High genetic diversity detected in the endemic Primula apennina Widmer (Primulaceae) using ISSR fingerprinting

Primula apennina Widmer is endemic to the North Apennines (Italy). ISSR were used to detect the genetic diversity within and among six populations representative of the species distribution range. High levels of genetic diversity were revealed both at population percentage of polymorphic band (PPB = 75.92%, H _S = 0.204, H _pop = 0.319) and at species level (PPB = 96.95%, H _T = 0.242, H _sp = 0.381). Nei gene diversity statistics (15.7%), Shannon diversity index (16.3%) and AMOVA (14%) detected a moderate level of interpopulation diversity. Principal coordinate and Bayesian analyses clustered the populations in three major groups along a geographic gradient. The correlation between genetic and geographic distances was positive (Mantel test, r = 0.232). All together, these analyses revealed a weak but significant spatial genetic structure in P. apennina, with gene flow acting as a homogenizing force that prevents a stronger differentiation of populations. Conservation measures are suggested based on the observed pattern of genetic variability.     

Conservation genetics of the endangered terrestrial orchid Liparis japonica in Northeast China based on AFLP markers

Levels of genetic diversity and population genetic structure of the rare, endangered terrestrial orchid Liparis japonica were examined for eight natural populations (n = 185) in Northeast China using six AFLP primer pairs, where this species has experienced severe habitat loss and fragmentation. Based on 406 DNA bands, a high level of genetic diversity was found at the species level with the PPB of 85.47 %, while the genetic diversity at the population level was low (PPB = 47.48 %). A significantly high degree of population differentiation was found with 42.69 % variation existed among populations as measured by AMOVA, indicating potential restricted gene flow. The genetic distances between populations were independent of the corresponding geographic distances, and the genetic relationship of individuals had no significant correlation with their spatial distribution. The restricted gene flow might be impacted by reduced population size, habitat destruction and fragmentation. The results in this study suggested that habitat protection and keeping a stable environment are critical for the conservation of L. japonica species.     

Genetic diversity of the threatened Chinese endemic plant,Sinowilsonia henryi Hemsi. (Hamamelidaceae), revealed by inter-simple sequence repeat (ISSR) markers

Sinowilsonia henryi Hemsi., the only representative of the monotypic genus Sinowilsonia Hemsi. (Hamamelidaceae), is a threatened plant endemic to China with high phylogenetical, ecological and economical values. In the present study, inter-simple sequence repeat (ISSR) markers were employed to investigate the genetic diversity and differentiation of 214 individuals sampled from 14 populations. Fifteen selected primers yielded a total of 178 bright and discernible bands. The genetic diversity was low at the population level (h = 0.1025; I = 0.1506; PPL = 26.7%), but quite high at the species level (h = 0.2449; I = 0.3690; PPL = 72.5%). In line with the limited gene flow (N_m = 0.3537), the hierarchical analysis of molecular variance (AMOVA) revealed pronounced genetic differentiation among populations (Φ_ST = 0.6639). Furthermore, the Mantel test revealed a significant correlation between genetic and geographic distances among populations (r = 0.688, P = 0.001), indicating the role of geographic isolation in shaping its present population genetic structure. The present patterns of genetic diversity of S. henryi were assumed to result largely from its evolutionary history and geographic factors. Based on these findings, conservation strategies were proposed to preserve this threatened plant.     

Genetic diversity in Artemisia halodendron (Asteraceae) based on chloroplast DNA psbA–trnH region from different hydrothermal conditions in Horqin sandy land, northern China

The genetic diversity of Artemisia halodendron (Asteraceae), a constructive and dominant species in the Horqin sandy land, was investigated to examine the genetic relationships with different hydrothermal regions in the Horqin sandy land. We sequenced chloroplast DNA (cpDNA) fragments psbA–trnH of 243 plants from ten populations across the Horqin sandy land. The analyses of cpDNA variation identified seven haplotypes. A high level of haplotype diversity (H _d = 0.831) and low level of nucleotide diversity (π = 0.0018) were detected. Haplotypes clustered into three tentative clades. Low genetic differentiation among regions was consistently indicated by hierarchical analyses of molecular variance (AMOVA).     

