Genetic variation in two sympatric European populations of Bosmina spp. (Cladocera) tested with RAPD markers

We used RAPDs (Random Amplified Polymorphic DNA) to test genetic divergence between two populations of Bosmina spp. in Lake Östersjön, Sweden. Previous taxonomic studies on European species within the genus Bosmina have been based on morphological characters alone. RAPD markers distinguished the two populations and supported the specific status of B. coregoni and B. longispina based on morphological characters. Furthermore, juveniles with a long antennule and a mucro were classified as B. coregoni. RAPDs also revealed genetic differences among the tested individuals, suggesting several clones within each species.     

Inter- and intraspecific genetic variation inHippophae (Elaeagnaceae) investigated by RAPD markers

Genetic diversity has been investigated by the application of molecular markers in, for the first time, all the taxa recognised in recent treatises of the genusHippophae. RAPD (random amplified polymorphic DNA) analyses were conducted with 9 decamer primers, which together yielded 219 polymorphic markers. We found 16 fixed RAPD markers, i.e. markers that either occurred in all plants of a population or were absent from all plants. Several of these markers were useful for analysis of interspecific relationships, whereas others can be considered as taxon-specific markers. Clustering of taxa and populations in our neighbour-joining based dendrogram was in good agreement with some recently suggested taxonomic treatises ofHippophae. Amount and distribution of genetic variability varied considerably between species. Partitioning of molecular variance withinH. rhamnoides supported earlier findings that a considerable part of the total variance resides among subspecies (59.6%) Within-population variability also differed considerably. Percentage polymorphic RAPD loci and Lynch and Milligan within-population gene diversity estimates showed relatively high values for some species close to the geographic centre of origin in Central Asia, e.g.H. tibetana and the putatively hybridogenousH. goniocarpa. Spatial autocorrelation analyses performed on 12 populations ofH. rhamnoides revealed positive autocorrelation of allele frequencies when geographic distances ranged from 0 to 700 km, and no or negative autocorrelation at higher distances. At distances between 700 and 1900 km, we observed deviations from the expected values with strongly negative autocorrelation of allele frequencies. A corresponding relationship between geographic and genetic distances could not be found when the analysis instead was based on one population from each of 8 species.     

Isoenzyme variation in the wild relatives ofVicia faba (Fabaceae)

Electrophoretic analysis of five enzyme systems, LAP, PGI, SKDH, SOD and 6-PGDH, among 102Vicia accessions representingV. bithynica and seven species of theV. narbonensis complex, namelyV. eristalioides, V. kalakhensis, V. johannis, V. galilaea, V. serratifolia, V. narbonensis andV. hyaeniscyamus, has been performed. The recorded variation was tentatively assigned to 41 allelic genes at eight loci; intraspecific variation was observed in all species except forV. eristalioides. The results obtained were compared with the corresponding data reported earlier forV. faba. Hierarchical grouping of the investigated taxa, includingV. faba, was based onNei's genetic identities calculated from the allozyme frequency data.Vicia faba andV. bithynica were shown to be most distantly related to one another and to the remaining species investigated.Vicia serratifolia appeared to be a peripheral member of theV. narbonensis complex. The results are discussed with reference to genetic diversity and taxonomic relationships of the species under study.     

Fruit-set and seed weight variation inAnthyllis vulneraria subsp.vulgaris (Fabaceae)

Fruit-set and seed weight variation was studied in a population ofAnthyllis vulneraria subsp.vulgaris (Fabaceae) in northwestern Spain. The plants produce several shoots, each bearing two to four inflorescences that open one at a time from bottom to top. Fruit-set, seed-pod weight and seed weight were found to be significantly higher in proximal inflorescences than in distal inflorescences of the same shoot; mean seed weight was up to three times higher in proximal than in distal inflorescences. By contrast, none of the three variables varied significantly among plants or among shoots of the same plant. Similarly, none of the three variables differed significantly between early- and late-flowering plants, or between plants monitored in 1993 and in 1994. These results are compatible with the view that shoots function as semiautonomous units as regards resource allocation, and that within the shoot resources are preferentially allocated to proximal (= early-opening) inflorescences. In the plants studied, the ratio of seed-pod weight to seed weight was fairly constant, suggesting that the pod is important for seed success.     

