, and 2.56 alleles/locus and a mean H _e of 0.494 in the other one. These nine polymorphic markers could be of potential use in studies of genetic variability, population structure and reproductive strategy of European Coots.     

Isozyme variation in Larrea ameghinoi and Larrea nitida (Zygophyllaceae): genetic diversity and its bearing on their relationship

Electrophoretic variants for 11 isozyme systems were examined by horizontal polyacrilamide gel electrophoresis in natural populations of Larrea ameghinoi and L. nitida (section Larrea). The two taxa presented fixed alternative allelic variants at loci Adh-1, Gdh-1 and Mdh-1. Genetic variability estimates showed no statistically significant differences among populations of section Larrea (H_e: 0.097–0.167). However, these values were significantly lower than those reported for species of section Bifolium (L. divaricata, diploid L. tridentata) in a previous contribution (H_e: 0.17–0.29). Positive and highly significant Wright's fixation indices are in agreement with higher amounts of inbreeding for nitida and L. ameghinoi, as suggested on the basis of floral morphology by previous authors. Moreover, the higher levels of genetic differentiation obtained for conspecific populations of section Larrea as compared to those of section Bifolium are also consistent with these observations. Nei's genetic identity values obtained for sympatric (I=0.80) and allopatric (I=0.63–0.73) L. ameghinoi – L. nitida population pairs are concordant with those expected for different, but closely related congeneric species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Isolation and characterization of 12 microsatellite loci from <Emphasis Type="Italic">Suzukia shikikunensis</Emphasis> (Lamiaceae), a genus endemic to Taiwan and Ryukyus

Suzukia shikikunensis Kudo is an endemic plant in Taiwan and suffers habitat destruction caused by human overexploitation. In this study, we developed 12 microsatellite primer pairs for genetic study. These markers were screened for 24 samples collected from wild populations distributed in Taiwan, and for a sister species S. luchuensis collected from Yonaguni and Lutao Islands. The number of alleles ranged from 5 to 14. The expected (H _E) and observed (H _O) heterozygosities were 0.65–0.922 and 0–0.625, respectively. All loci were significantly deviated from Hardy–Weinberg equilibrium due to the heterozygote deficiency. These primers amplifying microsatellites in the two species may provide a useful tool for population genetics to establish conservation strategy.     

Utility of RAPD markers in identifying genetic linkages to genes of economic interest in peach

The identification of molecular markers linked to economically important traits for use in crop improvement is very important in long-lived perennial species. Three-hundred-and-sixty RAPD primers were used with bulked segregant analysis to identify markers linked to loci of specific interest in peach [(Prunus persica) L. Batch] and peach x almond [(Prunus dulcis) Batch] crosses. The traits analyzed included flesh color, adhesion, and texture; pollen fertility; plant stature; and three isozyme loci. The Mendelian behavior of the RAPD loci was established, and RAPD markers were mapped relative to the loci controlling flesh color, adhesion, and texture, and the isozyme loci Mdh-1, 6Pgd-2 and Aat-1, as well as the existing RFLP genetic linkage map constructed previously using a peach x almond F₂ population. This technique has facilitated rapid identification of RAPD and RFLP markers that are linked to the traits under study. Loci controlling these traits mapped predominantly to linkage groups 2 and 3 of the peach genetic linkage map. Linkages to genes with both dominant and co-dominant alleles were identified, but linkages to dominant genes were more difficult to find. In several crosses, RAPD marker bands proved to be allelic. One co-dominant RAPD formed a heteroduplex band in heterozygous individuals and in mixtures of alternate homozygotes. The Mendelian behavior of the RAPD loci studied was established and the results suggest that RAPD markers will be useful for plant improvement in peach.     

Fruits foraging patterns and seed dispersal effect of frugivorous birds on Hippophae rhamnoides sinensis

