Genetic diversity and structure of natural Pinus tabulaeformis populations in North China using amplified fragment length polymorphism (AFLP)

Chinese pine (Pinus tabulaeformis carr.), endemic to China, is a conifer species with extensive and fragmented distribution in North China. In this study, the genetic diversity and structure of 20 natural populations of this species were investigated using amplified fragment length polymorphism (AFLP) markers. A total of 445 fragments were revealed with 8 pairs of primers, 379 (85.17%) of which were polymorphic. A moderate level of genetic diversity was detected at the species level (Shannon's information index I = 0.356, Nei's gene diversity H_E = 0.271) and at the population level (I = 0.219, H_E = 0.206). Most of genetic variation was within populations while a considerable level of genetic differentiation was detected (G_ST = 0.352, Ф_ST = 0.304). The high differentiation could be attributed to the complex and fragmented habitats, and a limited gene flow among populations (N_m = 0.572). The Mantel test indicated that there was significant correlation (r = 0.455, P < 0.001) between Nei's genetic distance and geographical distance among all the populations. The results suggested that proper countermeasures should be taken to prevent the habitat further deterioration and maintain the genetic diversity of this species.     

浙江省境内七子花天然种群遗传多样性研究

利用RAPD技术对浙江省境内的七子花9个天然种群遗传多样性和遗传分化进行研究.结果表明,12种随机引物对180棵植物进行检测,共得到164个可重复的位点.多态位点百分率在14.60%～27.44%(平均为20.73%),以括苍山种群最高,其次是四明山种群,最低是观音坪种群.Shannon指数和Nei指数均反映出七子花各种群具有较低的遗传多样性,但遗传分化明显.Shannon指数显示种群内遗传多样性只占总遗传多样性的27.28%,而种群间遗传多样性却占72.72%;Nei指数表明种群内的遗传变异较小,种群间的遗传变异较大,种群间的遗传分化系数为0.7157.七子花种群间的基因流为0.1987,遗传相似度平均为0.7306,遗传距离平均为0.3150,各种群间的遗传分化明显.根据遗传距离聚类分析,大致可以将9个七子花种群分为东部和西部两大类群.     

光皮桦6个南方天然群体的遗传多样性

为揭示中国特有珍贵用材树种光皮桦(Betula luminifera)天然群体的遗传多样性和遗传结构, 采用AFLP分子标记, 分析了采自浙江、福建、江西、广西和贵州5个省区6个天然群体的120份样品。9对引物获得了355个位点, 其中多态性位点323个。分析结果表明光皮桦天然群体具有较高的遗传多样性, 多态位点百分率(PPL)达90.99%, 各群体的PPL和Nei’s基因多样性(h_j)分别为93.20-98.60%和0.3143-0.3645; 总群体遗传多样性指数(H_t)为0.3616, 群体间遗传分化系数(F_st)为0.0650, 群体间总的基因流较高(N_m= 3.5962)。AMOVA分析表明群体间的遗传变异占总变异的11.49%, 浙江临安群体和贵州修文群体间的遗传距离最大(0.0665), 江西龙南群体和广西龙胜群体间的遗传距离最小(0.0173), 且遗传结构分析显示这两个群体的部分个体可能来自同一近祖。Mantel检测发现, 群体间的遗传距离与地理距离没有显著相关性(r = 0.423, P = 0.113), 而与两两群体所在地的均温差呈显著相关(r = 0.449, P = 0.017)。结合群体实地调查, 可以得出光皮桦天然群体的遗传多样性和遗传结构的形成不仅与其广域分布、自然杂交、种子特性以及生活史有关, 而且与群体被人为砍伐、生境片断化等因素有重要关系。基于上述结果我们提出了光皮桦天然种群的保护策略。     

Analysis of the genetic diversity of Bemisia tabaci populations in Shanxi province, China☆

Five different primer combinations were used for the analysis of 152 B biotype Bemisia tabaci (Gennadius) individuals and five Trialeurodes vaporairiorum individuals collected from 19 counties and seven host plants in Shanxi province in China, respectively. The main objective of the present study was to use AFLP markers to determine the genetic diversity of B. tabaci populations collected from Shanxi Province. The use of these primer combinations allowed the identification of 127 polymorphic bands (52.26%) from 60 to 500 bp. The average number of polymorphic bands per primer was 25.4 while the range for the five primers was 20–32. The average degree of heterozygosity was 0.251, while the range for the five primers was 0.204–0.289. The results suggested definite genetic diversity among different B. tabaci populations. Cluster analysis showed that B. tabaci populations were firstly scattered to three genetic groups according to the regions, then every genetic group was scattered to several subgroups according to the host plants, which revealed the genetic variability of B biotype B. tabaci populations has been not only among different regions, but also among different host plants in Shanxi Province.     