RAPD and morphological analysis of the rare plant species Vicia pisiformis (Fabaceae)

The amount and pattern of genetic variation was surveyed in two Swedish and three Czech populations of the rare perennial forest plant Vicia pisiformis. This species has a mainly easterly-continental European distribution and has few and small populations in Sweden. It is classified as 'vulnerable' on the Swedish Red Data list. Seeds from natural populations were collected and grown under controlled conditions in growth chambers. The variation was estimated in growth and fecundity traits and with Random Amplified Polymorphic DNA, RAPD. Low inter- and intra-population variation in RAPD-markers was found using 11 primers, with a similarity index (Wetton) for the families of 0.98. In contrast, multivariate analysis of variance showed significant morphological differences within and between populations. Also in the univariate ANOVAs, a number of the traits showed significant between-and/or within population differentiation. Cluster analysis for the morphological traits and RAPDs (UPGMA) did not structure the variation of families in accordance with their geographical distance. A Mantel test based on comparisons between Mahalonobfs and Jaccard distance for morphological and RAPD data, respectively, did not reveal any significant correlation between the two matrices. It is concluded that if a genetic conservation program is to be applied on Vicia pisiformis , different sampling strategies are needed to capture morphological vs RAPD variation. This is, to our knowledge, the first investigation that compares RAPD and morphological variation in a threatened plant species.     

Genetic variation of wild and hatchery populations of the catla Indian major carp (Catla catla Hamilton 1822: Cypriniformes, Cyprinidae) revealed by RAPD markers

Genetic variation is a key component for improving a stock through selective breeding programs. Randomly amplified polymorphic DNA (RAPD) markers were used to assess genetic variation in three wild population of the catla carp (Catla catla Hamilton 1822) in the Halda, Jamuna and Padma rivers and one hatchery population in Bangladesh. Five decamer random primers were used to amplify RAPD markers from 30 fish from each population. Thirty of the 55 scorable bands were polymorphic, indicating some degree of genetic variation in all the populations. The proportion of polymorphic loci and gene diversity values reflected a relatively higher level of genetic variation in the Halda population. Sixteen of the 30 polymorphic loci showed a significant (p < 0.05, p < 0.01, p < 0.001) departure from homogeneity and the F(ST) values in the different populations indicated some degree of genetic differentiation in the population pairs. Estimated genetic distances between populations were directly correlated with geographical distances. The unweighted pair group method with averages (UPGMA) dendrogram showed two clusters, the Halda population forming one cluster and the other populations the second cluster. Genetic variation of C. catla is a useful trait for developing a good management strategy for maintaining genetic quality of the species.     

Allozyme and RAPD analysis of the genetic diversity and geographic variation in wild populations of the American chestnut (Fagaceae)

Genetic variation among 12 populations of the American chestnut (Castanea dentata) was investigated. Population genetic parameters estimated from allozyme variation suggest that C. dentata at both the population and species level has narrow genetic diversity as compared to other species in the genus. Average expected heterozygosity was relatively low for the population collected in the Black Rock Mountain State Park, Georgia (He = 0.096 +/- 0.035), and high for the population in east central Alabama (He = 0.196 +/- 0.048). Partitioning of the genetic diversity based on 18 isozyme loci showed that ~10% of the allozyme diversity resided among populations. Cluster analysis using unweighted pair-group method using arithmetric averages of Rogers' genetic distance and principal components analysis based on allele frequencies of both isozyme and RAPD loci revealed four groups: the southernmost population, south-central Appalachian populations, north-central Appalachian populations, and northern Appalachian populations. Based on results presented in this study, a conservation strategy and several recommendations related to the backcross breeding aimed at restoring C. dentata are discussed.     

孑遗植物桫椤种群遗传变异的RAPD分析

应用 RAPD标记分析了桫椤分布在海南尖峰岭及霸王岭 ,广东塘朗山、黑石顶及大西山孑遗种群的遗传变异。 2 8个引物共检测到 1 1 8个位点 ,其中多态位点 33个 ,多态位点比率 2 7.97%。Shannon多样性、Nei遗传分化和分子方差分析 ( AMOVA)结果一致显示尖峰岭种群遗传多样性最大 ,霸王岭种群次之、塘郎山种群第三 ,黑石顶种群第四 ,而大西山种群最小。种群的遗传变异主要发生在种群间 ;Shannon指数测出的种群间遗传多样性所占比率为 79.1 0 % (基于表型频率 )和 77.85 % (基于基因频率 ) ,Nei基因分化系数 ( Gst)达 80 .69% ,分子方差分析也表明种群分化极为显著 ( Φst=0 .82 86,P<0 .0 0 1 )。桫椤的种内遗传多样性水平很低 ,种内 Shannon多样性仅为 0 .0 641 (基于表型频率 )和 0 .1 1 2 4 (基于基因频率 ) ,总基因多样性只有 0 .0 770。另外 ,Jaccard相似性系数的 UPGMA分析结果显示 5个种群分为两个分支 :尖峰岭种群和霸王岭种群组成一分支 ;黑石顶、塘郎山和大西山种群组成另一分支 ,并且三者中前两者的遗传关系更为密切。聚类分析结果还得到主成分分析的支持。根据桫椤种群的遗传变异特点 ,初步探讨了保护策略。     

