Genetic diversity of natural populations of Larix principis-rupprechtii in Shanxi Province,China

Prince Rupprecht's Larch (Larix principis-rupprechtii Mayr.) is one of dominant components of middle and high elevation forests in North China. Shanxi Province is well known as “the Hometown of Prince Rupprecht's Larch” in China. In this study, six natural populations of this species across Shanxi were selected to investigate the genetic variation of the species using amplified fragment length polymorphism (AFLP) markers. Results showed that in comparison with some other species of Larix, higher genetic diversity was revealed at the species level for L. principis-rupprechtii (percentage of polymorphic loci PPL = 71.9%, Nei's gene diversity H_E = 0.225, Shannon information index I = 0.341). Most of genetic variation existed within populations (80.5%), while the genetic differentiation among populations was significant (p < 0.001) and higher (G_st = 0.194) than most other species of Larix. The differentiation can be attributed to the limited gene flow (N_m = 1.035) among populations, which could be due to the spatial isolation and habitat fragmentation. The six populations can be divided into three groups based on the Nei's genetic distances between populations (from 0.033 to 0.076). There was no significant correlation (r = 0.268, p > 0.05) between genetic distance and geographic distance among populations. The measures for in-situ or ex-situ conservation should be taken to preserve the genetic diversity of this species.     

Genetic variation and clonal diversity in populations of Nelumbo nucifera (Nelumbonaceae) in central China detected by ISSR markers

To obtain accurate estimates of population structure for purposes of conservation planning for wild lotus (Nelumbo nucifera Gaertn.) in central China, genetic diversity among and within six populations, and clonal diversity within another two populations of the species were analyzed. The genetic diversity was high (percentage of polymorphic bands, PPB = 90.0%; Shannon's information index, I = 0.383 ± 0.234) at the species level, but low within individual study populations (PPB = 35.8%; Shannon's information index I = 0.165 ± 0.241). The mean coefficient of gene differentiation (G_st) was 0.570, indicating that 43.0% of the genetic diversity resided within the population. Analysis of molecular variance (AMOVA) indicated that 50.47% of the genetic diversity among the study populations was attributed to geographical location while 12.3% was attributed to differences in their habitats. An overall value of mean estimated number of gene flow (N_m = 0.377) indicated that there was limited gene flow among the sampled populations. The level of clonal diversity found within the populations was considerably high (Simpson's diversity index, D = 0.985) indicating that clonal diversity contributes to a major extent to the overall genetic variation in the genetic structure of N. nucifera. On the basis of the high G_st and D values detected in this study we recommend that any future conservation plans for this species should be specifically designed to include those representative populations with the highest genetic variation for both in situ conservation and germplasm collection expeditions.     

Genetic diversity and structure of natural Pinus tabulaeformis populations in North China using amplified fragment length polymorphism (AFLP)

Chinese pine (Pinus tabulaeformis carr.), endemic to China, is a conifer species with extensive and fragmented distribution in North China. In this study, the genetic diversity and structure of 20 natural populations of this species were investigated using amplified fragment length polymorphism (AFLP) markers. A total of 445 fragments were revealed with 8 pairs of primers, 379 (85.17%) of which were polymorphic. A moderate level of genetic diversity was detected at the species level (Shannon's information index I = 0.356, Nei's gene diversity H_E = 0.271) and at the population level (I = 0.219, H_E = 0.206). Most of genetic variation was within populations while a considerable level of genetic differentiation was detected (G_ST = 0.352, Ф_ST = 0.304). The high differentiation could be attributed to the complex and fragmented habitats, and a limited gene flow among populations (N_m = 0.572). The Mantel test indicated that there was significant correlation (r = 0.455, P < 0.001) between Nei's genetic distance and geographical distance among all the populations. The results suggested that proper countermeasures should be taken to prevent the habitat further deterioration and maintain the genetic diversity of this species.     

