Genetic diversity analysis by RAPD in Cathaya argyrophylla Chun et Kuang

Genetic diversity level ofCathaya argyrophylla was confirmed by random amplified polymorphic DNA (RAPD) markers. Seventy five samples (individuals), collected from Hunan and Sichuan provinces of China were used in the study. 21 10-mer oligonucleotide primers detected 106 sites, and 34 (32%) of them were polymorphic. The level of genetic variation in C.argyrophylla was lower than those of other conifers, and was considered to be associated with the complexity of habitats. The percentages of polymorphic sites (PPS) in the Hunan and Sichuan populations were 18% and 25% respectively. 7.99% of genetic variation existed between the two populations; this value was higher than the mean value (6.8%) among populations in conifers displayed by allozyme. Some subpopulations ofC. argyrophylla were greatly differentiated because of site mutation and genetic drift. The highest value of genetic difference between subpopulations amounted to 16. 23%. In addition, a concept of diversity coefficient (DC), a value used to measure the genetic diversity level, and its calculation were proposed. The low genetic diversity level ofC. argyrophylla was thought to be one of the factors causing its endangered status. Project supported by the National Natural Science Foundation of China.     

Genetic diversity analysis by RAPD in Cathaya argyrophylla Chun et Kuang

Genetic diversity level of Cathaya argyrophylla was confirmed by random amplified polymorphic DNA (RAPD) markers. Seventy five samples (individuals), collected from Hunan and Sichuan provinces of China were used in the study. 21 10-mer oligonucleotide primers detected 106 sites, and 34 (32% ) of them were polymor-phic. The level of genetic variation in C. argyrophylla was lower than those of other conifers, and was considered to be associated with the complexity of habitats. The percentages of polymorphic sites (PPS) in the Hunan and Sichuan pop-ulations were 18% and 25% respectively. 7.99% of genetic variation existed between the two populations; this value was higher than the mean value (6.8%) among populations in conifers displayed by allozyme. Some subpopulations of C. argyrophylla were greatly differentiated because of site mutation and genetic drift. The highest value of genetic dif-ference between subpopulations amounted to 16. 23% . In addition, a concept of diversity coefficient(DC), a value us     

Genetic variation within the endangered quillwort Isoëtes hypsophila (Isoetaceae) in China as evidenced by ISSR analysis

The genetic diversity of 56 individuals of Isoëtes hypsophila Hand.-Mazz. from China was investigated by ISSR. Twelve primers were screened from 65 primers, and a total of 119 DNA fragments were scored, of these, 82% were polymorphic bands, which indicated that high levels of genetic variation existed in the natural populations. Genetic diversity varied greatly among populations with the percentage of polymorphic band (PPB) values ranging from 8 to 35%. An analysis of molecular variance (AMOVA) was used to apportion the variation between regions, among populations within regions, and within populations. Results indicated that most of the variance (85%) occurred between Yunnan and Sichuan. The variances among populations within regions and within populations, however, were only 5 and 10%, respectively. In the among-population analysis, the larger part of genetic variation (77%) resided among populations, and less (23%) presented differences within populations. UPGMA cluster analysis showed that there was no distinct genetic differentiation between populations from Sichuan province. A number of causes including limited gene flow, genetic drift and inbreeding might have led to these observed genetic profiles of I. hypsophila.     

Genetic diversity and population structure of the endangered species Psammosilene tunicoides revealed by DALP analysis

Psammosilene tunicoides W. C. Wu et C.Y. Wu (Caryophyllaceae) is a rare plant endemic to China. We used the direct amplification of length polymorphism (DALP) method to analyse the genetic diversity of 17 natural populations of this species from Yunnan, Sichuan and Guizhou provinces. The percentage of polymorphic bands (PPB) was 99.1% at the species level and 19.9% at the population level. The total genetic diversity (Ht) was 0.2827; the genetic diversity within populations (H_s) was 0.0594 and the coefficient of gene differentiation (G_st) was 0.7899. The results indicated low levels of genetic diversity within populations and extremely high genetic differentiation among populations. This pattern might be caused by genetic drift resulting from intensive harvesting and habitat fragmentation. It might also be a result of local adaptation. We further performed experiments on its breeding system and found a high rate of selfing in P. tunicoides, which may be partially responsible for its population structure.     