Flow cytometric analysis of genome size variation in cultivated and wildPisum sativum (Fabaceae)

Fifteen cultivars, landraces, and wild accessions ofPisum sativum subspecies, and one accession ofP. abyssinicum were analysed with flow cytometry (DAPI staining) usingP. sativum Kleine Rheinländerin as internal standard. Applying the method of jointly isolating, staining, and measuring nuclei of individual seedlings of test and standard material, it was found that in allP. sativum comparisons G 1 and G 2 peaks were invariably unimodal and symmetric at coefficients of variation mostly less than 2%. This is strong evidence for absence of significant genome size variation in theP. sativum strains analysed. These data are markedly at variance to results of other authors reporting considerable genome size variation withinP. sativum. However, inP. abyssinicum flow cytograms and Feulgen densitometric measurements indicate 4–8% more DNA, at same chromosome number (2n = 14), than inP. sativum. This result demonstrates that genome size variation is indeed existent in the genus and requires further examination.     

Chromosome polymorphism,hybridization and colonization in theVicia sativa group(Fabaceae)

Vicia sativa L. s. l. is presented as a variable complex consisting of 7 subspecies, including wild types, weedy races and cultivated derivatives. The group is characterized by: (a) Decreasing aneuploid (dysploid) chromosome series (2n = 14 12 10,); (b) wide variation in karyotypes; (c) pronounced weediness and colonization of disturbed habitats; (d) cohabitation of several forms; (e) intergradations between the main morphological types; (f) predominance of self-pollination; (g) lack of full sterility between the main chromosome types. Plants with different chromosome numbers or different karyotypes are loosely interconnected by occasional hybridization, particularly in disturbed habitats. Colonization appears to be associated with reticulate evolution and the recombination of previously separated gene pools. The wide karyotypic polymorphism characteristic for this complex is interpreted as a result of such relatively recent processes.     

Comparisons of within-population genetic variation in sexual and agamospermousAmelanchier (Rosaceae) using RAPD markers

We compared genetic variation of sexualAmelanchier bartramiana and facultatively agamospermous (asexually seed-producing)A. laevis at one site where the two species are sympatric. We analyzed 77 random amplified polymorphic DNA (RAPD) markers in 29A. bartramiana individuals and 76 RAPD markers in 31A. laevis individuals. The two species do not differ significantly in mean genetic variation. However, 22.4% of genetic similarity values betweenA. laevis individuals exceed the highest value ofA. bartramiana and may represent the effect of agamospermy. Variation withinA. laevis may be the result of sexuality, hybridization, polyploidy, and other factors.     

Genome size variation inCajanus cajan (Fabaceae): A reconsideration

A recent investigation of genome size in certain samples of the pigeonpea,Cajanus cajan, indicates values from 1.55 pg to 1.99 pg (1C level), which is 1.29-fold variation between accessions. In the present analysis those of these accessions which had particularly high or low DNA contents in that study were subjected to a reanalysis using propidium iodide and DAPI flow cytometry and Feulgen densitometry. Only minor differences in genome size, not more than 1.047-fold, were found with flow cytometry, and no significant differences were obtained with Feulgen densitometry. The previously reported genome size cannot be confirmed. It is about half as large and was determined in the present study as 0.825 pg (1C, propidium iodide flow cytometry,Glycine max as standard) and 0.853 pg (1C, Feulgen densitometry,Allium cepa andPisum sativum as standards), respectively.     

