Genetic diversity of <Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass. (Asteraceae) from Ethiopia as revealed by random amplified polymorphic DNA (RAPD)

Genetic diversity of 70 populations of niger (Guizotia abyssinica) representing all its growing regions in Ethiopia was investigated using random amplified polymorphic DNA (RAPD) to reveal the extent of its populations genetic diversity. Ninety-seven percent of the loci studied was revealed to be polymorphic for the whole data set. The within population diversity estimated by Shannon diversity index and Nei gene diversity estimates was revealed to be 0.395 and 0.158, respectively. The extent of genetic variation of populations from major niger producing regions was significantly lower than that of populations from other regions; however, it is distributed regardless of altitude of growth. Genetic differentiation between populations was estimated with Shannon index as G^′ _ST (0.432), Nei’s G _ST (0.242) and AMOVA based F _ST (0.350) and appears to be equivalent to the average values calculated from various RAPD based studies on outcrossing species. Higher proportion of the variation detected by AMOVA resided within populations (64.58%) relative to the amount of variation among populations (35.42%). UPGMA cluster analysis showed that most of the populations were clustered according to their region of origin. However, some populations were genetically distant from the majority and seem to have unique genetic properties. It is concluded that the crop has a wide genetic basis that may be used for the improvement of the species through conventional breeding and/or marker assisted selection. Collection of germplasm from areas not yet covered and/or underrepresented is the opportunity to broaden the genetic basis of genebank collection.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Study of clonally propagated cassumunar ginger (<Emphasis Type="Italic">Zingiber montanum</Emphasis> (Koenig) Link ex Dietr.) and its relation of wild <Emphasis Type="Italic">Zingiber</Emphasis> species from Thailand revealed by RAPD markers

The genetic relatedness among 51 accessions, 14 species of the genus Zingiber and genetic variability of a clonally propagated species, Zingiber montanum (Koenig) Link ex Dietr., from Thailand were studied using random amplified polymorphic DNA (RAPD) profiles. Twenty-nine random primers gave reproducible amplification banding patterns of 607 polymorphic bands out of 611 scored bands accounting for 99.40% polymorphism across the genotypes. Jaccard’s coefficient of similarity varied from 0.119 to 0.970, indicative of distant genetic relatedness among the genotype studied. UPGMA clustering indicated eight distinct clusters of Zingiber, with a high cophenetic correlation (r = 1.00) value. Genetic variability in Z. montanum was exhibited by the collections from six regions of Thailand. High molecular variance (87%) within collection regions of Z. montanum accessions was displayed by AMOVA and also explained the significant divergence among the sample from six collection regions. Our results indicate that RAPD technique is useful for detecting the genetic relatedness within and among species of Zingiber and that high diversity exists in the clonally propagated species, Z. montanum.     

Genetic Structure of Wild and Domesticated Populations of Capsicum annuum (Solanaceae) from Northwestern Mexico Analyzed by RAPDs

Levels of genetic variation and genetic structure of 15 wild populations and three domesticated populations of Capsicum annuum were studied by RAPD markers. A total of 166 bands (all of them polymorphic) and 126 bands (125 of them polymorphic) were amplified in wild and domesticated populations, respectively. Mean percentage of polymorphism was 34.2% in wild populations and 34.7% in domesticated populations. Mean and total genetic diversity were 0.069 and 0.165 for wild populations and 0.081 and 0.131 for domesticated populations. Parameters of genetic diversity estimated from 54 bands with frequencies ≥1 − (3/n) (n = sample size) showed that 56.7% of the total variation was within and 43.3% among wild populations, whereas 67.8% of the variation was within and 32.2% among domesticated populations. AMOVA indicated that total genetic diversity was equally distributed within (48.9 and 50.0%) and among (50.0 and 51.1%) populations in both wild and domesticated samples. Wild and domesticated populations were clearly resolved in a UPGMA dendrogram constructed from Jaccard’s distances (average GD = 0.197), as well as by AMOVA (17.2% of variance among populations types, p = 0.001) and by multidimensional scaling analysis. Such differentiation can be associated with domestication as well as different origin of gene pools of the wild (Northwestern Mexico) and cultivated (more probably Central Mexico) samples analyzed. The considerable genetic distances among cultivars (average GD = 0.254) as well as the high number of diagnostic bands per cultivar (33 out of 126 bands), suggest that genetic changes associated with domestication could have resulted from artificial selection intervening in different directions, but the inclusion of more domesticated samples might clarify the nature of distinctions detected here.     