Genetic diversity and structure of the critically endangered tree Dimorphandra wilsonii and of the widespread in the Brazilian Cerrado Dimorphandra mollis: Implications for conservation

We performed a comparative analysis of the genetic diversity and structure of two congeneric tree species, one critically endangered, with only 21 known individuals in the wild, Dimorphandra wilsonii, and the other widely distributed Dimorphandra mollis. Eight populations of D. mollis and all known trees of D. wilsonii, from three areas, were screened for variability with ISSR markers. Percentage of polymorphic bands, Nei's gene diversity and Shannon's index were considerably lower in D. wilsonii (P = 40.0%, h = 0.124 and I = 0.190), as compared to D. mollis (P = 70.4%, h = 0.190 and I = 0.297). Bayesian clustering showed that D. wilsonii individuals are clustered in three populations, which had high differentiation among them. Several measures for their conservation were suggested: protection of all extant populations, ex situ conservation of seeds, production of saplings in nurseries and foundation of new populations in reserve areas.     

Genetic variation of Sherardia arvensis L. – How land use and fragmentation affect an arable weed

The distribution of arable weeds extends over regions, where the species occur naturally in different kinds of habitats and regions, where they are mainly limited to arable fields.Here, we present a comparative study on the genetic structure of the arable weed Sherardia arvensis L. comprising populations from Mediterranean grasslands in Southern France and populations from arable fields in Germany. Enhanced by intensified land use since the 1960th, overall population density in Germany is very low compared to the density of populations in Southern France. We tested whether genetic variation within and among populations differ between France and Germany due to different patterns of distribution and land use. Therefore, we analysed 231 individuals of S. arvensis from 24 populations using AFLPs. Based on fragment analysis data we compared spatial genetic structure and genetic variation of populations from the two regions.Genetic variation within populations from the two regions (Shannon Index = 0.13 for both) and genetic variation among populations (26.8% and 30.0% in an analysis of molecular variance) were comparable. In both regions a drift-migration model supported the assumption of gene flow between populations. However, a clear correlation of geographical and genetic distances could only be reported for the indigenous populations from France (r = 0.46; P = 0.02), whereas in Germany a spatial genetic relationship between populations was missing (r = 0.16; P = 0.21).Our study revealed that neither French nor German populations are genetically impoverished. For French populations further the spatial genetic structure suggests that there is current gene flow between populations through pollinators and seed dispersal by cattle. For German populations comparable levels of genetic diversity and gene flow were detected, but gene flow was random. This can be traced back in all likelihood to diffuse dispersal by agriculture and the mechanical reshuffling of the individuals from the soil seed bank.     

Low genetic diversity and significant structuring in the endangered Mentha cervina populations and its implications for conservation

Eighteen populations of the endangered aromatic and medicinal plant Mentha cervina (Lamiaceae) were sampled across its natural range, in the western half of the Iberian Peninsula, and inter-simple sequence repeats (ISSRs) markers were used to assess genetic diversity and population structure. M. cervina populations exhibited a relatively low genetic diversity (percentage of polymorphic loci PPB = 14.2–58.3%, Nei's genetic diversity H_e = 0.135–0.205, Shannon's information index I = 0.08 − 0.33). However, the genetic diversity at species level was relatively high (PPB = 98.3%; H_e = 0.325; I = 0.23). The results of the analysis of molecular variance indicated very structured populations, with 50% of the variance within populations, 44% among populations and 6% between regions defined by hydrographic basins, in line with the gene differentiation coefficient (G_ST = 0.532). A Mantel test did not find significant correlation between genetic and geographic distance matrices (r = 0.064), indicating that isolation by distance is not shaping the present genetic structure. The levels and patterns of genetic diversity in M. cervina populations were assumed to result largely from a combination of evolutionary history and its unique biological traits, such as breeding system, low capacity of dispersion, small effective size and habitat fragmentation. The high genetic differentiation among populations indicates the necessity of conserving the maximum possible number of populations. The results also provide information to select sites for ex situ conservation. Optimal harvesting strategies, cultivation and tissue culture should also be developed as soon as possible to guarantee sustainable use of the species under study.     