Genetic Diversity Within and Among Lepidium draba Populations from Eastern Anatolia Based on RAPD Analysis

Genetic variation and structure of six natural populations of Lepidium draba L. from Eastern Anatolia were assessed using random amplified polymorphic DNA (RAPD) markers. For RAPD analysis, 12 primers generated 218 reproducible bands across the six populations analyzed, of which 73 bands (33.3%) were polymorphic. The mean Nei’s gene diversity value for all six populations was 0.1771. Shannon’s information index varied with population (0.2278–0.3082), averaging 0.2608. Analysis of molecular variance (AMOVA) showed that genetic diversity was greater within populations (58.66%) than among populations (30.68%). In addition, the variation between groups was 10.33%. The genetic differentiation among populations (G _ST) was 0.3210, indicating that most genetic diversity occurs within populations. Gene flow (Nm) was low, at only 0.5288.     

ISSR analysis of genetic diversity of the Qinghai-Tibet Plateau endemic Rhodiola chrysanthemifolia (Crassulaceae)

Inter-simple sequence repeat markers (ISSR) were used to estimate genetic diversity within and among 10 populations of Rhodiola chrysanthemifolia along Nianqingtangula Mountains and Brahmaputra, a species endemic to the Qinghai-Tibet Plateau and an endangered medicinal plant. Of the 100 primers screened, 13 produced highly polymorphic DNA fragments. Using these primers, 116 discernible DNA fragments were generated of which 104 (89.7%) were polymorphic, indicating substantial genetic diversity at the species level. Genetic diversity measured by the percentage of polymorphic bands (PPB) at the population level ranged from 21.97% to 48.8%. Analysis of molecular variance (AMOVA) showed that the genetic variation was found mainly among populations (77.3%), but no regional differentiation was discernible. Variance within populations was only 22.7%. The main factor responsible for this high level of differentiation among populations is probably the historical geographical and genetic isolation of populations in a harsh mountainous environment. Concerning the management of R. chrysanthemifolia, the high genetic differentiation of populations indicates the necessity of conserving the maximum possible number of populations.     

珍稀濒危植物长叶红砂种群遗传多样性的ISSR分析

采用ISSR分子标记技术,对濒危小灌木长叶红砂(Reaumuria trigyna) 集中分布的5个种群的遗传多样性水平和遗传结构进行了研究。14条引物共检测到114个位点,其中99个为多态位点,多态位点比率为86.84％,长叶红砂种群具有较高的遗传多样性。物种水平上Shannon多样性指数(I)为0.468 8,Nei基因多样性指数(H)为0.308 4;种群水平上,多态位点比率P为77.89%,I为0.410 6,H为0.260 9,基因分化系数Gst为0.106 9,揭示了长叶红砂种群遗传变异多存在于种群内,种群间的遗传分化较小,占10.69%。 基因流(Nm)为4.178 7＞1,说明种群间的基因交流,防止了由于遗传漂变导致的遗传分化。聚类分析表明长叶红砂种群遗传距离与地理距离之间无显著的相关性。研究结果说明遗传多样性水平与物种本身特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低。     

Genetic diversity in the northernmost<Emphasis Type="Italic"> Oryza rufipogon</Emphasis> populations estimated by SSR markers

To estimate genetic diversity of the residual northern populations of Oryza rufipogon, a total of 232 individuals from six populations were analyzed using microsatellites (SSRs). The O. rufipogon populations with different status included three from Dongxiang (Jiangxi Province) and three from Chaling (Hunan Province) in China. The 23 rice SSR primer pairs selected from the RiceGenes Database detected a total of 115 alleles, indicating that all the SSR loci were polymorphic in this study. The total gene diversity was 0.919 in the six O. rufipogon populations, and the Donxiang populations showed higher diversity than the Chaling populations. More significant genetic differentiation and less gene flow were found among the Dongxiang populations than those from Chaling. The two putative introgressed populations showed relatively high genetic variation. One in situ conserved population from Dongxiang had the lowest level of genetic diversity. The re-introduced population from Chaling restored about 90% of the genetic variation, compared with the original source population. It is concluded from these results that a relatively high level of genetic variation resided in the northern O. rufipogon populations and continued efforts of conservation of these populations are needed; and that the conservation of some Chaling and Dongxiang populations has been effective in preventing gene flow from cultivated rice. Introgression of cultivated rice demonstrated significant impacts on genetic variability of the O. rufipogon populations, and should be carefully considered in conserving this wild rice. This study also suggested that re-introduction to its original habitats is an effective approach to restore O. rufipogon populations.Communicated by J.S. Heslop-Harrison     