Fragmented habitat drives significant genetic divergence in the Chinese endemic plant,Urophysa henryi (Ranuculaceae)

Urophysa henryi (Oliv.) Ulbr., endemic to China with small populations, is known as a medicinal plant. In this study, ISSR markers were used to assess the genetic diversity and population structure throughout its entire distribution areas. Twelve primers revealed high genetic diversity at the species level (PPB = 95.6%; H = 0.3441; I = 0.5111), as well as high level of genetic differentiation (F_ST = 0.659, p < 0.001; G_ST = 0.677) and restricted gene flow (Nm = 0.239) among populations. According to the UPGMA and PCoA analysis, the 9 populations were clustered into three main groups, which were roughly in accordance with their geographical regions. In addition, a significant correlation between the genetic difference and geographic distances among populations was detected from the IBD analysis (r = 0.516, p = 0.003). These results indicated that the habitat heterogeneity and physical barriers play important roles in the modern distribution pattern and population divergence of U. henryi. However, human activities have posed serious threat to its living environment and continued survival. It is necessary to adopt some measures to restrict anthropogenic disturbances and preserve the existing populations.     

Genetic Diversity and Geographic Differentiation of the Giant Tiger Shrimp (Penaeus monodon) in Thailand Analyzed by Mitochondrial COI Sequences

Genetic diversity and geographic differentiation of the giant tiger shrimp, Penaeus monodon, in Thai waters (Satun, Trang, Phangnga, and Ranong in the Andaman Sea and Chumphon and Trat in the Gulf of Thailand) were examined by COI polymorphism (N = 128). We observed 28 COI mitotypes across all investigated individuals. The sequence divergence between pairs of mitotypes was 0.00–20.76%. A neighbor-joining tree clearly indicated lineage separation of Thai P. monodon and large nucleotide divergence between interlineage mitotypes but limited divergence between intralineage mitotypes. High genetic diversity was found (mean sequence divergence = 6.604%, haplotype diversity = 0.716–0.927, π = 2.936–8.532%). F-statistics (F _ST) and an analysis of molecular variance (AMOVA) indicated that the gene pool of Thai P. monodon was not homogeneous but genetically differentiated intraspecifically (P < 0.05). Six samples of P. monodon could be allocated into three different genetic populations: Trat (A), Chumphon (B), and the Andaman samples Satun, Trang, Phangnga, and Ranong (C). Contradictory results regarding patterns of geographic differentiation previously reported by various molecular approaches were clarified by this study.     

Genetic differentiation of Rosa rubiginosa L. in two different Argentinean ecoregions

We analyzed genetic differentiation of Rosa rubiginosa by RAPD from populations growing in two Argentinean ecoregions, Chaco Serrano and Patagonian Steppe. Leaf material was collected during the spring and summer of 2006. UPGMA dendrogram and PCoA clearly suggest a geographical differentiation of the provenances of R. rubiginosa populations. AMOVA analyses revealed high genetic variation within populations (71%) and low variation between populations (29%), in agreement with values estimated by the Shannon–Weaver index. Genetic differentiation between populations estimated by AMOVA was ?_PT = 0.29 (P < 0.001). Nei’s Gst (0.2205) was lower for interpopulation variation. The low interpopulation value obtained suggests genetic homogeneity between the populations. The presence of specific monomorphic bands accounts for the genetic differentiation between populations. The high percentage of within-population genetic diversity suggests the introduction of genetic variation into both ecoregions. From the present results we can conclude that we observed two independently established populations with high similarity between them and a strong intrapopulation differentiation. The empty niche hypothesis for explainging invasion success might explain the invasiveness of R. rubiginosa in Argentinean ecoregions.     