Analysis of the genetic diversity of Bemisia tabaci populations in Shanxi province, China

Five different primer combinations were used for the analysis of 152 B biotype Bemisia tabaci (Gennadius) individuals and five Trialeurodes vaporairiorum individuals collected from 19 counties and seven host plants in Shanxi province in China, respectively. The main objective of the present study was to use AFLP markers to determine the genetic diversity of B. tabaci populations collected from Shanxi Province. The use of these primer combinations allowed the identification of 127 polymorphic bands (52.26%) from 60 to 500 bp. The average number of polymorphic bands per primer was 25.4 while the range for the five primers was 20–32. The average degree of heterozygosity was 0.251, while the range for the five primers was 0.204–0.289. The results suggested definite genetic diversity among different B. tabaci populations. Cluster analysis showed that B. tabaci populations were firstly scattered to three genetic groups according to the regions, then every genetic group was scattered to several subgroups according to the host plants, which revealed the genetic variability of B biotype B. tabaci populations has been not only among different regions, but also among different host plants in Shanxi Province.     

高原鼠兔四个地理种群的遗传多样性与遗传分化

采用7 个多态微卫星DNA 标记分析了4 个高原鼠兔地理种群(海晏种群、西大滩东种群、西大滩西种群和昆仑山种群)的遗传多样性和遗传分化。采用多态信息含量(PIC)、期望杂合度(H_e )、观测杂合度(H_o)、基因流(N_m )和基因分化系数(F_st )对种群及物种的遗传多样性水平进行分析;计算各种群间的遗传距离(GD)、遗传相似度(GI)以及遗传距离与地理距离的相关系数,并进行UPGMA 聚类分析。研究结果显示,高原鼠兔的遗传多样性在小哺乳动物中处于较高水平, 其H_e 和H_o 的平均值分别为0. 5332 和0. 6003, 其F_st 为0. 0975,即有9. 75% 的变异存在于种群之间,即变异主要存在于种群内部。4 个种群间的杂合度与多态信息含量均无显著差异,说明它们的遗传多样性水平基本相当。海晏种群与其余3 个种群的遗传分化均达到了中度分化水平(F_st = 0.1043),而另3 个种群间几乎没有分化(F_st = 0. 0253)。Mantel test 结果表明地理距离与遗传距离间没有显著的相关性。     

野生扁蓿豆种质资源AFLP遗传多样性的分析

本研究采用AFLP标记对河北省、辽宁省、吉林省、山西省、内蒙古自治区和西藏自治区的10份野生扁蓿豆居群进行遗传多样性分析。从64对AFLP引物组合中,筛选出8对扩增条带清晰、多态性高的引物组合,8对引物共扩增出640个条带,其中多态性条带有472个,多态性位点百分率为73.8%;Nei's基因多样性指数平均为0.157,Shannon's多态性信息指数平均为0.099;居群间遗传相似系数(GS)平均值为0.827。聚类和主成分分析可将10个扁蓿豆居群聚为3大类,结果与居群的地理分布大致相符,呈一定的地域性分布规律。由此可见,AFLP分子标记研究结果能较好地揭示扁蓿豆居群间的遗传多样性。对我国各地的野生扁蓿豆资源的广泛收集、评价和基因资源的保护有重要的意义。     