Genetic Diversity Among Brazilian Cultivars and Landraces of Tomato Lycopersicon Esculentum Mill. Revealed by RAPD Markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

Abundant genetic diversity in cultivated <Emphasis Type="Italic">Codonopsis pilosula</Emphasis> populations revealed by RAPD polymorphisms

Genetic diversity of seven cultivated populations of Codonopsis pilosula Nannf. from Longxi County, Gansu Province of China was estimated using randomly amplified polymorphic DNA (RAPD) markers. The 17 selected RAPD primers amplified 205 polymorphic bands out of a total of 235 (87.2%). Nei’s gene-diversity statistics and population differentiation parameters based on AMOVA analysis indicated that the cultivated C. pilosula populations remained a high level of genetic diversity with Hs = 0.299 and I = 0.450. A greater proportion of genetic diversity was found within (77%) rather than among (23%) the populations. In addition, we also detected that populations from different altitudes had a considerable genetic differentiation after 40 years of cultivation at the same site. Populations from higher altitude had lower genetic diversity than those from lower altitude. Our results suggested that irregular and sparse cultivation practices, i.e., random collecting, preserving, and planting seeds of the medicinal species without deliberate selection, might be an efficient way to conserve genetic resources of medicinal plants, in addition to their effective uses.     

An Assessment of the Genetic Diversity within a Collection of Wild Cranberry (<Emphasis Type="Italic">Vaccinium macrocarpon</Emphasis> Ait.) Clones with RAPD-PCR

Forty-three wild cranberry (Vaccinium macrocarpon Ait.) clones collected from four Canadian provinces and five cranberry cultivars were assessed for genetic variability by using random amplified polymorphic DNA (RAPD)-PCR. Fourteen primers generated 161 polymorphic RAPD-PCR bands. A substantial degree of genetic diversity was found among the wild cranberry collections. Cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) separated the wild clones and three cultivars into five main clusters, and identified the two remaining cultivars as outliers. Furthermore, within four clusters, the genotypes tended to form sub-clusters that were in agreement with the principal coordinate (PCO) analysis. Geographical distribution explained 10% of total variation as revealed by analysis of molecular variance (AMOVA). The RAPD markers detected a sufficient degree of polymorphism to differentiate among cranberry clones and cultivars, making this technology valuable for germplasm management and the more efficient choice of parents in current cranberry breeding programs.     

Population genetic structure and the conservation of isolated populations of Acacia raddiana in the Negev Desert

There is much concern over the high mortality of many populations of Acacia raddiana, a keystone tree species in the Negev desert of Israel. We used random amplified polymorphic DNA (RAPD) to assess patterns of genetic variation within and among 12 populations of A. raddiana from the Arava (Syrian-African Rift) valley and western Negev. A high level of genetic polymorphism was recorded within populations. An analysis of molecular variance (AMOVA) showed that about 59.4% of total genetic variance occurred among populations, which is considerably greater population differentiation than that recorded for other outbreeding species. Cluster and principal coordinates analyses and AMOVA indicate that the western Negev and Arava valley populations are highly differentiated. We suggest that there may have been two invasions of A. raddiana into Israel: one across the northern Sinai/Gaza Strip area into the western Negev, with some plants reaching the Dead Sea and a second invasion across the southern part of the Sinai peninsula, or even from Saudi Arabia, up to the Arava valley. From the conservation point of view, each population should be conserved separately because they are genetically highly differentiated and loss of any one population would lead to a dramatic loss of genetic variation. The mixing of genetically distinct populations may give rise to outbreeding depression (particularly because of GXE interactions). An obvious first step to the maintenance of this species' genetic diversity is the separate management of the western Negev and Arava valley populations because of their different evolutionary histories.     