Genetic diversity and population structure of Butea monosperma (Lam.) Taub.- a potential medicinal legume tree

Three molecular marker systems, Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeats (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) were employed to investigate the genetic structure and diversity among the 14 natural populations of Butea monosperma collected from different geographical regions of India. Detected by 17 RAPD, 15 ISSR and 11 SRAP primer combinations, the proportions of polymorphic bands were 84.2 %, 77.2 % and 91.9 %, respectively, and the mean Nei’s genetic distances among the populations were 0.13, 0.10 and 0.13, respectively. Partitioning of genetic variability by Analysis of molecular variance (AMOVA) revealed that the high genetic diversity was distributed within the populations. AMOVA also revealed that the coefficient of gene differentiation among populations based on F_ST was very high irrespective of markers used. The overall gene flow among populations (Nm) was very low. Cophenetic correlation coefficients of Nei’s distance values and clustering pattern by Mental test were statistically significant for all three marker systems used but poor fit for ISSR data than for RAPD, SRAP and combined data set of all three markers. For all markers, a high similarity in dendrogram topologies was obtained, although some differences were observed with ISSR. The dendrogram obtained by RAPD, SRAP and combined data set of all three markers reflect relationship of most of the populations according to their geographic distribution.     

Genetic composition and differentiation of sloe (Prunus spinosa L.) populations in Germany with respect to the tracing of reproductive plant material

Sloe (Prunus spinosa L.) is a shrub native to Europe. In Germany, 50–80 % of all planted sloe is imported. Little is known about the genetic diversity patterns within and between German sloe populations. Thus, a debate arose how to avoid risks for nature and landscape by planting potentially maladapted material. The main objectives of our study are to analyse the genetic differentiation pattern of sloe populations in Germany, to identify geographic/genetic structures and to evaluate their potential for tracing reproductive material. 17 natural populations from Germany and 1 from Italy and Hungary were investigated by Amplified Fragment Length Polymorphisms (AFLP) and PCR–RFLP techniques. The AMOVA analyses based on AFLPs for all populations and for the German populations only result in equally high differentiation values of Φ_PT = 15 % of molecular variance between populations. The analysis of cpDNA PCR–RFLPs resulted in 24 haplotypes with 30 % showing genetic variation between populations. Overall values of genetic variability over all loci and populations are: Na = 0.832, Ne = 1.114 and He = 0.072. Mantel tests for AFLPs and cpDNA haplotypes reveal no association between geographic and genetic distances between populations as a result of a lack of differentiation between German populations and those from southern and southeastern Europe. Weak geographic/genetic patterns were observed on a large scale. However, these concern the German populations only. Our results indicate that vegetative regeneration in combination with founder effects may influence the level of differentiation between populations. Populations with a large amount of vegetative propagation are more differentiated from other populations than those populations which exhibit less vegetative regeneration. The assignment of reproductive material (i.e. plant material) to potential source populations resulted in high values of correct allocations. Hence, such methods can be applied to trace reproductive material of unknown origin.     

High level of genetic variation within clonal orchid Goodyera repens

Type of reproduction has an important effect on the maintenance of particular populations and species persistence in time and space. This trait significantly influences the ecological and genetic structure of populations, and in consequence the evolution of species. The primary objectives of this study were: to estimate genetic diversity within and among populations of clonal species Goodyera repens from different populations in northeastern Poland, and to amount factors shaping the genetic structure of this orchid. Based on 451 rosettes of G. repens from 11 localities in northeastern Poland, we conducted a genetic population analysis using allozymes. We included information on population size, flowering, fruit set and seed dispersal to elucidate their influences on genetic diversity of this species. Populations differed according to demographic properties. The majority of seeds (86.4–94.8 %) were found at a distance of 0.2 m. We observed a high level of genetic (P _PL = 50 %, A = 1.68, H _O = 0.210, H _E = 0.204) and genotypic diversity (G = 163, G/N _S = 0.66, G _U = 30.2 %), and low but statistically significant genetic differentiation among populations (F _ST = 0.060; P < 0.001). We suggest that the genetic diversity of G. repens is mainly an effect of the abundance of pine and spruce forest communities suitable for this species in NE Poland and the high level of sexual reproduction.     