中华绒螯蟹遗传变异的RAPD分析

用31个可重复性好的随机引物对中华绒螯蟹(Eriocheir sinensis)的辽河、瓯江和长江种群进行了RAPD分析。121个扩增片段中有27个多态片段。遗传距离指数（D）表明中华绒螯蟹种内遗传变异较低(D=0.004～0.063)。3个种群中,辽河种群(D=0.018)和瓯江种群(D=0.025)遗传变异较高,而长江种群遗传变异较低(D=0.013);辽河种群和瓯江种群间遗传距离(D=0.023)小于它们与长江种群间的遗传距离(D值分别为0.028和0.032)。提示人类经济活动可能已使这2个种群发生了基因交流。     

RAPD variation in relation to population size and plant fitness in the rare Gentianella germanica (Gentianaceae)

We investigated the distribution of genetic variation and the relationship between population size and genetic variation in the rare plant Gentianella germanica using RAPD (random amplified polymorphic DNA) profiles. Plants for the analysis were grown from seeds sampled from 72 parent plants in 11 G. germanica populations of different size (40-5000 fruiting individuals). In large populations, seeds were sampled from parents in two spatially distinct subpopulations comparable in area to the total area covered by small populations. Analysis of molecular variance revealed significant genetic variation among populations (P <0.001), while genetic variation among subpopulations was marginally significant (P <0.06). Average molecular variance within subpopulations in large populations did not differ significantly from whole-population values. There was a positive correlation between genetic variation and population size (P <0.01). Genetic variation was also positively correlated with the number of seeds per plant in the field (P <0.02) and the number of flowers per planted seed in a common garden experiment (P <0.051). We conclude that gene flow among natural populations is very limited and that reduced plant fitness in small populations of G. germanica most likely has genetic causes. Management should aim to increase the size of small populations to minimize further loss of genetic variation. Because a large proportion of genetic variation is among populations, even small populations are worth preserving.     

Allozyme differentiation and genetic structure of the Lady's slipper (Cypripedium calceolus) island populations in north-east Poland

The population genetic structure in the rare and endangered perennial plant, Cypripedium calceolus , from north-east Poland was investigated. A total of 11 loci were analysed, of which five were polymorphic for the species and in every population. The 11 loci resolved gave a total of 19 alleles with the mean number of 1.73 alleles per locus. The proportion of polymorphic loci (P) was the same for the species and in every population, 45.5%. The average observed and expected heterozygosities for the total Lady's slipper data set were 0.156 and 0.184, respectively. The average expected heterozygosity for the populations under study ranged from 0.176 to 0.188 and did not differ significantly among populations. Genetic diversity within C. calceolus populations from the Biebrza Valley is relatively high compared with rare taxa and taxa with similar life histories. The genetic diversity among populations of C. calceolus studied ( F _ST = 0.016) is much smaller than the genetic diversity within populations. In the species studied, there was no significant correlation between genetic and geographical distances ( r = −0.73, Mantel test). This may indicate the relatively recent origin of the populations studied, or a high level of gene flow among populations. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 433–440.     

Low levels of genetic variation inPhenakospermum guyannense (Strelitziaceae), a widespread bat-pollinated Amazonian herb

Phenakospermum guyannense is a monotypic, arborescent, long-lived monocot that is widespread in Amazonian South America. This outcrossing species is pollinated primarily by phyllostomid bats. Given these life-history characteristics,P. guyannense is expected to exhibit high levels of genetic variation and gene flow. We used isozyme electrophoresis and randomly amplified polymorphic DNA (RAPD) to characterize genetic variation in populations ofP. guyannense from French Guiana. Both measures detected a surprisingly low level of genetic variation, with only five out of twenty (25%) allozyme loci polymorphic (P), 1.35 alleles per locus (A), and an expected heterozygosity (H_e) of 0.090 at the species level. Isozymic genetic variation was even lower within populations (P = 17.5, A = 1.24, H_e = 0.074), and was corroborated by a RAPD assay that used 26 arbitrary primers (P = 3.61, A = 1.04, H_e = 0.014). Although overall levels of variation were low, the detectable variation was distributed as would be expected for an outcrossing species with extensive gene flow (mean G_ST = 0.230). We suspect thatP. guyannense is depauperate in genetic variation because of a series of bottlenecks that affected the species over this portion of its range.     

Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes

Ten snap bean (Phaseolus vulgaris) genotypes were screened for polymorphism with 400 RAPD (random amplified polymorphic DNA) primers. Polymorphic RAPDs were scored and classified into three categories based on ethidium bromide staining intensity. An average of 5.19 RAPD bands were scored per primer for the 364 primers that gave scorable amplification products. An average of 2.15 polymorphic RAPDs were detected per primer. The results show that primer screening may reduce the number of RAPD reactions required for the analysis of genetic relationships among snap-bean genotypes by over 60%. Based on the analysis of the distribution of RAPD amplification, the same number of polymorphic RAPDs were amplified from different genotypes for all RAPD band intensity levels. A comparison of RAPD band amplification frequency among genotypes for the three categories of bands classified by amplification strength revealed a measurable difference in the frequencies of RAPDs classified as faint (weakly amplifying) compared to RAPD bands classified as bold (strongly amplifying) indicating a possible scoring error due to the underscoring of faint bands. Correlation analysis showed that RAPD bands amplified by the same primer are not more closely correlated then RAPD bands amplified by different primers but are more highly correlated then expected by chance. Pairwise comparisons of RAPD bands indicate that the distribution of RAPD amplification among genotypes will be a useful criterion for establishing RAPD band identity. For the average pairwise comparison of genotypes, 50% of primers tested and 15.8% of all scored RAPDs detected polymorphism. Based on RAPD data Nei's average gene diversity at a locus was 0.158 based on all scorable RAPD bands and 0.388 if only polymorphic RAPD loci were considered. RAPD-derived 1 relationships among genotypes are reported for the ten genotypes included in this study. The data presented here demonstrate that many informative, polymorphic RAPDs can be found among snap bean cultivars. These RAPDs may be useful for the unique identification of bean varieties, the organization of bean germplasm, and applications of molecular markers to bean breeding.     

Detection of Low Genetic Variation in a Critically Endangered Chinese Pine, Pinus squamata, Using RAPD and ISSR Markers

With only 32 individuals in the northeastern corner of Yunnan Province, China, Pinus squamata is one of the most endangered conifers in the world. Using two classes of molecular markers, RAPD and ISSR, its very low genetic variation was revealed. Shannon's index of phenotypic diversity (I) was 0.030, the mean effective number of alleles per locus (A_e) was 1.032, the percentage of polymorphic loci (P) was 6.45, and the expected heterozygosity (H_e) was 0.019 at the species level based on RAPD markers. The results of ISSR were consistent with those detected by RAPD but somewhat higher (I = 0.048, A_e = 1.042, P = 12.3, H_e = 0.029). The genetic variation of the subpopulation on the southwest-facing slope was much higher than that of the subpopulation on the northeast-facing slope, which may be attributed to the more diverse environment on the southwest-facing slope. The genetic differentiation between the two subpopulations was very low. The between-subpopulation variabilities, Φ_ST, calculated from RAPD and ISSR data were 0.011 and 0.024. Because of the lack of fossil records and geological historical data, it was difficult to explain the extremely low genetic diversity of the species. We postulate that this ancient pine might have experienced strong bottlenecks during its long evolutionary history, which caused the loss of genetic variation. Genetic drift and inbreeding in post-bottlenecked small populations may be the major forces that contribute to low genetic diversity. Human activities such as logging may have accelerated the loss of genetic diversity in P. squamata.     

Isozyme variation and alleged progenitor-derivative relationships in theMedicago murex complex (Fabaceae)

Chromosomal studies ofMedicago lesinsii (n = 8) and its close relativeM. murex (n = 7) have led to the competing hypotheses that the latter is derived directly from the former, or that both originated from a common ancestor. In contrast to the relatively variableM. murex, M. lesinsii proved to be almost uniform isozymically, except that most populations of Greece differed by one allele from plants of the remainder of the range. This Greek variant ofM. lesinsii was indistinguishable from one of the isozyme variants ofM. murex. The greater level of allozyme variation inM. murex was consistent with its greater ecological amplitude and competitive ability. Also, this suggests thatM. murex is unlikely to have originated directly from the less variableM. lesinsii. The data suggest that either both species originated from a common ancestor, or that the n = 8 species evolved from the n = 7 species, a mode of chromosome evolution not previously hypothesized for the genus.     

Variability in nuclear DNA content within pigeonpea,Cajanus cajan (Fabaceae)

4C DNA values have been estimated in 16 cultivars ofCajanus cajan by cytophotometry. The values range between 6.19 pg to 7.97 pg, a 23.6% variation. The cultivars form four groups which differ significantly from each other but have insignificant difference within them. The implications of this variability with respect to the heterogeneity and origin of this legume crop are discussed.