Conservation genetics and implications for restoration of Hemigenia exilis (Lamiaceae), a serpentine endemic from Western Australia

Hemigenia exilis (Lamiaceae) is a rare plant endemic to serpentine soils of the Goldfields of Western Australia. The species was presumed extinct until 1995, when it was re-discovered on a nickel ore deposit. To delineate the origin and extent of seed collection for rehabilitation after mining, and to identify the impact of removing one population due to mining, we assessed level and partitioning of genetic variation and differentiation. Twelve populations were sampled for DNA fingerprinting using the random amplified polymorphic DNA (RAPD) technique. Ten primers produced 89 bands, 97% being polymorphic. Genetic diversity within populations ranged from 0.197 to 0.409, averaging 0.38 at the species level, which is high compared with most other endemic species. Heterozygosity within populations ranged from 0.355 to 0.431, averaging 0.27 over the species. AMOVA partitioned over 80% of the total variation within populations. Multidimensional scaling revealed weak but significant differentiation into a northern and southern provenance. Despite selective sampling, the genetic data provided useful information for the management of Hemigenia exilis. For restoration, seed should be collected from a range of habitats of several populations, while keeping the two provenances separate. This strategy is likely to maintain high genetic diversity and locally adapted populations.     

Genetic variation in the threatened South American conifer Pilgerodendron uviferum (Cupressaceae), detected using RAPD markers

Pilgerodendron uviferum (Cipres de las Guiatecas, Cupressaceae) is a long-lived conifer, endemic to southern Chile and Argentina, reaching a southern limit in Tierra del Fuego. Remnant populations are generally fragmented and highly disturbed because of exploitation for timber, grazing and fire. The extent of genetic variation within and between 16 populations of this species, distributed throughout its range, was assessed using random amplified polymorphic DNA (RAPD) markers. Eight 10-mer primers produced a total of 84 scorable markers, 30 of which (35.7%) were polymorphic. AMOVA indicated that 18.6% of the variation recorded was attributable to differences between populations, a relatively high value compared with other conifers from the region. Pair-wise Phi_st comparisons between populations were all significant at P<0.05, with one exception, highlighting a high degree of population differentiation. Values of Shannon's diversity index (S) differed significantly among populations (P=0.002, ANOVA), values ranging from 0.337 to 0.716, suggesting that some populations are currently characterised by very low genetic variability. Current patterns of genetic variation were related to biogeographic history and human impact. The high degree of population differentiation recorded here highlights the need for additional conservation measures for this species, both in terms of incorporating further populations into the protected areas, and the restoration of severely degraded populations, to ensure their continued viability.     

Population genetic structure in Lens taxa revealed by isozyme and RAPD analysis

An understanding of the genetic structure of populations is vital for the formation of optimum collection, conservation and utilization strategies for plant genetic resources. This is of particular importance in the case of in-situ conservation, a strategy gaining in popularity. The population genetic structures of five wild lentil taxa, Lens culinaris subsp. orientalis, L. odemensis, L. ervoides, L. nigricans and L. lamottei were investigated using isozyme electrophoresis and random amplified polymorphic DNA (RAPD). Approximately 20 plants from each of 5 populations per taxon were screened for variation at 11 isozyme loci and using three RAPD primers. Levels of variation were generally low, although considerable variation existed in the levels of diversity found within populations of L. culinaris subsp. orientalis and L. lamottei. Comparison of the results obtained in this study with the results obtained in a previous study indicate that this is a trend occurring across all species. It implies that levels of diversity within populations must be measured and considered prior to targeting of specific populations for in-situ conservation. Analysis of molecular variance of both isozyme and RAPD data revealed that between 78% and 99% of the variation was attributable to between-population differences. Isozyme results from L. lamottei populations were, however, contradictory. Possible explanations for this difference are discussed.     

Genetic differentiation in the olive complex (Olea europaea) revealed by RAPDs and RFLPs in the rRNA genes

We assessed the genetic differentiation of the Mediterranean olive from its wild relatives found in different geographic areas (Mediterranean, Asia, Africa) using eighty RAPDs revealed with eight primers. Variance analysis (AMOVA) enabled us to estimate the overall genetic differentiation parameters between wild populations. Oleasters from the Near East and Turkey were discriminated from the other Mediterranean populations. Olea laperrinei, O. maroccana and O. cerasiformis were the taxa the most related to the Mediterranean olive. In contrast, O. africana was shown to be the most genetically distant taxa from the Mediterranean olive. However, we characterised hybrid trees between these two taxa. Significant trends between genetic and geographic distances were met within the subspecies cuspidata and within the Mediterranean olive. A genetic diversity gradient was observed in both subspecies europaea and cuspidata. These results are in agreement with a mechanism of differentiation by distance in the O. europaea complex, but another non-exclusive mechanism could also be gene flow between differentiated taxa. Furthermore, we characterised the discriminating power of each RAPD to recognise the different taxa using intraclass correlation coefficients. Lastly, IGS-RFLPs enabled us to assess rDNA polymorphisms on a sub-sample of individuals. On the basis of these data, a low interspecifc differentiation was found. This suggests a recent genetic divergence between the different taxa of the O. europaea complex or the occurrence of gene flow during favourable periods or because human displacements. All the olive cultivars were genetically related to the oleaster populations supporting that Mediterranean is the olive domestication area.     

