Genetic diversity of forest arabica coffee (Coffea arabica L.) in Ethiopia as revealed by random amplified polymorphic DNA (RAPD) analysis

Genetic diversity within the forest Coffea arabica L. gene pool in Ethiopia has not been extensively examined with molecular markers. In the present study, a total of 75 polymorphic RAPD bands generated by twelve random primers were used to assess genetic diversity among 144 genotypes representing 16 C. arabica populations. The number of polymorphic bands detected with each primer ranged from 2 to 9 with a mean of 6.25 bands per primer. Banding patterns ranged in percentage polymorphism from 37% to 73% with an overall mean of 56% for the populations analyzed. The amount of genetic variation among populations estimated by Shannon-Weaver diversity index was (H = 0.30). The within population and between populations differentiation values were 0.65 and 0.35, respectively. Genetic differentiations within and between zones of sample collection sites were 0.80 and 0.20, respectively. Within population average similarities estimated by simple matching coefficients ranged from 0.72 to 0.85, with an overall average of 0.78. In the cluster analysis that used individual samples as operational taxonomic units, most of the representatives of the same population failed to cluster before they joined members of other populations. Nevertheless, most of the populations were clustered on the basis of their geographic closeness and an east west differentiation was observed at approximately 75% similarity. The results obtained provide information on how to select sites for in situ conservation of C. arabica germplasm.     

Randomly amplified polymorphic DNA-polymerase chain reaction analysis of two different populations of cultured Korean catfishSilurus asotus

Genetic similarity and diversity of cultured catfishSilurus asotus populations collected from two areas in western Korea were examined using randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Out of 20 random primers tested, 5 produced 1344 RAPD bands ranging from 8.2 to 13.6 polymorphic bands per primer. The polymorphic bands in these populations ranged from 56.4% to 59.6%. Polymorphic bands per lane within populations ranged from 4.9% to 5.3%. The similarity within the Kunsan population varied from 0.39 to 0.82 with a mean (± SD) of 0.56 ± 0.08. The level of bandsharing values was 0.59 ± 007 within the catfish population from Yesan. The genetic similarity in cultured catfish populations may have been caused because individuals from two populations were reared in the same environmental conditions or by inbreeding during several generations. However, in view of bandsharing values, polymorphic bands and also the specific major bands that were inter-population-specific, significant genetic differentiation between these populations were present even if bandsharing (BS) values were somewhat numerically different. Therefore, the number of RAPD polymorphisms identified in this study may be sufficient to permit estimating genetic similarity and diversity. However, in future, additional populations, sampling sites and individuals will be necessary to make up for these weak points.     

Microsatellite variability and heterozygote deficiency in the arctic-alpine Alaskan wheatgrass (Elymus alaskanus) complex.

Genetic variation in the allotetraploid grass Elymus alaskanus complex was assessed using microsatellites in seven populations from Canada, Greenland, and the U.S.A. Microsatellite variation was compared with allozyme and RAPD variation. Our results indicated that E. alaskanus was highly homozygous but also highly variable. The polymorphic loci ranged from 50 to 100% with a mean of 78.6%, and the mean number of allele per locus was 3.14. Average expected heterozygosity value (HE, gene diversity) varied across populations and ranged from 0.244 to 0.651 with mean of 0.414. The mean value of HE across Canadian populations (0.517) was significantly higher than that across populations in Greenland (0.367). The correlation between allozyme and microsatellite gene diversity value (HE) showed a high positive correlation (r = 0.68), but between RAPD and microsatellite showed a low positive correlation (r = 0.08). Populations were highly differentiated, with 38% of variation among populations. Interpopulation genetic distance showed no association with geographic distance between the population sites of origin. A Hardy-Weinberg exact test for all loci and all populations reveals a significant heterozygote deficiency. Possible explanations for heterozygote deficiency are discussed.     