Genetic diversity in the endangered Sicilian endemic Brassica rupestris: Proposals for a conservation strategy

Abstract

Brassica rupestris Raf. is a chasmophyte species that includes two subspecies, both endemic to Central-Western Sicily (Italy). Inter-Simple Sequence Repeat (ISSR) markers were used to detect genetic diversity within and among eight populations representative of the species' distribution range. High levels of genetic diversity were revealed both at the population (PPB = 53.88%, H _S = 0.212, Sh = 0.309) and at the species level (PPB = 96.55%, H _T = 0.307, Sh = 0.464). The correlation between genetic and geographical distances was negative (Mantel test, r = ?0.06, P < 0.95). The two subspecies of B. rupestris, subsp. rupestris and subsp. hispida, showed remarkable genetic similarity and molecular data did not unequivocally support their distinctness. The pattern of genetic variation revealed by our study bears important consequences for conservation management: It is desirable to preserve B. rupestris populations in situ with a “dynamic” strategy, while, ex situ conservation programmes might be improved to safeguard maximum genetic diversity.     

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.     

Conservation genetics in two endangered endemics from the Canary Islands, Helianthemum gonzalezferreri Marrero (Cistaceae) and Kunkeliella subsucculenta Kämmer (Santalaceae): different life histories that involve different management strategies

Helianthemum gonzalezferreri and Kunkeliella subsucculenta, two endangered endemic species from the Canary Islands, were analysed to determine the levels and structure of genetic diversity in their natural populations. The mean value of Shannon’s diversity index for H. gonzalezferreri and K. subsucculenta were I = 0.315 and I = 0.331, respectively. AMOVA and Bayesian analysis showed high genetic differentiation between populations in H. gonzalezferreri (F _ST = 0.297). This genetic differentiation was graphically shown with the principal component analysis (PCA), in which the majority of individuals were distributed into two groups. Contrarily, K. subsucculenta populations showed a considerable degree of gene exchange revealed by Bayesian cluster analysis and PCA. The genetic differentiation between the two H. gonzalezferreri populations suggests different management strategies for each population. Translocation between the two natural populations of K. subsucculenta, to increase genetic variation, would not mean outbreeding depression, since both populations share the same genetic pool.     

Assessment of genetic diversity and chemical composition among seven black locust populations from Northern China

Black locust (Robinia pseudoacacia L.) is one of the important biomass sources used to produce bioenergy and bioethanol. In this study, we examined variations in chemical composition as well as genetic diversity and differentiation among 165 black locust plants from seven populations in five provinces (Beijing, Hebei, Gansu, Shanxi, Shandong) of Northern China using microsatellite markers(SSR). The contents of cellulose, hemicellulose, lignin varied widely among seven populations. Of the microsatellite markers analysed, 14 showed polymorphisms, and 45 alleles were identified. The polymorphism information content (PIC) ranged from 0.2885 (Rops4) to 0.6837 (Rp200), and most of the microsatellite loci had PIC values > 0.5. Expected heterozygosity (He), observed heterozygosity (Ho), and Shannon's information index (I) detected relatively high genetic variation among populations. The percentage of polymorphic loci in three populations was 100%, and the average among all populations were 95.92%. Analysis of molecular variance (AMOVA) indicated that the degree of genetic differentiation among the seven populations was low (G_ST = 0.058; N_m = 4.05), and chemical compositions had no relationship with genetic or geographic distance. This study demonstrates that microsatellite markers efficiently assess of genetic diversity and genetic differentiation in black locust populations, and all seven populations exhibit high genetic diversity. The population from Feixian has the potential to be lingo-cellulosic biomass for bioenergy and bioethanol.     