Genetic diversity and population structure of the endangered conifer Taxus wallichiana var. mairei (Taxaceae) revealed by Simple Sequence Repeat (SSR) markers

Taxus wallichiana var. mairei is an endangered conifer with important medicinal value in southern China. Nuclear SSR markers were employed to assess genetic diversity and structure of 13 geographically disjunct populations. The present study revealed a moderate genetic diversity (H_E = 0.538) and low genetic differentiation (F_ST = 0.159). And most populations encountered in severe inbreeding and bottleneck effect. No significant genetic structure was detected by IBD and Structure analysis, which was supported by AMOVA analysis. The present results could be ascribed to an earlier period of more pronounced gene flow when the species had a more continuous distribution. However, the 13 studied populations were divided into four clusters based on the UPGMA dendrogram; these clusters were almost congruent with their geographical distributions. Vital areas such as southern mountains of Sichuan basin, Nanling Mts. and the margin of this yew's distribution range had a high priority for conservation.     

Phytophthora palmivora from Sulawesi and Java Islands,Indonesia, reveals high genotypic diversity and lack of population structure

Phytophthora palmivora is the causal agent of cocoa black pod disease, one of the primary diseases of cocoa in Indonesia. A better understanding of P. palmivora population genetics is needed to aid the development of relevant disease management strategies. This study is the first population genetic study of P. palmivora in Indonesia using microsatellite markers based on the alleles genotyping method. The microsatellite markers were used to determine the genotype of 44 P. palmivora isolates from Sulawesi (24) and Java (20) islands. The total number of observed multilocus genotypes (MLG) from both populations was 34. The genotypic diversity of P. palmivora from Sulawesi (2.90; 16.0; 0.938) and Java (2.76; 14.3; 0.930) islands was high as seen from Shannon's diversity index (H), Stoddart and Taylor's Index (G), and Simpson's Index (λ) respectively. Evenness and Nei's unbiased gene diversity exhibited similarly high levels from both populations. The linkage disequilibrium test indicated that sexual recombination occurred in the Java population (P = 0.312). Analysis of molecular variance (AMOVA) and Bayesian clustering revealed five genetic clusters, and isolates from both islands were evenly distributed across the five gene clusters. All genetic diversity was from within individuals. P. palmivora from Sulawesi and Java showed a high genotypic diversity but a lack of genetic differentiation among populations (Fst = 0.006). Both populations formed one highly diverse group. Minimum spanning network analysis showed no particular grouping of MLGs, and shared MLGs from both populations indicated long-distance migration of P. palmivora facilitated by human activities.     

Genetic diversity of the endangered Chinese endemic plant Monimopetalum chinense revealed by amplified fragment length polymorphism (AFLP)

Monimopetalum chinense Rehd. is an endangered woody vine endemic to eastern China. Using amplified fragment length polymorphism (AFLP) markers, we examined levels of genetic variation within and among eleven populations located across the species’ distribution. Although modest levels of heterozygosity were detected, other measures of genetic diversity registered relatively high levels of variability, both at the species level (P = 91.0%, H_E = 0.232, I_S = 0.365) and at the population level (P = 53.0%, H_E = 0.155, I_S = 0.239). Populations also exhibited high levels of genetic differentiation (Nei’s genetic diversity analysis, G_ST = 0.330), corresponding to isolation-by-distance and hierarchical population structure. These results indicate that, despite low levels of gene flow, populations of M. chinense still harbor substantial amounts of genetic diversity. Management plans for the species should include measures that ensure genetic diversity remains high within and among extant populations.     

Genetic diversity and structure of the noxious alien grass Praxelis clematidea in southern China

Praxelis clematidea (Asteraceae), a plant species native to South America, is a noxious weed in southern China. We examined the genetic variation and population structure of 12 populations (76 individuals) of P. clematidea from Fujian, Guangdong, and Hainan Provinces in China using inter-simple sequence repeat (ISSR) analysis. From an initial set of 69 ISSR primers, 10 were selected which yielded 80 reproducible bands. Polymorphic bands (P) were 100%, Shannon's information index (I) was 0.4226, and Nei's gene diversity (H) was 0.2791. We infer that the high levels of genetic diversity exhibited by P. clematidea may have contributed to its invasiveness. Gene flow among populations was 2.4930, which has led to homogenization. The coefficient of population differentiation (Gst = 0.1671) indicated low levels of genetic variation among populations and high levels of genetic polymorphism within populations. There was a negative correlation between population elevation and genetic diversity, while there was a significant positive correlation between genetic distance and geographic distance based on a Mantel test (r = 0.5820, P < 0.01). Some populations from different provinces clustered together in principal coordinate and UPGMA analyses indicating that human-mediated events may have contributed to the dispersal of the species.     