AFLP and RAPD analyses of genetic diversity of wild and/or weedy Guizotia (Asteraceae) from Ethiopia

Amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers were used to provide estimates of the comparative genetic variation within and among populations of various Guizotia taxa with the goal of conserving and utilizing their genetic diversity. The percentage of polymorphic loci (P(S)) ranged from 28.5%-90% (AFLP) and 85.6%-99.6% (RAPD). The overall gene diversity estimate () has shown slight variation among taxa ranging from 0.32-0.37 (AFLP) and from 0.22 to 0.28 (RAPD). The within population diversity of "Chelelu" and "Ketcha" was found to be unexpectedly high. Both parameters used to estimate population differentiation (G(ST) and F(ST)) revealed the highest population differentiation G. zavattarii in followed by G. arborescens. Genetic variation among populations within a taxon was highly significant for all the five taxa as revealed by AMOVA (P<0.0001). The need for immediate conservation activities for G. arborescens and G. zavattarii, and factors that contribute to the existing genetic variability and population genetic structures are discussed.     

Microsatellite analysis of genetic diversity and population genetic structure of a wild rice (<Emphasis Type="Italic">Oryza rufipogon</Emphasis> Griff.) in China

Genetic diversity and population genetic structure of natural Oryza rufipogon populations in China were studied based on ten microsatellite loci. For a total of 237 individuals of 12 populations collected from four regions, a moderate to high level of genetic diversity was observed at population levels with the number of alleles per locus ( A) ranging from 2 to 18 (average 10.6), and polymorphic loci ( P) from 40.0% to 100% (average 83.3%). The observed heterozygosity ( H(O)) varied from 0.163 to 0.550 with the mean of 0.332, and the expected heterozygosity ( H(E)) from 0.164 to 0.648 with the mean of 0.413. The level of genetic diversity for Guangxi was the highest. These results are in good agreement with previous allozyme and RAPD studies. However, it was unexpected that high genetic differentiation among populations was found ( R(ST) = 0.5199, theta = 0.491), suggesting that about one-half of the genetic variation existed between the populations. Differentiation (pairwise theta) was positively correlated with geographical distance ( r = 0.464), as expected under the isolation by distance model. The habitat destruction and degradation throughout the geographic range of O. rufipogon may be the main factor attributed to high genetic differentiation among populations of O. rufipogon in China.     

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.     

Genetic Diversity and Genetic Structure of an Endangered Species, Trillium tschonoskii

The genetic diversity and genetic structure of Trillium tschonoskii (Maxim) were investigated using amplified fragment length polymorphism markers. Eight primer combinations were carried out on 105 different individuals sampled from seven populations. Of the 619 discernible DNA fragments generated, 169 (27.3%) were polymorphic. The percentage of polymorphic bands within populations ranged from 4.52 to 10.50. Genetic diversity (H_E) within populations ranged from 0.0130 to 0.0379, averaging 0.0536 at the species level. Genetic differentiation among populations was detected based on Nei's genetic diversity analysis (53.03%) and analysis of molecular variance (AMOVA) (52.43%). AMOVA indicated significant genetic differentiation among populations (52.43% of the variance) and within populations (47.57% of the variance) (p < 0.0002). Gene flow was low (0.4429) among populations. Species breeding system and limited gene flow among populations are plausible reasons for the high genetic differentiation observed for this species. We propose an appropriate strategy for conserving the genetic resources of T. tschonoskii in China.     

Genetic diversity of the tropical tree Terminalia amazonia (Combretaceae) in naturally fragmented populations

The effect of long-term fragmentation on the genetic diversity of populations of the neotropical tree species, Terminalia amazonia, was studied using random amplified polymorphic DNA (RAPD) analysis. Leaf material from 104 trees was collected from three naturally fragmented gallery forest patches and three plots in nearby continuous forest in the Mountain Pine Ridge, Belize. In total, 30 RAPD bands generated by five decamer primers were used to compare the genetic diversity of the six populations in the two groups. Genetic variation within the populations (H0), as estimated by the Shannon diversity index, ranged from 0.32 to 0.38, with an overall diversity of 0.38 (Hspecies). Analysis of molecular variation revealed that most (94.4%, P<0.001) of the variation was attributable to differences among individuals within populations. Population differentiation was significantly (P=0.038) lower among the fragmented populations than among continuous forest populations. On average, the fragmented populations also had slightly, but statistically significant (P=0.046) lower levels of genetic diversity. However, one gallery forest site had a higher level of genetic diversity than two of the continuous forest sites. We suggest that the long-term effect of fragmentation on the genetic diversity of tropical trees will depend upon the amount of local forest cover in proximity to the fragmented populations.     

