Genetic diversity of anchote (Coccinia abyssinica (Lam.) Cogn.) from Ethiopia as revealed by ISSR markers

Anchote (Coccinia abyssinica) is plant endemic in Ethiopia with a high calcium content grown for its edible tuberous roots. In spite of its importance as food security crop, there is no information available on molecular genetic diversity of this crop. Therefore, the aim of this study was to assess genetic diversity within and among 12 populations of anchote using ISSR markers. Using nine ISSR primers, a total of 87 scorable bands was generated of which 74 were polymorphic. Within population diversity based on polymorphic bands ranged from 13.8 to 43.53 % with a mean of 33.05 %, Nei’s genetic diversity of 0.04–0.156 with a mean of 0.12, Shannon information index of 0.07–0.23 with a mean of 0.175 and analysis of molecular variation (AMOVA) of 51.4 % were detected. With all diversity parameters, the highest diversity was obtained from Gimbi, Bedele and Ale populations, whilst the lowest was from Manna. AMOVA showed a 48.56 % between populations variability and significantly lower than that of within population variation. Population differentiation with F_ST was 48.56 %. From Jaccard’s pairwise similarity coefficient, Decha and Nedjo were most related populations exhibiting 0.76 similarity and Manna and Nedjo were the most distantly related populations with similarity of 0.52. The only pentanucliotide primer used in the study, Primer 880 (GGAGA)3, showed a unique band in some individuals that appeared to be associated with morphological quantitative traits (lowest seed number, high protein content, largest fruit size and smallest vine length). Illubabor and Gimbi populations exhibited highest genetic diversity so that the populations should be considered as the primary sites in designing conservation areas for this crop.     

A measure of genetic diversity of goldenseal (Hydrastis canadensis L.) by RAPD analysis

Goldenseal (Hydrastis canadensis L.) is a medicinal plant valued for the treatment of sore eyes and mouths. Although cultivation of the plant has helped meet growing demand, goldenseal is still considered a threatened or endangered species throughout much of its range in North America. In an effort to assess possible conservation strategies for goldenseal genetic resources, levels of genetic diversity within and among cultivated and wild populations were quantified. RAPD analysis was used to examine six cultivated and 11 wild populations sampled from North Carolina, Ohio, Pennsylvania, and West Virginia. The average percentage of polymorphic bands in cultivated and wild populations was low (16.8 and 15.5 %, respectively), and geographic range did not predict the level of genetic diversity. Most of the genetic variation (81.2 %) was within populations; only 3.6 % was partitioned between cultivated and wild populations. Our results differed from a previous study which concluded that genetic differences were greater among than within populations. The results of the current study indicate that, although goldenseal grows clonally and in dense patches, a mixed mating system in which both selfing and outcrossing occur is also operating. We therefore suggest that the ex situ conservation of individual plants within populations, chosen carefully to account for clonal propagation in situ, is an appropriate strategy for sustaining the genetic diversity of goldenseal.     

Assessment of genetic diversity in Solanum trilobatum L., an important medicinal plant from South India using RAPD and ISSR markers

Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.     

Genetic variation in the endangered Rutaceae species <Emphasis Type="Italic">Citrus hongheensis</Emphasis> based on ISSR fingerprinting

Citrus hongheensis is a critically endangered species endemic to the Honghe river region in southeastern Yunnan, China. Its genetic diversity and differentiation were investigated using Inter-Simple Sequence Repeat (ISSR) markers. One hundred primers were screened, and a total of 245 loci were amplified from seven natural populations by 13 informative and reliable primers. Of these 245 ISSR loci, 233 were polymorphic and the detected variations revealed a relatively high level of intraspecific genetic diversity. At the population level, the mean percentage of polymorphic loci (PPB) was 36.50%, while the average expected heterozygosity (He) and Shannon diversity index (Ho) were 0.1327 and 0.1972, respectively. At the species level (across all populations), PPB was 95.10%, while He and Ho were 0.3520 and 0.5195, respectively. A high Gst value (0.6247) indicated that there is significant differentiation among populations, which was confirmed by AMOVA analysis (Φst = 0.6420). Pairwise genetic identity (I) values among populations ranged from 0.6341 to 0.7675, with a mean of 0.7008. We propose that the high level of genetic differentiation may be the result of habitat fragmentation and limited gene flow (Nm = 0.1502). For effective in situ conservation and population restoration of C. hongheensis it will be important to maintain historical processes, including high outbreeding rates, sufficient gene flow, and large effective population sizes.     