Behaviors of 18 species of birds eating fruits of Hippophae rhamnoides spp. sinensis were observed from September 2003 to March 2004. Their foraging patterns were found to be very different and can be divided into five classes: (1) direct swallowing the fruits on crown of the shrubs and sometimes regurgitating seeds soon after; (2) carrying the fruits to their perching sites and swallowing; (3) pecking the fruits from the shrubs to the ground, eating pulp and seeds but leaving pericarp; (4) pecking through the pericarp, eating pulp and leaving pericarp and seeds; (5) pecking through the pericarp on the top of fruits, and only eating seeds. These foraging patterns have different effects on seed dispersal of H. rhamnoides spp. sinensis. The germination experiment of three groups of seeds (seeds from feces, dry fruits and extracted seeds from dry fruits) was carried out. Although ingestion processes of birds had some adverse effects on the seed germination of H. rhamnoides spp. sinensis, the seeds from feces still have a relatively higher germination ratio. H. rhamnoides spp. sinensis provides food to a variety of frugivorous birds, and the birds disperse its seeds. Thus, a mutually beneficial relationship between the bird and the seed is formed. __________ Translated from Chinese Journal of Ecology, 2005, 24(6): 635–638 [译自: 生态学杂志, 2005, 24(6): 635–638]     

Isolation and characterisation of microsatellite loci in Calystegia soldanella (Convolvulaceae), an endangered coastal plant isolated in Lake Biwa,Japan

Eight microsatellite loci of Calystegia soldanella useful for comparisons of the genetic structure of isolated populations in the ancient Lake Biwa and coastal populations in Japan were isolated and characterised. The number of alleles ranged from 2 to 5. The expected (H _E) and observed (H _O) heterozygosities were 0.097–0.583 and 0.000–0.380, respectively, from 100 individuals from Lake Biwa and coastal populations. Seven of the eight loci exhibited significantly fewer heterozygotes than expected based on the Hardy–Weinberg equilibrium (P < 0.05). These primers amplifying microsatellites in C. soldanella may provide a population genetics tool useful in the establishment of a conservation strategy.     

贵州省野生皂荚种实表型性状多样性分析

为深入了解贵州省野生皂荚(Gleditsia sinensis)荚果表型性状的遗传多样性及其变异类型,为皂荚的遗传改良、种质鉴定、亲本选择以及品种培育奠定理论基础。该研究以贵州省7个野生皂荚群体70个个体为研究对象,采用方差分析、主成分分析、相关性分析及多性状综合评价等方法对皂荚群体的10个种实表型性状进行系统分析和综合评价。结果显示：(1)所测皂荚的表型性状差异在群体内均达到极显著水平(P＜0.01);在群体间,除每荚粒数、种子宽、种子长宽积以及种子长宽比之外,其余表型性状的差异均达极显著水平(P＜0.01)。(2)7个居群野生皂荚各性状平均变异系数为21.16%,其中凯里市(P4)居群的变异系数最高(24.44%);居群间荚果的变异(29.22%)高于种子的变异(11.04%),且变异主要来自于群体内。(3)相关分析显示,皂荚种实各性状之间存在不同程度的关联性;主成分分析显示,前4个主成分(皂荚种子大小、单个荚果出籽数量、种子形态指数因子、与荚果长和种子厚相关的因子)的累积贡献率达69.783%,可基本反映皂荚表型性状的大部分信息;以10个种实性状对皂荚野生群体进行综合评价发现,来自于惠水县(P7)群体的皂荚种实性状综合评价最高。研究表明,贵州省野生皂荚在群体间及群体内具有丰富的表型变异,且群体内的变异大于群体间的变异,变异主要来自于群体内。     

Assessment of genetic diversity and population structure of Chinese wild almond, Amygdalus nana, using EST- and genomic SSRs

Amygdalus nana L., commonly known as wild almond, is an endangered wild relative of cultivated almond, which has great potential in almond crop breeding. In this study, we used microsatellite (SSR) loci derived from both expressed sequence tag (EST) and anonymous genomic sequence to explore the genetic diversity and population structure of A. nana in Xinjiang of China. Seven natural populations were collected across the whole distribution of A. nana in China, including populations from both inside (four populations) and outside (three populations) the established protected areas. A total of 22 and 19 alleles were detected from the seven pairs of EST and genomic SSR loci, respectively. Generally, the genomic SSRs showed lower levels of variation than EST-SSRs, which may partially due to the higher cross-species transferability in EST-SSRs than in genomic SSRs. The population-level genetic diversity (A = 1.84, P = 50.00%, H_o = 0.3491, H_E = 0.2271) was lower than cultivated almond and several wild fruit species with similar breeding system. Most of the genetic variation (82.16%) was partitioned within populations. In particular, the population collected from Tacheng County (outside the protected areas) had the highest levels of genetic diversity and had significantly different genetic constitution from other populations.     