山西省六个负蝗种群的遗传多样性

用等位酶电泳分析方法对短额负蝗(A tractom orpha sinensis)和奇异负蝗(A tractom orpha p ereg rina)各3个自然种群10种酶(AAT,CK,G 3PD,HEX,IDH,LDH,M DH,M E,PG I,PGM)进行检测。结果显示:两种负蝗在某些基因座上共享常见的等位基因,如A a t-1-b、A a t-2-b、G 3p d-a、Ck-1-b和Ldh-b;除个别基因座在部分种群符合H ardy-W e inberg平衡外,两种负蝗的大多数基因座的基因型频率显著偏离H ardy-W e inberg平衡。此外,奇异负蝗M e-c(0.318~0.740)、短额负蝗H ex-1-a(0.800~1.000)及Ldh-b(0.487~0.750)等位基因频率呈现出由北向南递增的趋势,表明M e和H ex、Ldh基因座上的等位基因频率与地理分布存在一定的相关关系。短额负蝗平均每个基因座的等位基因数(A)为1.9~2.3、多态基因座百分率(P)为56.3%~68.8%、平均观察杂合度(Ho)为0.072~0.096;而奇异负蝗的相应值依次为A=1.7~2.2,P=43.8~56.3%,Ho=0.070~0.107。从A、P和Ho3个参数可知,短额负蝗遗传多样性明显高于奇异负蝗。6个负蝗种群的平均观察杂合度均明显低于H ardy-W e inberg平衡预期值,表明6个负蝗种群均出现了杂合体缺乏现象。短额负蝗3种群I值为0.971~0.996,奇异负蝗3种群I值为0.982~0.995,短额负蝗与奇异负蝗I值为0.379~0.451,表明种内遗传相似度明显高于种间,从种间I值可知奇异负蝗和短额负蝗属于近缘种。根据R oger's遗传距离进行的聚类分析表明,两种负蝗可分为两支,且两种负蝗的遗传距离与地理距离均存在明显的相关趋势。两种负蝗的平均FST值都不显著偏离0值(奇异负蝗FST=0.087,p>0.05,短额负蝗FST=0.045,p>0.05),表明该两种负蝗种群间的分化不明显。     

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.     

Genetic diversity and structure among natural populations of Sindora glabra in Hainan Island,China as revealed by ISSR markers

Sindora glabra, one of the second-class national protective plants in China, has important ecological and commercial values. To understand the genetic diversity and structure of S. glabra, eleven natural populations from four areas of Hainan Island in China were analyzed by inter-simple sequence repeat (ISSR) markers. Thirteen primers were screened out and used to amplify 157 DNA samples. A total of 122 bands were obtained, among which 114 (93.4%) bands were polymorphic. Genetic parameters including average number of effective alleles (1.547), Nei's gene diversity (0.321), Shannon's information index (0.482), and gene differentiation coefficient (0.1944) revealed a high level of genetic diversity maintained in S. glabra populations. The variation within populations accounted for 85.6% of total variation based on analysis of molecular variance. Genetic distance analysis showed that 11 populations could be divided into three groups and populations from the same areas were classified as one group, suggesting that high genetic diversity of S. glabra was attributable to geographic isolation. Together, introduction of germplasm from distant natural distribution areas would be a sound strategy for germplasm resource conservation of S. glabra and selection of elite individuals from populations of far relationship for hybridization is of great importance for breeding improvement programs in future.     

Genetic diversity of Stipa grandis P.Smirn populations across the species’ range in the Inner Mongolia Plateau of China

In order to investigate the genetic diversity of dominant species under the background of climate change and grassland utilization in the Inner Mongolia Plateau of China, we sampled seven Stipa grandis populations along an increasing aridity gradient in the present study. The Nei’s gene diversity of populations (He) was estimated to be 0.15 and the percentage of polymorphic loci (PPL) was 49.28%. The genetic differentiation among populations (Φ_ST) was 0.2431. There was a significant relationship between genetic distance and geographic distance among the S. grandis populations by Mantel's test. The genetic diversity was significantly correlated with longitude and annual mean precipitation, which suggested that a combination of climatic factors affected the genetic diversity. The populations in the marginal habitat should be paid more attention because of their low genetic diversity and its significance for conservation of the whole species.     

北京市华北落叶松优树群体遗传多样性分析

利用SSR分子标记对北京市华北落叶松人工林5个群体的220棵优树进行遗传多样性和群体结构分析。20对SSR引物共检测到81个等位基因,每个位点等位基因数2~8个不等,平均4.05个。群体观测和期望杂合度平均值分别为0.429和0.440,Shannon信息指数和多态性信息含量分别为0.756和0.380。5个群体中,百花山和云蒙山遗传多样性水平最高,雾灵山遗传多样性水平最低。AMOVA分析结果显示,2.65%的遗传变异来自于群体间,剩余97.35%的遗传变异来自于群体内。遗传分化系数仅为0.023,表明北京市华北落叶松优树群体遗传分化程度很低。基于Nei's遗传距离可以将5个群体划分为3个类群,四海镇和雾灵山归为第Ⅰ类,松山归为第Ⅱ类,百花山和云蒙山归为第Ⅲ类。STRUCTURE群体结构分析结果与上述聚类分析结果大体一致。以上研究为华北落叶松人工林遗传多样性评价和优良种质资源收集、保护和利用提供理论依据。     