An observation of the loss of genetic variability in prenant’s schizothoracin,Schizothorax prenanti,inhabiting a plateau lake

Prenant’s schizothoracin (Schizothorax prenanti) was found naturally distributed in a plateau lake (Lake Wuxu) in upper reaches of the River Jiulong. The genetic diversities of the populations from L. Wuxu and R. Jiulong were investigated according to analysis of mtDNA control region and compared against three other reported river populations. The L. Wuxu population showed lower haplotype (h = 0.74) and nucleotide (π = 0.001) diversities compared to the R. Jiulong population (h = 0.87; π = 0.012); the lake population had suffered from a bottleneck, which might contribute to its low genetic diversity; the lake population significantly differentiated from the river populations, and significant gene flow was hardly found between river populations and L. Wuxu population, suggesting that there is a strong population structure between the lake and river populations. The L. Wuxu population was ecologically isolated from the R. Jiulong population, while the relatively low genetic variability had not affected its viability.     

Genetic diversity and population structure of Swertia tetraptera (Gentianaceae), an endemic species of Qinghai-Tibetan Plateau

Swertia tetraptera Maxim is an annual alpine herb endemic to the Qinghai-Tibetan Plateau (QTP). Its populations are locally scattered as isolated patches throughout this region. Genetic variation within and among thirty-four populations of this species was assessed using ISSR fingerprinting with 10 primers. High levels of genetic diversity exist within species (P = 98.9%, I = 0.3475; He = 0.2227), while the within-population diversity is low (P = 32.7%, I = 0.177; He = 0.12). High levels of genetic differentiation were detected among populations based on various statistics, including Nei’s genetic diversity analysis (G_ST = 0.4608), Bayesian analysis (θ^B = 0.476) and AMOVA (F_ST = 0.57). That is, populations shared low levels of genetic identity (I = 0.2622–0.0966). This genetic structure was probably due to severe genetic drift, breeding system and limited gene flow. The observed genetic structure of the populations implies that different populations across the distribution range of the species should be sampled to maintain high genetic diversity when a conservation strategy is implemented.     

Genetic variability of the narrow endemic Rhamnus persicifolia Moris (Rhamnaceae) and its implications for conservation

Rhamnus persicifolia Moris is an endemic small tree belonging to the Rhamnus cathartica group, growing along mountainous streams of Central-Eastern Sardinia (Italy). ISSR markers were used to detect the genetic diversity within and among six populations representative of the species distribution range. In spite of the limited distribution of this endemic taxon, fairly high levels of genetic diversity were detected. Percentage of polymorphic bands (PPB), gene diversity (H_S and H_T) and Shannon information measure (Sh) were calculated both at population (PPB = 30.70%, H_S = 0.1105, Sh = 0.1646) and at species level (PPB = 68.42%, H_T = 0.2066, Sh = 0.3139).The existence of a spatial distribution of genetic diversity in R. persicifolia was revealed by a low gene flow, a fairly high level of genetic differentiation (G_ST = 0.4583) among populations and a positive correlation between genetic and geographic distances (Mantel test, r = 0.71, p = 0.016). The spatial genetic structure was also confirmed with BAPS analysis. Our results show that a certain level of isolation by distance and sex-ratio bias may explain the distribution of genetic diversity among populations.Conservation measures are suggested on the basis of the genetic diversity detected, by implementing an integrated in situ and ex situ conservation program for each population, in order to ensure effective protection for this endemic species.     