应用RAPD分析川西北高原老芒麦自然居群的遗传多样性

利用RAPD标记对来自青藏高原东南部川西北高原的8个老芒麦自然居群的遗传多样性和群体遗传结构进行了分析和评价。从150个RAPD引物中筛选出25个能扩增出高度重复性条带的引物。这25个引物共扩增出370条可分辨的条带,其中291条(占78.65%) 具有多态性,表明供试居群在物种水平上存在较高水平的变异。同时各居群的多态性位点比率(PP)在46.49%到53.78%之间变化,表明群体水平的变异较低。居群的平均基因多样性(HE)为0.176(变幅为0.159~0.190),而物种水平的平均基因多样性达0.264。基于Nei’s基因多样性、Shannon指数和贝叶斯方法的群体分化系数分别为32.0%、33.7%和33.5%。AMOVA 分析表明居群内遗传达到总变异的59.9%,而居群间变异仅有40.1%,但二者均达到极显著水平(P < 0.001)。居群间每世代迁入个体数(Nm)达到0.503个。各居群间存在较高的Nei’s遗传一致度。本研究获得的老芒麦的遗传结构不同于已报导的大多数披碱草属物种。另外,基于聚类分析及AMOVA的结果均表明各居群间存在较为明显的地理分化,8个居群分化为采集地的南部和北部2个分支。总之,研究结果表明来自青藏高原东南部的老芒麦居群具有较高水平的遗传变异。在该地区应尽量选择遗传多样性高的老芒麦居群实施就地保护。     

Genetic Diversity in Accessions of Wild Rice Oryza granulata from South and Southeast Asia

Oryza granulata, an upland wild rice species, represents an unique germplasm for possessing abilities of tolerance to shade and drought, immune to bacterial blight and resistance to brown planthopper. Although low degree of genetic variability has been revealed within its populations, little genetic information at the species level is available in determining rational conservation strategies. Here we used dominant DNA marker random amplified polymorphism DNA (RAPD) to assess the genetic variability among 23 accessions of O. granulata that collected from main distribution areas worldwide. Twenty decamer primers generated a total of 243 bands, with 83.5% of them (203 bands) being polymorphic. Calculation of Shannon index of diversity revealed an average value of 0.42 ± 0.25, indicating that O. granulata maintains a relatively high degree of genetic diversity on the species level. Analysis of genetic dissimilarity (GD) showed that genetic differentiation occurred among studied accessions, which supports the feasibility of current ex situ conservation strategies. We also suggested that information based on population studies, which could be achieved by international co-operation, is needed to conserve this widespread germplasm more effectively.     

Levels and distribution of allozyme and RAPD variation in populations of Elymus fibrosus (Schrenk) Tzvel. (Poaceae)

To study the magnitude and nature of genetic variation in E. fibrosus, the levels and distribution of allozyme and RAPD variations were investigated in populations collected from Finland and Russia. The results obtained from the allozyme and RAPD studies were compared to each other in 10 of the populations. The allozyme analysis showed that 6 of 12 presumed loci (50%) were polymorphic within the species, while the mean number of polymorphic loci within populations was 4.8%. The mean number of allele per locus for the species was 1.5 and 1.05 across the populations. Genetic diversity at the species level was low (H _es = 0.025), and the mean population genetic diversity was even lower (H _ep = 0.007). Both these values were much lower than the average for other Elymus and self-fertilising species. The largest proportion of the total allozyme diversity was found among, rather than within the populations (G _ST = 0.70). The allozyme genetic distances between the populations did not reflect geographic distances. Cluster and principal coordinates analyses revealed the same allozyme relationship patterns among the populations. A comparison of allozyme and RAPD variation in 10 of the populations showed differences in the amount of genetic variation. The RAPD analysis revealed higher levels of variation (A _p = 1.19, P _p = 20.3 and H _ep = 0.09) than the allozyme one) A _p = 1.06, P _p = 5.8 and H _ep = 0.008). For both markers, the largest proportion of the total gene diversity was found among the populations studied (G _st = 0.63 for RAPDs and G _st = 0.65 for allozyme). In contrast to the allozyme analysis, the RAPD based genetic distances did reflect geographic distances. The cluster and principal coordinates analyses showed different grouping of populations for each data set. There was a positive, but not significant, correlation (r = 0.41) between the genetic distance matrices resulting from these markers. Regional comparison revealed that the Finnish populations had a higher diversity than the Russian ones. Generally, this study indicates that E. fibrosus contains low genetic variation in its populations. The results are discussed in the context of conservation of the species.     