Genetic diversity analysis of yams (Dioscorea spp.) cultivated in China using ISSR and SRAP markers

Yam (Dioscorea spp.) is widely cultivated in China and many landraces are maintained by local farmers. However, there is little information available about their diversity and species identity. In this study, inter simple sequence repeat (ISSR) and sequence related amplified polymorphism (SRAP) techniques were used to assess genetic diversity within 21 yam landraces from seven cultivated populations. We observed high level of polymorphism among these landraces, specifically, 95.3 % for ISSR and 93.5 % for SRAP. Analysis of molecular variance revealed a significantly greater variation among the four yam species (40.39 %) and their populations (35.78 %) than within the populations (23.83 %). The unweighted pair group method arithmetic averages clusters and principal component analysis for 21 landraces formed four well-separated groups containing landraces of each of the four species, namely, Dioscorea opposita Thunb., Dioscorea alata L., Dioscorea persimilis Prain et Burkill, and Dioscorea fordii Prain et Burkill. The ISSR and SRAP primers were highly discriminatory among the 21 landraces; all 21 landraces could be easily differentiated using these primers. The average mean of gene flow (Nm = 0.1081) estimated from high genetic differentiation (Gst = 0.8222) suggested that gene flow among the populations was relatively restricted. The lack of genetic diversity within individual yam species suggests that it is critical to develop long-term strategies for enhancing genetic diversity within various yam species.     

Genetic diversity of the African wild rice (Oryza longistaminata Chev. et Roehr) from Ethiopia as revealed by SSR markers

Data from microsatellite markers have been extensively used for both in situ and ex situ conservation strategies by determining the level of genetic diversity of natural populations that can widen the gene pool of cultivated plants. Such conservation practices are based on understanding of the between and within population genetic variations and partitioning populations on the basis of geographic origin. Therefore, the objective of this study was to assess the genetic diversity of Oryza longistaminata Chev. et Roehr and how this variation is partitioned within and between the eight O. longistaminata populations found in the different geographic regions of Ethiopia using simple sequence repeat markers. Five microsatellite markers in 320 samples generated 64 alleles that revealed the presence of large amount of genetic variability (Ho = 0.225; He = 0.768; Na = 7.375; Ne = 6.565 and P = 0.744). The F-statistics detected by the microsatellite loci showed Fst = 0.064 and Fis = 0.743 and there was no population in Hardy–Weinberg equilibrium. The genetic diversity results obtained from this data indicated that there are high levels of genetic diversity in the populations of O. longistaminata studied and it is higher within than between populations. Among the eight populations sampled, five populations were identified as priorities for conservation strategies. Thus, national collection and conservation strategies need to consider these populations.     

Analysis of genetic diversity of Ruthenia Medic (Medicago ruthenica (L.) Trautv.) in Inner Mongolia using ISSR and SSR markers