Genetic diversity of Lilium tsingtauense in China and Korea revealed by ISSR markers and morphological characters

To study the genetic diversity and population structure of Lilium tsingtauense Gilg (Qingdao Lily), we collected 648 samples from 12 sites in China and Korea, and analyzed their Inter-Simple Sequence Repeat (ISSR) molecular markers and morphological characters. ISSR data revealed a relatively high genetic diversity at the species level, with 72.31% polymorphic loci, effective numbers of alleles of 1.437, average expected heterozygosity of 0.231 and Shannon’s information index of 0.369. Considerable genetic differentiation among the natural populations (G_ST = 0.144) and the gene flow (N_m = 1.487) were detected. AMOVA analysis and UPGMA-dendrogram suggested a hierarchical regional structure among populations, and spatial autocorrelation analysis showed a micro-scaled spatial structure. Furthermore, there was a high correlation between morphological characters and genetic parameters obtained from ISSR parameters. There was only a low genetic differentiation among the different morphological types of L. tsingtauence in China. Based on these findings, we recommend in situ and ex situ conservation strategies for the preservation of L. tsingtauense.     

Genetic variation within the sand-fixation species Caragana microphylla (Leguminosae) in Horqin sandy land detected by inter-simple sequence repeats analysis

Caragana microphylla is the most dominant and constructive shrub species in the Horqin sandy land in the northeast of China. We evaluated it's the level of genetic variation within and among populations sampled from two different populations types in Horqin sandy land by using inter-simple sequence repeat polymorphism (ISSR) molecular markers. The results showed that eight ISSR primers generated 106 bands, of which 87 were polymorphic. At the species level, genetic diversity was relatively high (P = 82.08%, h = 0.2831, I = 0.4233). Genetic variation in natural populations (h = 0.2152, I = 0.3169) was more than that in plantation populations (h = 0.2021, I = 0.3040). Based on Nei's G_ST value, more genetic differentiation among plantation populations was detected (G_ST = 0.7787). Six populations of C. microphylla clustered into two clades. These results have important implications for restoring and managing the degraded ecosystem in arid and semi-arid areas.     

The value of small habitat islands for the conservation of genetic variability in a steppe grass species

The habitat loss and fragmentation due to agricultural land-conversion affected the steppe throughout its range. In Ukraine, 95% of steppe was destroyed in the last two centuries. Remaining populations are confined to few refuges, like nature reserves, loess ravines, and kurgans (small burial mounds), the latter being often subject to destruction by archeological excavations.Stipa capillata L. is a typical grass species of Eurasian steppes and extrazonal dry grasslands, that was previously used as a model species in studies on steppe ecology. The aim of our research was to assess genetic diversity of S. capillata populations within different types of steppe refuges (loess ravines, biosphere reserve, kurgan) and to evaluate the value of the latter group for the preservation of genetic diversity in the study species.We assessed genetic diversity of 266 individuals from 15 populations (nine from kurgans, three from loess ravines and three from Askania-Nova Biosphere Reserve) with eight Universal Rice Primers (URPs).Studied populations showed high intra-population variability (I: 0.262–0.419, PPB: 52.08–82.64%). Populations from kurgans showed higher genetic differentiation (Φ_ST = 0.247) than those from loess ravines (Φ_ST = 0.120) and the biosphere reserve (Φ_ST = 0.142). Although the diversity metrics were to a small extent lower for populations from kurgans than from larger refugia we conclude that all studied populations of the species still preserve high genetic variability and are valuable for protection. To what extent this pattern holds true under continuous fragmentation in the future must be carefully monitored.     

Microsatellite Polymorphisms in Cassava Landraces from the Cerrado Biome, Mato Grosso do Sul, Brazil

Using nine microsatellite loci, we investigated genetic structure and diversity in 83 Brazilian cassava accessions, including several landraces, in the Cerrado biome in Mato Grosso do Sul, Brazil. All nine loci were polymorphic, averaging 6.00 alleles per locus. Treating each of seven municipalities as a cassava group or population, they averaged 3.5 alleles per locus, with 97% polymorphic loci, high values for observed heterozygosity (0.32) and gene diversity (0.56). Total genetic variability was high (0.668), and most of this genetic variability was concentrated within municipalities (0.577). Cluster and structure analyses divided accessions into two major clusters or populations (K = 2). Also, a significant genetic versus geographic correlation was found (r = 0.4567; P < 0.0260). Migratory routes in the Cerrado are considered main contributors to the region’s high cassava diversity and spatial genetic structure, amplifying interactions between traditional farmers and the evolutionary dynamics of this crop.     