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.     

栲树天然群体遗传结构的RAPD分析

利用RAPD分子标记对 5个栲树 (CastanopsisfargesiiFranch .)天然群体共计 188个个体的遗传多样性和群体遗传结构进行了分析。 4 1个随机寡核苷酸引物共检测到 385个位点 ,其中多态位点 15 7个 ,占 4 0 .78%。物种水平的Shannon多样性指数I=0 .4 5 97,Nei基因多样度h =0 .2 96。遗传变异分析表明 ,栲树群体的遗传变异主要存在于群体内 ,利用Shannon多样性指数估算的分化 (Hsp_Hpop) /Hsp=0 .0 4 76 ,遗传分化系数Gst =0 .0 4 2 9,分子方差分析 (AMOVA)也证实了这一结论 ,群体内的变异组分占了 94 .97% ,群体间变异只占 5 .0 3%。AMOVA分析结果的显著性检验也表明 ,群体间及群体内个体间均呈现出显著分化 (P <0 .0 0 1)。     

利用AFLP分子标记探讨蜡梅种质资源的遗传多样性

利用AFLP分子标记技术,对中国蜡梅种质资源7个野生种群的遗传多样性进行了研究。利用筛选出的3对引物,共扩增出253条谱带,其中218条多态带,多态位点占86.17% ;种群间的基因分化系数为0.2906,说明蜡梅基因多样性主要存在于种群内;种群总的Nei s基因多样性指数为0.2933,Shannon信息多态性指数为0.4487,蜡梅总的遗传多样性水平较高。蜡梅不同种群遗传多样性水平差异较大,种群多态位点百分率在65.44% ～87.16%之间,Nei s基因多样性指数为0.1653 ～0.4012,Shannon信息多态性指数为0.3132 ～0.5603。神农架种群(SN)和保康种群(BK)的遗传多样性水平较高。用NTSYS2.01版软件对样品进行UPGMA聚类分析,结果7个种群并没有按地理距离进行聚类。最后提出要对各蜡梅野生群体采取相应的迁地和就地保护措施。     

中华水韭遗传多样性的RAPD分析

采用RAPD方法对珍稀濒危植物中华水韭(Isoetes sinensis)4个自然居群的48个样品进行了DNA多态性分析。从60个随机引物中筛选出14个有效引物,共产生124条DNA片段,其中72条为多态性条带,总的多态位点百分率(PPB)为58．06％。各居群间多态位点百分率差异显著(0．81％-12．90％)。AMOVA分析结果表明,4个居群间基因分化系数Фst=0．5894,即遗传变异中有相当一部分来源于群体间(58．94％)。日益缩小的种群规模而导致的居群内近交和遗传漂变的发生以及居群间有限的基因交流可能是中华水韭目前遗传结构的主要成因。鉴于目前中华水韭居群内个体数偏少、遗传多样性较低的现状,建议对其进行就地保护并保护尽可能多的生境,对不同自然居群内的个体进行植株相互移栽和育苗移栽,以提高不同居群间的基因交流,尽可能地保护中华水韭的遗传多样性。     

Genetic diversity analysis of Rhododendron aureum Georgi (Ericaceae) located on Changbai Mountain using ISSR and RAPD markers