Ruthenia Medic is tolerant to drought, cold, high salinity, resistance to trampling and high quality features. Inter-simple sequence repeat (ISSR) and simple sequence repeat (SSR) molecular markers were employed for the first time to access the genetic diversity and relationships of 30 wild Ruthenia Medic accessions obtained from Inner Mongolia in the present study. A total of 94 bands were amplified by ten ISSR primers, of which 83 (88.5 %) were polymorphic, and 57 polymorphic bands (80.4 %) were observed in 69 bands amplified by ten SSR primers. Shannon’s information index (I = 0.487), and average expected heterozygosis (He = 0.329) generated by ISSR primer were higher than that of SSR analysis (I = 0.372, He = 0.231). The study indicated that ISSR were more effective than SSR markers for assessing the degree of genetic variation of Ruthenia Medic. UPGMA cluster analysis revealed inconsistencies in the clustering patterns, as the Mantel’s test between the dendrograms for ISSR and SSR data indicated a poor fit for the ISSR and SSR data types (r = 0.0970). Whereas the pattern of clustering of the genotypes remained relatively the same in ISSR and combined data of ISSR and SSR. The results of principal components analysis also supports their UPGMA clustering. These results have an important implication for Ruthenia Medic germplasm characterization, improvement, and conservation.     

Population genetic structure of in situ wild Sorghum bicolor in its Ethiopian center of origin based on SSR markers

In situ population studies of wild relatives of crops are crucial for the conservation of plant genetic resources, especially in regions with high genetic diversity and a risk of local extinction. Ethiopia is the center of origin for sorghum, yet little is known about the genetic structure of extant wild populations. Using 9 Simple Sequence Repeat loci, we characterized 19 wild populations from five regions, 8 local cultivar populations from three regions, and 10 wild sorghum accessions from several African countries. To our knowledge, this is the most comprehensive study to date of in situ wild sorghum populations in Africa. Genetic diversity corrected for sample size was significantly greater in the wild populations in situ than in local cultivars or the accessions. Approximately 41 % of the genetic variation in the wild plants was partitioned among populations, indicating a high degree of differentiation and potential value for germplasm conservation, and the average number of migrants (N_m) per generation was 0.43. Cluster analyses showed that some wild populations were grouped by geographic region, whereas others were not, presumably due to long-distance seed movement. Four wild populations from disjunct regions formed a unique cluster with an Ethiopian accession of subsp. drummondii and probably represent a weedy race. STRUCTURE and other analyses detected evidence for crop-wild hybridization, consistent with previous molecular marker studies in Kenya, Mali, and Cameroon. In summary, in situ wild sorghum populations in Ethiopia harbor substantial genetic diversity and differentiation, despite their close proximity to conspecific cultivars in this crop/wild/weedy complex.     

The Frankincense tree (<Emphasis Type="Italic">Boswellia sacra</Emphasis>, Burseraceae) from Oman: ITS and ISSR analyses of genetic diversity and implications for conservation

DNA sequences from the ITS region of the nuclear genome and Inter-Simple Sequence Repeat markers (ISSR) were used to estimate genetic diversity among and within populations of the Frankincense tree Boswellia sacra from Dhofar, Oman. This is a culturally and ecologically relevant species that is showing symptoms of decline due to anthropic factors and, possibly, global warming. ITS sequences were 511 bp long and showed low (6.4%) variation among geographically different populations. The four selected ISSR primers yielded 93 reproducible bands, of which 91 (97.9%) were polymorphic in the 97 individual profiles obtained. Total genetic diversity (H _T) and average heterozygosity within populations (H _S) resulted fairly low (0.22 and 0.136, respectively). The accession from Wadi Dowkah, an UNESCO world heritage site, showed the lowest level of genetic diversity (H _E = 0.107), while the eastern populations from the Hasik area harboured a slightly greater amount of variation. Analysis of Molecular Variance showed that differentiation among populations was relatively high (38.1%), possibly due to the reduced gene flow between the largely isolated stands of Boswellia (N _m = 0.39). Genetic distances and AMOVA suggested a clear differentiation between the eastern and western coastal populations, while those from the internal area did not form a consistent group. For conservation, the eastern sites should be given priority as core populations harbouring significant amounts of allelic diversity. Reasons for reinforcing the more depauperated stands, such as Wadi Dowkah, with local plant material only or, alternatively, with the introduction of germplasm from genetically distinct stands are discussed.     