Chloroplast microsatellite diversity of Opisthopappus Shih (Asteraceae) endemic to China

The genus Opisthopappus consists of two species, O. longilobus and O. taihangensis. Both of them are important economic and ornamental plants but endangered in China. In this study, genetic variation in four populations of O. longilobus and nine populations of O. taihangensis was estimated by chloroplast microsatellites (cpSSR). Low genetic diversity was detected in O. longilobus and O. taihangensis populations. The percentages of polymorphic loci (PPL), Nei’s gene diversities (h), and Shannon’s indices (I) was 53.56 %, 0.074, and 0.139, respectively, for O. longilobus, while PPL = 44.44 %, h = 0.018, I = 0.048 for O. taihangensis. Thirteen chloroplast haplotypes were identified from 153 individuals analyzed of two species. Ten haplotypes were found in O. longilobus, while seven in O. taihangensis. Haplotype 1 was the common and ancestral one. The level of haplotype diversity was also low (Hd _{O. longilobus}  = 0.4875, Hd _{O. taihangensis}  = 0.3452). AMOVA analysis revealed that the majority of the genetic differences was partitioned within populations of O. longilobus and O. taihangensis, as confirmed by PCoA and cluster analysis. Inter-population genetic distances among population of O. longilobus and O. taihangensis correlated insignificantly with geographic distances (r = 0.286, P = 0.128 for O. longilobus vs. r = 0.356, P = 0.058 for O. taihangensis). The self-compatibility, effective gene flow, narrow geographical distribution, and historical factors may be the main factors causing differentiation in the genetic structure of O. longilobus and O. taihangensis populations.     

Population Genetic Structure of the Invasive Red Swamp Crayfish in China Revealed by ITS1 Variation

The invasive red swamp crayfish (Procambarus clarkii) provides a valuable opportunity for studying the population genetics of invasive species that disperse rapidly. We analyzed the population genetic structure among 12 populations of the crayfish in China based on the internal transcribed spacer 1 (ITS1) region. The ITS1 of 815 bp aligned across 34 haplotypes; the average GC content was 53.9%. AMOVA showed that intrapopulation variation (95.26%) was much higher than interpopulation variation (4.74%). Genetic differentiation between the Taiwan and mainland populations (F _st = 0.160) was moderate, but the Chinese population (Taiwan and the mainland combined) and an American population were highly differentiated (0.682 and 0.977, respectively). Gene flow between the Chinese and American populations (N _m = 0.006 and 0.117, respectively) was lower than that between Taiwan and the mainland (1.536). Phylogenetic trees showed that three major genealogical clusters matched the sample locations well, suggesting that genetic differentiation is created largely by geographic isolation.     

Estimation of genetic variability and population structure in Sapindus trifoliatus L., using DNA fingerprinting methods

Genetic variability and population structure of Sapindus trifoliatus L. (Sapindaceae), collected from Gujarat, Karnataka and Uttar Pradesh states were estimated using three DNA fingerprinting methods viz., random amplified polymorphic DNA (RAPD), directed amplification of minisatellite DNA (DAMD) and inter-simple sequence repeats (ISSR). The cumulative data analysis carried out for all three markers showed 69.42 % polymorphism. The intra-population genetic diversity analysis revealed the highest values of Nei’s genetic diversity (0.16), Shannon information index (0.24) and polymorphic loci (43.99 %) among Bhavnagar (BH) population, whereas lowest values were found in Junagarh (JU) population. The maximum inter-population average genetic distance (0.20) was between Allahabad (AL) and JU populations. Analysis of molecular variance (AMOVA) showed highest percentage of variation among individuals of populations (56 %) followed by 25 % among populations and 19 % among regions. Principal coordinate analysis and UPGMA dendrogram revealed that genetic diversity was in congruence with the geographical diversity. The data strongly suggest that low genetic flow, geographic isolation and to some extent genetic drift are the major factors responsible for high genetic differentiation. Preservation of genetic diversity of S. trifoliatus is important, both to promote adaptability of the populations to changing environment as well as to preserve a large gene pool for future genetic improvement. The present study using RAPD, DAMD and ISSR profiles of S. trifoliatus provide the means of rapid characterization of accessions within the populations, and thus enable the selection of appropriate accessions for further utilization in conservation and prospection programs of this important plant genetic resource.     