Genetic diversity and population structure of the red stingray,Dasyatis akajei inferred by AFLP marker

To examine population genetic diversity and variation of the red stingray Dasyatis akajei, samples from 1 freshwater region and 6 coastal localities within its distribution range were analyzed by using amplified fragment length polymorphism (AFLP) technology. A total of 207 loci were identified by 4 primer combinations from 87 individuals, 174 of which were polymorphic (84.1%). A high level of Nei's gene diversity was observed with the overall value of 0.230 ± 0.179. No significant genealogical clusters associated with sampling sites were revealed on the UPGMA tree. Both analysis of molecular variance (AMOVA, Φ_ST = 0.085, P = 0.00) and pairwise F_ST indicated significant genetic differentiation among four marine samples. However, no particular genetic differentiation was detected between marine and the limited sampling freshwater populations (AMOVA, Φ_CT = 0.056, P > 0.05). Except for the TZ vs. WZ (5.193), the gene flow estimates (N_m) demonstrated the effective immigrants were 1.918-2.976, suggesting low level dispersal between pairwise marine populations. Species-specific habits (demersal and sluggish habits) are probably responsible for the population structuring.     

承德大窝铺不同海拔与群落类型天然油松种群遗传多样性

利用ISSR技术分析了河北承德大窝铺3个不同海拔、4个不同群落类型的油松天然种群的遗传多样性与遗传分化,探讨了天然油松种群遗传多样性与主要生态因子的关系.结果表明:天然油松种群具有较高的遗传多样性,13个筛选出的随机引物共检测出180个位点,总多态位点比率分别达到73.33％和91.11％.海拔影响油松种群的遗传多样性,中海拔( 1070～1130 m)高于高海拔(1230 ～1350 m)和低海拔(910 ～970 m)(多态位点比率分别为63.33％、51.67％和59.44％);群落类型与种群的基因组结构变异之间存在一定的内在联系.遗传变异多来自种群内部(分别为90.45％和75.22％),种群分化不明显;群落类型对天然油松种群间遗传变异和基因交流(Gst为0.248,Nm为1.517)的影响高于海拔对其影响(Gst为0.095,Nm为4.738).可以认为,群落类型对大窝铺天然油松种群遗传分化有较大影响.     

Genetic diversity and population differentiation of the Chinese soft-shelled turtle (Pelodiscus sinensis) in three geographical populations

The Chinese soft-shelled turtle (Pelodiscus sinensis) is one of the most important economical chelonians in the world. To understand the genetic variations of the Chinese soft-shelled turtle in China, 62 individuals were sampled from three localities and 18 polymorphic microsatellite loci tested were used to detect genetic diversity and population structure. Results showed that the genetic diversity of the wild P. sinensis was high. Except for the Wuhu populations, the majority of microsatellite loci are not deviation from Hardy–Weinberg equilibrium in the other two populations. AMOVA analysis indicated that genetic variations occurred mainly within populations (97.4%) rather than among populations (2.6%). The gene flow estimates (Nm) among three geographic populations demonstrated that strong gene flow existed (Nm > 1, mean 6). The present study supported that different habitats, breed turtles escaped, multiple paternity and long evolutionary history may be responsible for the current genetic diversity and differentiation in the wild Chinese soft-shelled turtle.     

Genetic diversity of Ranunculus acris L. (Ranunculaceae) populations in relation to species diversity and habitat type in grassland communities

Correlates between genetic diversity at intra- and interpopulation levels and the species diversity in plant communities are rarely investigated. Such correlates may give insights into the effect of local selective forces across different communities on the genetic diversity of local plant populations. This study has employed amplified fragment length polymorphism to assess the genetic diversity within and between 10 populations of Ranunculus acris in relation to the species diversity (richness and evenness) of grassland communities of two different habitat types, 'seminatural' and 'agriculturally improved', located in central Germany. Within-population genetic diversity estimated by Nei's unbiased gene diversity (HE) was high (0.258-0.334), and was not correlated with species richness (Pearson's r = -0.17; P = 0.64) or species evenness (Pearson's r = 0.15; P = 0.68) of the plant communities. However, the genetic differentiation between R. acris populations was significantly correlated with the difference in species evenness (Mantel's r = 0.62, P = 0.02), but not with difference in species richness of plant communities (r = -0.17, P = 0.22). Moreover, we also found that populations of R. acris from the 'seminatural' habitat were genetically different (amova, P < 0.05) from those in 'agriculturally improved' habitats, suggesting that gene flow between these habitat types is limited. The results reported in this study may indicate that habitat characteristics influence the genetic diversity of plant species.     