Genetic diversity of SSR markers in wild populations of Tapiscia sinensis,an endangered tree species

Tapiscia sinensis is a Tertiary relict and endangered tree species with unique scientific research value and great economic value. In this study, we assessed the genetic diversity of five wild T. sinensis populations from different geographical regions using 10 polymorphic simple sequence repeat (SSR) markers. Our results reveal that the natural populations of T. sinensis have rich genetic diversity (PPL = 100%, He = 0.6904, I = 1.4368), with Shannon's index indicating that the T. sinensis populations are at a relatively stable stage. Of the genetic relationships among populations, the distance between the Hunan Yanling (YL) and Guizhou Xifeng (XF) populations is the smallest (0.4829); the genetic distance between the Shaanxi Ningshan (NS) and the Guizhou Xifeng (XF) populations is the largest (0.9821). A Mantel test shows that there is no correlation among the populations between geographic distance and genetic distance. AMOVA suggest that 33.3% of the genetic variation arose among the populations, while 66.7% of the variation arose within them. The moderate gene flow among populations (Nm = 0.7274) is not sufficient to counteract genetic drift within the populations and result in significant differentiation (Fst = 0.2987). Our results will benefit the conservation and exploitation of T. sinensis and provide a theoretical basis for further study of the evolution and phylogeography of the species.     

High genetic diversity in fragmented Iris pumila L. populations in Ukrainian steppe enclaves

Habitat fragmentation can prevent gene flow between plant populations and lead to a loss of genetic diversity. However, such impact of fragmentation has not always been consistently confirmed by previous studies and the issue still needs further research. Particularly little is known about the impact of fragmentation on steppe plants. Steppe once covered vast, continuous areas, and nowadays is among the most fragmented biomes. In Ukraine, remnants of this habitat survived in large but few nature reserves and loess ravines as well as on kurgans (burial mounds of ancient nomadic people), which, despite their small size, are still numerous and scattered throughout the landscape.We studied the impact of fragmentation on the genetic diversity and structure of Iris pumila, a typical species of European steppes. Our main focus was to compare the genetic characteristics between kurgan populations (n = 8), and populations from larger refugia (n = 6). We assessed the genetic diversity of the studied populations with Universal Rice Primers.Our analyses revealed high genetic diversity across all investigated populations (mean He: 0.233; mean PPB: 58.57). However in kurgan populations genetic diversity was significantly higher than in larger refugia. Genetic diversity (He) was negatively correlated with population size. Most of the molecular variance (82%) was represented within populations, whereas genetic differentiation among populations was moderate (Φ_ST = 0.160), and low among refugia types (Φ_RT = 0.026).The maintenance of high genetic diversity despite two centuries of fragmentation may be related to the moderate disturbance occurring on kurgans, which enhances the sexual reproduction of the species. Moreover, we assume that species traits such as longevity and polyploidy might counterbalance genetic drift, while its self-incompatibility and presence of a soil seed bank allows for the replenishment of the gene pool. Overall, our results suggest that kurgans can protect genetic diversity of steppe species.     

Genetic diversity and population structure of the medicinal orchid Gastrodia elata revealed by microsatellite analysis

The wild resources of Gastrodia elata are currently threatened with extinction due to over-harvesting because of their high medicinal value. Genetic diversity plays a key role in the survival of endangered orchid species. In this study we investigated the genetic pattern in eight microsatellite loci within eight G. elata populations from central China. Compared with the other orchids, G. elata showed a low level of genetic variation within populations (H_E = 0.356–0.622). The main factors responsible for the genetic pattern were the plant's inbreeding system due to mating within clone patches, and the genetic bottlenecks and genetic drift caused by a long-history over-collecting. The significant heterozygote deficit was detected in all the populations. The F statistics calculated by different approaches consistently revealed a clear genetic differentiation among populations, contributing about 20% of the total gene diversity. The results are discussed in relation to both in situ and ex situ conservation efforts of the species. The populations with a high level of genetic diversity or with great genetic distinction were identified, which should be a high priority for conservation managers.