Genetic differentiation between sun and shade habitats in populations of Lindera benzoin L.

Differences in selection patterns among habitats can alter the distribution of genetic diversity even when this is estimated with neutral markers. For plants, light is an essential resource that can influence both abiotic and biotic components of habitat. We examined genetic differentiation between sun and shade habitats in Lindera benzoin L. (Spicebush), a perennial understory shrub. Genetic diversity of 127 plants from sun and shade habitats in two populations of L. benzoin was determined using 12 polymorphic microsatellite markers. We analyzed patterns of genetic diversity using analysis of molecular variance (AMOVA), and we assessed correlation between genetic and geographic distance using Mantel tests. We found (1) low levels of differentiation among populations (F _ST = 0.028), (2) little evidence of genetic structure within populations due to isolation-by-distance, and (3) some evidence of habitat-based genetic differentiation. Specifically, the AMOVA showed a small (0.5%) but significant portion of overall variation could be explained by differences between habitats. The overall low levels of differentiation we saw were likely a result of extensive gene flow in this dioecious, bird-dispersed species.     

Genetic diversity of Aegiceras corniculatum (Myrsinaceae) revealed by amplified fragment length polymorphism (AFLP)

Aegiceras corniculatum is a cryptoviviparous mangrove tree distributed in the Indo-West Pacific. The genetic structure of 13 populations of A. corniculatum from South China, Malay Peninsula, Sri Lanka, and North Australia, was assessed by amplified fragment length polymorphism (AFLP) markers. Our results showed a relatively high level of genetic variation at the species level (P = 92%, H_E = 0.294 and H_s = 0.331 ± 0.001). The value of G_ST was 0.698, suggesting significant genetic differentiation among populations. At the population level, however, genetic diversity was low (P = 24%, H_E = 0.086 and H_s = 0.127 ± 0.001). When populations were grouped according to geographic regions, i.e., South China, Malay Peninsula and Sri Lanka, it was inferred from analysis of molecular variance (AMOVA) that about half the total variation (49%) was accounted for differentiation between regions. A UPGMA dendrogram based on genetic distance also revealed five major clades corresponding to geographical regions within the distribution of A. corniculatum, although the precise relationships among the clades were not fully concordant with expected geographical delineations and need further study.     

Phylogeography of Chrysanthemum indicum L. (Compositae) in China based on trnL-F sequences

Using chloroplast DNA (cpDNA) trnL-F sequences, we studied the phylogeographic pattern and demographic history of Chrysanthemum indicum with 81 individuals from 27 populations representing the partial geographic range in China. Fifteen haplotypes were identified based on nucleotide variation. Relatively high levels of haplotype diversity (Hd = 0.783) and low levels of nucleotide diversity (Pi = 0.00521) were detected in C. indicum. An analysis of molecular variance (AMOVA) indicated that most of the genetic variation (59.9%, P < 0.001) was partitioned within populations. Genetic differentiation among populations was high (F_ST = 0.35), suggesting that there is little gene flow among populations. Nested clade analysis showed that 15 haplotypes formed twelve 1-step clades, five 2-step clades and two 3-step clades, with four clades showing significant geographical association. We concluded that contiguous range expansion and restricted gene flow/dispersal but with some long distance dispersal seemed to have shaped the present-day distribution of the haplotypes of C. indicum in China.