Rhododendron aureum Georgi (Ericaceae) is a perennial alpine shrub endemic to Changbai Mountain in China. We used ISSR and RAPD markers to describe the diversity and genetic structure within and among four natural populations located at different altitudes. DNA from 66 individuals was amplified with ten ISSR markers and seven RAPD markers. High genetic diversity was observed by these two techniques at the species level. The genetic diversity of populations increased with altitudinal gradients from low to high. The coefficient of gene differentiation (G_ST 0.3652 in ISSR and 0.2511 in RAPD) and AMOVA analysis revealed that most genetic diversity was distributed within populations (61.96% in ISSR and 70.23% in RAPD). The estimate of gene flow based on G_ST was 0.8690 in ISSR and 1.4910 in RAPD. The UPGMA clustering results using ISSR and RAPD showed that all individuals from the same altitude were gathered together, and the two populations (TYD2a and YHLa) from middle altitudes always clustered together. Compared with populations from different altitudes, similar genetic diversity and low genetic differentiation were obtained from populations at the same altitudes, as revealed by ISSR markers. In addition to the reproductive strategy of R. aureum, these data highlight that local environmental conditions may play an important role in shaping the diversity and genetic structure of this species.     

我国部分金莲花种质资源遗传多样性的RAPD研究

用RAPD标记对来自不同产地的24份金莲花种质从分子水平开展遗传多样性研究,为综舍评价我国金莲花资源现状及植物资源的合理保护提供参考。结果显示,30条RAPD引物共扩增获得194条清晰可辨条带,其中多态性条带108条,多态性位点百分率为55．67％,金莲花总多样性指数为0．1460,居群内遗传多样性为0．0591,Shannon’s多样性指数为0．2248;通过UPGMA进行聚类分析,将24份金莲花种质划分为3个大类,根据总遗传多样性和居群内遗传多样性计算不同居群间遗传分化系数为0．6902,基因流估计平均值为0．2244。研究结果表明,目前我国金莲花种质资源的遗传多样性指教偏低,应该尽快采取相应措施,对不同金莲花资源进行合理保护。     

天然红松群体遗传多样性的RAPD分析

用RAPD技术分析了分布于中国东北的3个红松（Pinus koraiensis Seib.et Zucc.）天然群体的遗传多样性及群体间的遗传分化。38个随机引物共检测到241个可重复的位点,其中多态位点139个,占总位点的57.68%。Shannon信息指数和Nei指数的统计结果都表明,红松种内的遗传变异主要存在于群体内,凉水群体的遗传多样性水平高于黑河、虎林群体。群体内遗传相似度为0.927,群体间为0.845。红松现阶段对偏低的遗传多样性水平与第四纪冰期所遭受的严重打击和人类近期的干扰有较大关系。     

Genetic variability and differentiation of Caragana microphylla populations as revealed by RAPD markers

Genetic variability in random amplified polymorphic DNA (RAPD) was studied in 90 individuals of Caragana microphylla, an outcrossing perennial shrub species, from five natural populations sampled in Inner Mongolia steppe of China on a small scale. Nineteen selected primers were used to amplify DNA samples, and totally 225 bands were detected. The percentage of polymorphic bands within populations ranged form 58.22% to 63.56%, with an average of 60% at the population level and 71.11% at the species level, indicating relatively high genetic variations in C. microphylla species. Shannon's information index (I) and Nei's gene diversity (h) showed the similar trend with each other. According to the analysis of Nei's gene diversity, the percentage of genetic variation among populations was 7.13%, indicating a low level of genetic differentiation among populations. There existed a strong gene flow (Nm = 3.26) among populations. Although AMOVA analysis also revealed most variation was within populations (phi(ST) = 4.1%), a significant proportion was observed among populations (P<0.001) in the present study, suggesting genetic differentiation occurred among populations at a certain extent. Based on Mantel's tests and the results of previous studies, the genetic structure pattern of C. microphylla accorded with the isolation-by-distance model on a very large scale, however, on a small scale, the significant genetic differentiation among populations might be enhanced by the micro-environmental divergence among the sampling sites, rather than by geographic factors. Analysis of the genetic variations of C. microphylla populations provided useful information for the adaptive strategy of Caragana species.     