拟环纹豹蛛不同地理种群遗传多样性的AFLP标记研究

为查明拟环纹豹蛛Pardosa pseudoannulata不同地理种群的遗传多样性机制,应用AFLP技术对6个拟环纹豹蛛地理种群的遗传多样性进行了研究分析。8对引物组合扩增出1 038个AFLP条带,其中多态性条带占86.622%,全部个体显示了各自独特的AFLP图谱。AFLP标记的遗传多样性分析结果表明： 拟环纹豹蛛无论在物种水平(P=86.62%,H=0.2622,I=0.3101),还是在种群水平（P=73.0%,H=0.2155,I=0.2554）都表现出较高的遗传多样性。其中湖南长沙雷锋镇种群内遗传变异最大,云南高黎贡山福贡种群内遗传变异最小,华南北部（湖南、湖北、江西）地区拟环纹豹蛛遗传多样性明显高于华南南部（云南、海南）种群。据种群变异来源分析,有35.77%的遗传变异来自种群间,64.23%的变异来源于种群内（N_m=0.898）,不同地理种群显示出一定的遗传分化。分析认为海拔是影响拟环纹豹蛛遗传分化的重要因素,这为进一步明确我国稻田狼蛛优势种群在农药胁迫下的遗传适应性机制提供了实验依据。     

Genetic diversity of Sinojackia dolichocarpa (Styracaceae), a species endangered and endemic to China,detected by inter-simple sequence repeat (ISSR)

Sinojackia dolichocarpa, a species endangered and endemic to China, is distributed only in the regional area of Shimen and Sangzhi in Hunan Province. Inter-simple sequence repeat (ISSR) markers were used to investigate the genetic diversity within and among the four natural populations of S. dolichocarpa. Leaf samples were collected from 84 individuals. Thirteen ISSR primers selected from 80 primers gave rise to 137 discernible DNA bands of which 100 (72.99%) were polymorphic. On average each primer gave rise to 10.5 bands including 7.7 bands with polymorphic profile. At the species level, high genetic diversity was detected (PPB: 72.99%; H_E: 0.2255; Ho: 0.3453). However, relatively low genetic diversity existed within populations. Population in Maozhuhe (MZH) exhibits the greatest level of variability (PPB: 40.38%, H_E: 0.1566, Ho: 0.2330), whereas the population in Jingguanmen (JGM) finds its own variability at the lowest level (PPB: 30.66%; H_E: 0.1078; Ho: 0.1601). A high level of genetic differentiation among populations was revealed by Nei's gene diversity statistics (45.30%), Shannon's information measure (45.24%) and analysis of molecular variance (AMOVA) (52.88%). The main factors responsible for the high level of differentiation among populations are probably related to the selfing reproductive system and the isolation of populations. The strong genetic differentiation among populations indicates that the management for the conservation of genetic variability in S. dolichocarpa should aim to preserve every population.     

Amplified fragment length polymorphism analysis of the population structure and genetic diversity of Phoebe zhennan (Lauraceae), a native species to China

Phoebe zhennan S. Lee et F. N. Wei (Lauraceae), is the main source of Gold Phoebe, a rare and extremely valuable wood in China. However it has undergone a dramatic decline. In this study, we used 12 amplified fragment length polymorphism primer combinations to assay 92 accessions, which were highly representative of the entire P. zhennan germplasm. It revealed that P. zhennan consisted of three genetic populations, named as SCZ (central Sichuan), CQH (eastern Sichuan, Chongqin, Hubei and Hunan) and YG (Yunnan and Guizhou), probably owing to natural selection caused by topography differences. The CQH population further diverged into two geographical sub-populations: CD-CQ (SCD and west region of Chongqin) and HB-HN (eastern side of Chongqin, Hubei and Hunan). The loci were moderately polymorphic (40.4%). The genetic distance between SCZ and YG was the highest, between CD-CQ and HB-HN the lowest. Pairwise fixation indices (Ф_PT) between any inferred populations were significant. This rare species exhibited low genetic diversity; therefore, the results provided significant data related to the conservation and management of P. zhennan. That is, with this genetic information, land managers are equipped with better tools allowing them to more effectively protect this species and its limited genetic diversity.     