Genetic diversity and structure of Manila clam (Ruditapes philippinarum) populations from Liaodong peninsula revealed by SSR markers

Manila clam (Ruditapes philippinarum) is an important commercially marine bivalve, and its wild populations have been severely declining in the coast of China during the last decade. In this study, a set of 7 genomic simple sequence repeat (SSR), and 5 expressed sequences tag (EST)-derived SSR markers were analyzed on eight wild R. philippinarum populations to assess the genetic diversity and population differentiation. A total of 114 alleles were detected on 12 loci, and the number of alleles per locus in each population ranged from 2 to 11, and allelic richness per locus varied from 2.00 to 10.88 for each sample. The average of observed and expected heterozygosity ranged from 0.386 to 0.550, and from 0.533 to 0.707, respectively. Pairwise F_ST values indicated that all population pairs had significant genetic differentiation (overall F_ST = 0.242, P < 0.01). Cluster analysis using unweighted pair group method with the arithmetic mean (UPGMA) separated the eight populations into two groups. This study will shed light on the domestication and cultivation on population genetic diversity of R. philippinarum, and also provide the foundation for conservation of R. philippinarum germplasm resources in clam breeding practices.     

竹叶眼子菜居群遗传多样性和克隆结构

采用ISSR技术对长江中游南岸豹澥湖和大冶湖不同生境中的竹叶眼子菜(Potamogeton malaianus)居群的遗传多样性及克隆结构进行了研究.结果表明,在两居群的106株个体中,利用6条ISSR引物共得到40条符合3/N标准并无连锁不平衡的清晰位点,竹叶眼子菜具有较高的遗传多样性,其多态位点百分率为75.0％,Shannon多样性指数为0.3736, 两居群的遗传分化很小.竹叶眼子菜的克隆多样性很高(D=0.9917),两居群间的克隆分化很大,不具有共有的基因型.竹叶眼子菜的基株分布为游击型构型,位于湖心的大冶居群克隆距离(3.0～31.5 m)明显地大于湖岸豹澥居群(2.4～6.7 m).     

中国二化螟不同地理种群的AFLP遗传分析

利用AFLP标记对中国11省13个二化螟Chilo suppressalis(Walker)地理种群进行遗传多态性分析,以揭示不同区域种群间遗传分化,为建立较准确的二化螟发生区划及区域治理对策提供新的依据。选取3对引物,共扩增出445个位点,其中多态性位点数为386,多态性比率为86.8%。种群间分化系数(Gst)为0.74,基因流仅为0.176;13个地理种群间遗传一致度较高,均大于0.67。遗传距离聚类分析表明,二化螟13个地理种群可区分为4类,其中淮北平原类(赣榆、阜宁、阜南)、东南山区类(鄞县、闽侯)、荆湘川类(荆州、邵阳、德阳)三类种群分别具有较一致的地理气候特征或相对较近的距离。研究还表明,与其他测试种群的遗传距离普遍较大、相对孤立的地理种群,其聚类结果有一定的随机性,因此,取样点较多、且样点分布较均匀的聚类分析结果才有较高可信度。     

Genetic diversity and population structure of the ark shell Scapharca broughtonii along the coast of China based on microsatellites

As a commercially important species in East Asia, the natural resources of Scapharca broughtonii have been suffering from severe population decline across its main habitats. In China, recovery efforts for S. broughtonii are in progress. To provide scientific bases for fisheries management and conservation program, genetic diversity and population structure of seven wild populations of S. broughtonii from the northern China coast was assessed using seven microsatellite loci in this study. High genetic diversity was present in all the seven populations, as observed in mean allelic richness per locus (11.3–12.5), and average expected heterozygosity (0.835–0.867). No significant difference in allelic richness or expected heterozygosity was observed among the seven populations. Pairwise F_ST estimates and NJ tree topologies based on D_C distances indicated that the seven populations fell into two groups, showing a clear division between the populations from the south and north of the Shandong Peninsula. Genetic differentiation was further analyzed using AMOVA and assignation tests. Genetic barrier analysis using Monmonier algorithm also confirmed that the Shandong Peninsula was the putative barrier separating the northern and southern populations. In addition, marine currents probably play an important role in high gene flow among three populations from the same marine gyre.