Comparative genetic diversity of parasites and their hosts: population structure of an urban cockroach and its haplo-diploid parasite (oxyuroid nematode)

Few studies have investigated the genetic structure of both host and parasite populations at a level of populations and at a level of individuals. We investigated the genetic structure of the urban cockroach Blattella germanica and its oxyuroid parasite Blatticola blattae. Random amplified polymorphic DNA (RAPD) markers were used to quantify genetic diversity between and within four populations (from two cities in France) of the host and its parasite. Diversity based on phenotypic frequencies was calculated for each RAPD marker using Shannon-Wiener's index. We used multivariate analyses to test the significance of genetic differentiation between host and parasite populations. Analysis of molecular variance was also used. Both methods gave similar results. Diversity between pairs of individuals was estimated by Nei & Li's index. Genetic diversity was higher within host or parasite populations (80% and 82%, respectively, of explained diversity) than between host or parasite populations (20% and 18%, respectively, explained diversity). The genetic distances between pairs of parasite populations (or individuals) were not correlated with the genetic distances between the corresponding pairs of host populations (or individuals).     

RAPD analysis of genetic diversity and population structure of Elymus sibiricus (Poaceae) native to the southeastern Qinghai-Tibet Plateau, China

Genetic diversity of Elymus sibiricus (Poaceae) was examined in eight populations from the southeast Qinghai-Tibet Plateau. We detected 291 RAPD polymorphic loci in 93 samples. The percentage of polymorphic bands (PPB) was 79%. Genetic diversity (H(E)) was 0.264, effective number of alleles (N(E)) was 1.444, Shannon's information index (H(O)) was 0.398, and expected Bayesian heterozygosity (H(B)) was 0.371. At the population level, PPB = 51%, N(E) = 1.306, H(E) = 0.176, I = 0.263, and H(B) = 0.247. A high level of genetic differentiation was detected based on Nei's genetic diversity analysis (G(ST) = 32.0%), Shannon's index analysis (33.7%), and the Bayesian method (θ(B) = 33.5%). The partitioning of molecular variance by AMOVA demonstrated significant genetic differentiation within populations (60%) and among populations (40%). The average number of individuals exchanged between populations per generation (N(m)) was 1.06. The populations were found to share high levels of genetic identity. No significant correlation was found between geographic distance and pairwise genetic distance (r = 0.7539, P = 0.9996). Correlation analysis revealed a significant correlation (r = 0.762) between RAPD H(E) found in this study and ISSR H(E) values from a previous study.     

羊草种群遗传分化的RAPD分析Ⅱ.RAPD数据的统计分析

对松嫩草原上分布的灰绿型和黄绿型羊草9个种群进行了15个引物的RAPD分析,统计结果表明,两类种群的扩增片段数和多态位点比率明显不同,黄绿型种群低于灰绿型,其值分别＜90与＞100,＜50％与＞70％,比较了7种不同统计方法据RAPD表型或基因型频率估算的种群遗传多样性,几种统计结果都揭示,黄绿型种群低于灰绿型种群,用F1s值矫正种群对Hardy-Weinberg平衡的偏离后,估算等位基因频率,通过Shannon指数和Nei指数估计羊草种群间分化分别为37．6％和35．7％,高于等位酶的分析,讨论比较了等位酶和RAPD分析结果的异同。     

Genetic diversity in Hemileia vastatrix based on RAPD markers

Random amplified polymorphic DNA (RAPD) was used to assess the genetic structure of Hemileia vastatrix populations. Forty-five rust isolates with different virulence spectra and from different hosts and geographical regions were analyzed. Out of 45 bands, generated with three RAPD primers, 35 (78%) were polymorphic and scored as molecular markers. Cluster analysis exhibits unstructured variability of this pathogen with regard to physiological race, geographical origin or host. The genotypic diversity (H') inferred from Shannon's index was higher than gene diversity (Ht), suggesting that diversity is distributed among clonal lineages. Estimates of gene diversity in Africa and Asia populations were higher in total (Ht) as compared to within population diversity (Hs). Genetic differentiation was considerable among coffee rust isolates from Africa (Gst = 0.865) and Asia (Gst = 0.768) but not among isolates from South America (Gst = 0.266). We concluded that genetic diversity in H. vastatrix was moderately low and that the genetic differentiation among populations shows that asexual reproduction is likely to play an important role in the population biology of this fungus. This should be taken into account for the development of breeding programs.