Genetic variability of wild apricot (Prunus armeniaca L.) populations in the Ili Valley as revealed by ISSR markers

The genetic variability of 14 wild Prunus armeniaca populations was investigated using morphological analysis and inter-simple sequence repeat markers. 10 morphological characters revealed a high level variation, especially Fruit number, Fruit weight, Seed weight and Tree height. Totally, 15 selected primers generated 155 loci, with an average of 10.3 bands per primer. Nei’s gene diversity (H _e ) and Shannon’s index of diversity (I) were fairly high at the species level (H _e  = 0.2741, I = 0.4220). High molecular and morphological variability indicated that wild apricots in the Ili Valley still maintained a relatively high level of diversity. The G _ST of 0.2275 revealed a low level of genetic differentiation among populations, and genetic variation mainly resided within populations (81.51 %), which was identified with the moderate gene flow value (N _m  = 1.6974). The relatively high intraspecific genetic diversity and low inter-population genetic differentiation was largely attributed to long-distance dispersal of pollen, continuous distribution of populations and the self-incompatible breeding system.     

RAPD-based assessment of genetic relationships among and within American ginseng (Panax quinquefolius L.) populations and their implications for a future conservation strategy

American ginseng (Panax quinquefolius) is a native North American medicinal plant that is becoming increasingly vulnerable despite government harvest restrictions. To better understand the genetic diversity and gene flow of American ginseng, we studied RAPD variation in cultivated and wild populations. Classical and Bayesian analogues of genetic diversity statistics were estimated in seven wild and two cultivated populations. The wild populations were more highly structured (G _stβ  = 0.41) than the cultivated populations (G _stβ  = 0.24). The genetic diversity within populations ranged from H _eβ  = 0.05 to 0.38. Based on genetic pairwise distances, six of the wild populations clustered with the locally-derived cultivated population, while one wild population was more similar to the non-local cultivated population than the local populations. This wild population was highly diverse (P = 1.0; U = 1.0) suggesting that it was supplemented from exotic seed. A set of eight RAPD markers was identified that differentiated plants of local and non-local origin. As a conservation strategy, we recommend that regional gene banks be established based on molecular and geographic diversity to preserve the locally adapted germplasm. These regional gene banks would serve as a conservation tool and also provide a source of genes for genetic improvement of cultivated ginseng.     

Evaluation of genetic diversity in a Camelina sativa (L.) Crantz collection using microsatellite markers and biochemical traits

A genomic DNA library enriched with GA/TC repeats from Camelina sativa variety Calena has been analysed. After sequencing of about 200 randomly selected clones, approximately 60 % of them showed to contain simple or compound microsatellites with a high number of repeats. Among all microsatellite markers analysed 15 primer pairs amplified polymorphic fragments. Forty C. sativa accessions of different origin were genotyped with 15 microsatellite markers that generated 134 alleles with an average of 8.93 alleles per locus. The observed heterozygosity (Ho) among the accessions ranged from 0.0 to 0.15 with an average of 0.0370, whereas the average of expected heterozygosity (He) among accessions was 0.2769. The analysis of the average total heterozygosity (H_T = 0.651), the intrapopulation genetic diversity (H_S = 0.260), the interpopulation genetic diversity (D_ST = 0.391) and the coefficient of genetic differentiation among populations (G_ST = 0.574) demonstrated that 57.4 % of the genetic diversity is among the accessions, while 42.6 % resides within them. Phylogenetic tree of the 40 C. sativa accessions was constructed based on Nei’s genetic distance. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram shows, except for CAM108 and CAM170, a clear discrimination among C. sativa accessions grouping them in five subgroups. ANOVA analysis indicates significant differences in some biochemical and agronomic parameters among the C. sativa accessions grouped according to Nei’s genetic distance. The result of the Tukey HSD test demonstrated that the A4 subgroup showed a significant higher TWS and linoleic acid (LA) content, while the subgroup A1 showed a significant higher linolenic and lower LA content compared to the remaining groups.     