Start Codon Targeted (SCoT) marker reveals genetic diversity of Dendrobium nobile Lindl., an endangered medicinal orchid species

Genetic variability in the wild genotypes of Dendrobium nobile Lindl. collected from different parts of Northeast India, was analyzed using a Start Codon Targeted (SCoT) marker system. A total of sixty individuals comprising of six natural populations were investigated for the existing natural genetic diversity. One hundred and thirty two (132) amplicons were produced by SCoT marker generating 96.21% polymorphism. The PIC value of the SCoT marker system was 0.78 and the Rp values of the primers ranged between 4.43 and 7.50. The percentage of polymorphic loci (Pp) ranging from 25% to 56.82%, Nei's gene diversity (h) from 0.08 to 0.15 with mean Nei's gene diversity of 0.28, and Shannon's information index (I) values ranging from 0.13 to 0.24 with an average value of 0.43 were recorded. The gene flow value (0.37) and the diversity among populations (0.57) demonstrated higher genetic variation among the populations. Analysis of molecular variance (AMOVA) showed 43.37% of variation within the populations, whereas 56.63% variation was recorded among the populations. Cluster analysis also reveals high genetic variation among the genotypes. Present investigation suggests the effectiveness of SCoT marker system to estimate the genetic diversity of D. nobile and that it can be seen as a preliminary point for future research on the population and evolutionary genetics of this endangered orchid species of medicinal importance.     

Genetic Variation and Differentiation of Larix decidua Populations in Swiss Alps (in English)

Genetic diversity within and among six subpopulations of Larix decidua Mill. from two altitudinal transects of Swiss Alps was investigated using 6 enzyme systems coding for 8 loci. Globally, the mean proportion of polymorphic loci was 22.9%, the average number of alleles per locus was 1.3, and the mean expected heterozygosity was 0.095. Only 5.8% of the genetic variation resided among populations. The mean genetic distance was 0.006. Several significant differences of gene frequencies were found between different age classes. Positive values of the species mean fixation index observed in this study suggested a considerable deficit of heterozygotes in the populations of L. decidua of Swiss Alps. At one of the sites (Arpette), the highest subpopulation in elevation gave the lowest level of genetic diversity (as evidenced by the lowest proportion of polymorphic loci and the lowest mean expected heterozygosity) and the largest value of genetic distance when compared to other subpopulations. The genetic differences between the highest subpopulation and the other ones suggest that the founder effect may be an important factor influencing genetic differentiation of L. decidua populations at Arpette transect.     

Genetic diversity within and among populations of Neopicrorhiza scrophulariiflora (Scrophulariaceae) in China,an endangered medicinal plant

Neopicrorhiza scrophulariiflora Pennel (Hong) (Scrophulariaceae) is an endangered medicinal plant, endemic to the Eastern Himalayas and the Hengduan Mountains. Levels of genetic variation and genetic structure of seven populations of N. scrophulariiflora in China were studied using inter-simple sequence repeat (ISSR) markers. Thirteen primers amplified 82 total loci from 7 populations composed of 136 individuals. All 82 loci were polymorphic, showing a percentage of polymorphic bands (PPB) of 100%, indicating considerable genetic variation at the species level. In contrast, a low level of genetic diversity within populations was detected with a mean PPB of 30.56%. In addition, Nei’s genetic diversity analysis (0.4073) and Shannon’s diversity index (0.5917) revealed similar genetic structure. High levels of genetic differentiation (Gst = 0.6955) and restricted gene flow (Nm = 0.2198) among populations were also detected. Anthropologic impacts, together with clonal propagation, genetic drift and geographical isolation might be the reasons which had shaped the genetic structure of this species.     

瑞士阿尔卑斯山欧洲落叶松居群的遗传变异和分化

应用等位酶分析技术,沿两个海拔梯度在瑞士阿尔卑斯山研究了欧洲落叶松（Larix decidua Mill.)居群内和居群间的遗传多态性,结果表明,多态性位点的比例为22.9%,平均每个位点的等位基因数为1.3,平均期望杂合度为0.095,遗传变异的5.8%存在于居群之间,平均遗传距离为0.006。几个有统计意义的基因频率差别在不同树龄类被发现,正在繁育系数值表明在瑞士阿尔卑斯山欧洲落叶松居群有相当的杂合体缺乏,在阿尔拜特（Ar-pette),海拔最高的亚居群与其他亚居群相比较显示最低的遗传多态性（如显示最低的我态性位点比例和最低的平均期望杂合度）和最大的遗传距离值,在最高亚居群和其他亚居群间的遗传差别暗示奠基效应可能是影响这个定样场所欧洲落叶松居群遗传分化的主要因素。     