The Vigna angularis complex: Genetic variation and relationships revealed by RAPD analysis, and their implications for in situ conservation and domestication

The present study, using RAPD analysis, was undertaken to characterize genetic variation in three forms of V. angularis, cultivated, wild and weedy forms, and their relationships. The materials used consisted of 171 individuals (plants) or cultivars from 23 populations including 5 wild populations, 6 weedy populations, 6 cultivated populations and 6 populations with plants having wild and weedy or intermediate morphology, denoted here as complex populations. The materials used were collected on Honshu Island, Japan and seeds collected directly from the field were germinated for DNA extraction. In addition, 6 landrace accessions of V. angularis from the genebank were also analyzed. Genetic variation was highest in the wild form (H_g= 0.132; GD = 0.388), followed by the weedy form (H_g= 0.124; GD = 0.341) and the least in the cultivated form (H_g= 0.079; GD = 0.274). Intra-population genetic variation was high in the weedy and in the wild populations. However, inter-population was greater than intra-population genetic variation for all groups of populations studied in the V. angularis complex. 93% of the total diversity in the present study was exhibited by plants from complex populations and specific RAPD bands were found in these populations. Our results provide evidence that complex populations would be a logical focus for efforts to conserve the V. angularis complex in situ. Our results suggest that weedy populations are usually an ecotype of the wild form adapted to a different habitat.     

Genetic structure among earthworms (Lumbricus terrestris L.) from different sampling sites in western Germany based on random amplified polymorphic DNA

Earthworms are being used as bio-indicators to assess terrestrial pollution. However, it is often not known whether their populations possess a uniform genetic structure, which would allow comparison of residues or biological properties of earthworms from different sampling locations. In order to investigate this point, random amplified polymorphic DNA (RAPD) variation was surveyed in earthworms (Lumbricus terrestris) from five different sampling sites in Germany. Forty oligonucleotide RAPD primers (10 base pairs in length) were screened, three of which produced high polymorphic band patterns. A total of 61 DNA fragments were detected in 90 individuals of L. terrestris from five sampling sites with 49 (80.3%) RAPD markers being polymorphic. The genetic similarities within (band sharing rates between 0.756 and 0.795) and among the L. terrestris populations (0.635) were similar even at widely separate locations. Inter-population variation in the RAPD pattern for all five earthworm populations accounted for 37.9% of the total variation, while intra-population variation for three adjacent Saarland populations accounted for only 18.0% of the total variation. Principal component analysis (PCA) and the genetic distances of the populations confirm these results. Twenty-four percent of the genetic distance is caused by geographical isolation as shown by a test for isolation by distance. These results show that L. terrestris fulfils the genetic qualifications for a bio-indicator particularly at closely located sampling sites. However, the results also suggest that earthworm studies of widely separated locations should include genetic characterisation of the earthworm samples.     

Genetic diversity in Sorghum bicolor(L.) Moench accessions from different ecogeographical regions in Malawi assessed with RAPDs

Genetic variation within and among several Sorghum populations from different agroecological zones in Malawi were investigated using random amplified polymorphic markers (RAPDs). DNA samples from individual plants were analyzed using 35 oligonucleotides of random sequence. Twenty five of these primers allowed amplifications of random polymorphic (RAPD) loci. Overall, 52% of the scored loci were polymorphic. Every accession was genetically distinct. The analysis of molecular variance revealed that the within-region (among accessions) variations accounted for 96.43% of the total molecular variance. Observed variations in allelic frequency was not related to agroecological differences. The degree of band sharing was used to evaluate genetic distance between accessions and to construct a phylogenetic tree. Further analysis revealed that the sorghum accessions analyzed were genetically close despite considerable phenotypic diversity within and among them. It is suggested that all the sorghum landraces currently available in Malawi should be conserved both ex situ and in situ to maintain the current level of genetic diversity.