Isolation of microsatellite markers in the Emys orbicularis complex and development of multiplex PCR amplification

We report the development of 15 new microsatellite markers for Emys orbicularis and Emys trinacris. A survey of 20 individuals showed that all loci are highly polymorphic with 3–14 alleles per locus. Additionally, 22 Glyptemys muhlenbergii primers were checked for cross-species amplification, with 14 being amplified successfully and polymorphic (2–14 alleles). A set of eight markers was selected and combined into two multiplex PCRs. Levels of genetic diversity were assessed in 648 individuals covering the complete distribution area. The number of alleles ranged from 13 to 24 and observed heterozygosity varied between 0.515 and 0.852.     

Isolation and characterization of polymorphic microsatellite loci from a repeat-enriched genomic library of stone flounder (Kareius bicoloratus) and cross-species amplification

Stone flounder (Kareius bicoloratus) is a rare fish species in China. Here, we reported 11 polymorphic microsatellite loci isolated from a repeat-enriched genomic library of stone flounder. The number of alleles, observed and expected heterozygosity per locus in 32 individuals ranged from 3 to 6, from 0.1613 to 0.8667 and from 0.1549 to 0.7932, respectively. Two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium between pairs of loci was found. Cross-species amplification of these microsatellite loci in additional three fish species was performed. These polymorphic microsatellite loci would be useful for investigating genetic population structure and construction of genetic linkage map in Kareius bicoloratus. Yong-Sheng Tian and Gui-Dong Miao are contributed equally.     

Development and characterization of polymorphic microsatellite loci in endangered fern Adiantum reniforme var. sinense

Fourteen polymorphic microsatellite loci in the endangered fern Adiantum reniforme var. sinense were developed and characterized using the Fast Isolation by AFLP of Sequences Containing repeats (FIASCO) protocol. Polymorphism of each locus was assessed in a bulked sample of 30 individuals from 8 natural populations. The number of alleles per locus varied from 2 to 11, with an average value of 6.2. The ranges of observed and expected heterozygosity were 0.000–0.895 and 0.226–0.868, respectively. These microsatellite markers provide useful tools for the ongoing conservation genetic studies of Adiantum reniforme var. sinense.     

Genetic structure and differentiation of Psathyrostachys huashanica populations detected with RAPD markers

Psathyrostachys huashanica Keng is a perennial grass and belongs to genus Psathyrostachys under Triticeae. sathyrostachys is found in the center of Middle Asia and the Caucasus Mountain, while P. huashanica, a species endemic to China, is only located in Mt. Hua in the Shaanxi province, China. At present, the population of this species is decreasing, and reaching the edge of extinction. Due to the limitation in distribution and the importance as breeding material for germplasm storage, it has been considered as first class among the national protected rare plants. For this reason, the present study is significant in probing plant flora, origin and evolution of Triticeae, and crop breeding. Randomly amplified polymorphic DNA (RAPD) markers were used to analyze the genetic structure and differentiation of P. huashanica populations sampled in three valleys (Huangpu, Xian, and Huashan Valleys) in Mt. Hua. One hundred and twenty-two RAPD fragments were obtained in all 266 individuals with 20 primers with a mean of 6.1 (2–10) fragments per primer. The percentage of polymorphic loci (PPB) was 60.66% in Huangpu Valley, 90.98% in Xian Valley, 95.08% in Huashan Valley, and the total PPB was 95.08%, which indicated a highly genetic variability of P. huashanica. The Shannon’s Information index and G _ST were 0.3306 and 0.3263, respectively, indicating that there were more genetic variations within the subpopulations than those among the subpopulations. The gene flow among the subpopulations of P. huashanica (Nm = 1.0322) was much less than that of the common anemophytes (Nm = 5.24). Mean genetic distance is 0.1 571 (range: 0.0022–0.2901). The highest value of genetic distance was found between the subpopulation (hp1) of Huangpu Valley and the highest altitude subpopulation (h8) of Huashan Valley. Correlation analysis detected significant correlation between genetic distance and vertical distance of altitude. Clustering analysis and principal coordinate analysis revealed the genetic differentiation among the populations of P. huashanica. Differentiation mainly occurred between the higher altitude subpopulations and the lower altitude subpopulations, suggesting that altitude might be the major factor that restricted the gene flow between different altitude subpopulations and resulted in differentiation of subpopulations. Translated from Acta Ecologica Sinica, 2005, 25(4): 719–726 [译自: 生态学报]