白沙蒿不同地理种群遗传分化的ISSR分析

运用ISSR技术,对白沙蒿的5个种群进行遗传分化的分析。12个引物在108个个体中共扩增出222个位点,其中,多态位点为218个,多态位点百分率为98.20%,Shannon多样性指数为0.315 4,具有高的多态性。种群间的遗传分化系数（G_st）为0.076 7,与AMOVA分析的结果一致（Ф_st为7.96%）,表明绝大多数的遗传变异存在于种群内部。各种群间遗传一致度高达98%以上,遗传距离很小,基因流达3.008 2,均表现出白沙蒿各种群存在着广泛的基因交流,有着很小的遗传分化。     

Variation in the sequence of a mitochondrial nadh dehydrogenase i gene fragment among six natural populations of Schistosoma japonicum from China

The present study investigated the level of genetic variation among Schistosoma japonicum populations of different geographical origins from mainland China. Polymerase chain reaction-based methods were employed to determine the sequence for a subunit of the mitochondrial NADH dehydrogenase I gene for populations from Zhejiang, Anhui, Jiangxi, Hunan, Hubei and Sichuan. No variation was detectable in the NADH dehydrogenase I sequence within populations from Zhejiang and Hubei, whereas sequence variation of 0.2% was detected within populations from Anhui, Jiangxi, Hunan and Sichuan. Pairwise comparison of the sequences representing the six different populations revealed genetic differences ranging from 0 to 0.6%.     

Genetic variation and population structure ofCarex breviculmis (Cyperaceae) in Korea

We determined the genetic diversity and population structures ofCarex breviculmis (Cyperaceae) populations in Korea, using genetic variations at 23 allozyme loci.C. breviculmis is a long-lived herbaceous species that is widely distributed in eastern Asia. A high level of genetic variation was found in 15 populations. Twelve enzymes revealed 23 loci, of which 11 were polymorphic (47.8%). Genetic diversity at the speciesand population levels were 0.174 and 0.146, respectively. Total genetic diversity (H_T = 0.363) and within-population genetic diversity (H_s = 0.346) were high, whereas the extent of the population divergence was relatively low (G_ST = 0.063). Deviation from random mating (Fis) within the 15 populations was 0.206. An indirect estimate of the number of migrants per generation(Nm = 3.69) indicated that gene flow was extensive among Korean populations of this species. Analysis of fixation indices revealed a substantial heterozygote deficiency in some populations and at some loci. Genetic identity between popu-lations was high, exceeding 0.956.     

大别山山核桃天然群体遗传结构的初步分析

利用RAPD分子标记技术检测了大别山山核桃3个天然居群的遗传多样性和遗传结构。20条10 bp随机引物共检测到238个扩增位点,其中多态性位点162个,多态位点百分率(PPB)为68.1%。居群水平Shannon’s多态性信息指数(I)介于0.2651~0.2801之间;居群水平Ne i’s基因多样性指数(H)介于0.1789~0.1890之间。遗传变异计算显示大别山山核桃居群间基因分化系数(Gst)为0.4063,分子方差分析(AMOVA)表明居群间基因分化水平为0.4177,居群间基因流(Nm)为0.7306,说明大别山山核桃大部分变异存在于居群内,居群间基因交流相对较少。这一结果符合大别山山核桃风媒、异交的繁育系统特点,但其居群间基因分化程度明显高于异交植物的平均水平(Gst=0.1930)。地理隔离、居群内近交及居群间基因流受阻可能是形成目前大别山山核桃天然群体遗传结构的主要因素。     

Genetic diversity and biogeography of the traditional Chinese medicine, Gardenia jasminoides, based on AFLP markers

Gardenia jasminoides Ellis is used in traditional Chinese medicine (TCM) in China. Levels of genetic variation and patterns of population structure within and among eight wild or cultivated populations of G. jasminoides Ellis in China were investigated using amplified fragment length polymorphism (AFLP) markers. Of the 11 primers screened, four produced highly reproducible AFLP bands. Using these primers, 244 discernible DNA fragments were generated with 165 bands (67.6%), were polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 36.89% to 59.43%. Genetic diversity within populations ranged from 0.2086 to 0.3108, averaging 0.2392 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (76.59%), Shannon's index analysis (64.8%) and AMOVA analysis (72.75%). No significant statistical differences (analysis of molecular variance [AMOVA], p = 0.0639) in AFLP variation were found between regions. However, the variance among populations and within populations differed significantly (p < 0.001). An indirect estimate of historical levels of gene flow (N_m = 1.7448) was consistent with the high mean genetic identity (mean I = 0.9263) found among populations. There is an association between geographic and genetic distances between populations. Presently gene change exists between populations.