Genetic diversity and structure of natural and agronomic switchgrass (Panicum virgatum L.) populations

Panicum virgatum L. (switchgrass) is an obligate outcrossing C₄ perennial prairie grass currently being pursued for the production of lignocellulosic ethanol. Commercial production of switchgrass for bioenergy has increased substantially in the United States. Understanding the degree of native genetic diversity within and among switchgrass populations will facilitate effective germplasm improvement, conservation, and management programs. In this study, the genetic diversity and differentiation among natural and agronomic switchgrass populations were analyzed at the molecular level by using random amplified polymorphic (RAPD) DNA markers. The mean genetic diversity among populations ranged from 0.051 ± 0.136 to 0.243 ± 0.214 and the mean genetic similarity among all the switchgrass populations was 0.775. The clustering pattern of switchgrass populations grouped the individuals based on their sites of origin, with agronomic cultivars predominantly separated into distinct clusters. The grouping of individuals within and across the populations was corroborated by principal component analysis. These results are consistent with previous reports for switchgrass accessions. RAPD DNA markers were suitable for quickly estimating the genetic diversity of native and agronomic switchgrass populations, and suggest that introgression of agronomic genes into natural switchgrass populations and subsequent changes in genetic structure may be detectable.     

Genetic diversity and differentiation in Chinese sour cherry <Emphasis Type="Italic">Prunus pseudocerasus</Emphasis> Lindl., and its implications for conservation

In this study, the genetic diversity and differentiation of 10 natural Prunus pseudocerasus Lindl. populations were investigated using inter-simple sequence repeat (ISSR) markers. Totally, 18 selected primers generated 150 loci, with an average of 8.33 bands per primer. The results showed that the percentage of polymorphic bands (PPB) was pretty low at the population level (PPB = 1.13–32%), but relatively high at the species level (PPB = 84%). Besides, a high level of genetic differentiation among populations was detected based on the gene differentiation coefficient (G _ST = 0.7118) and the hierarchical analysis of molecular variance (AMOVA) (Φ _ST = 64.53%, P < 0.001), in line with the low inter-population gene flow (N _m = 0.2025). Moreover, Mantel test revealed a significant correlation between genetic and geographic distances among the populations (r = 0.5272, P < 0.005). The high level of intraspecific genetic diversity was probably related with its life history traits, while its small population size and the resultant high levels of genetic drift and inbreeding might explain the low genetic diversity within populations. The relatively high inter-population genetic differentiation was largely attributed to its small population size, habitat fragmentation, the mode of pollen and seed dispersal, and geographic isolation. Based on the present study, conservation strategies were proposed to preserve this valuable natural germplasm resource.     

Analysis of genetic diversity in the endangered tropical tree species <Emphasis Type="Italic">Hagenia abyssinica</Emphasis> using ISSR markers

Genetic diversity within and among 12 populations of the dioecious tropical tree species Hagenia abyssinica (Bruce) J.F. Gmel. in Ethiopia was examined with eight inter simple sequence repeat (ISSR) primers. A total of 104 clearly scorable bands were generated, among which 84 (81%) were polymorphic. Jaccard similarity coefficient was calculated for pairwise comparisons among all 120 individuals and ranged from 0.30 to 0.88 while average within-population similarity ranged from 0.53 to 0.66. Within-population variability was estimated as percentage polymorphic loci (ranging from 52% to 87%), Shannon’s information index (0.30–0.50) and Nei’s genetic diversity (0.21–0.35). The highest variability values were obtained for one recently planted population and for one wild population growing in an undisturbed primary forest area. Significant overall differentiation among populations was detected by both Shannon’s information index (0.26) and G _ST (0.25). Relatedness among samples was estimated with a principal coordinate analysis, and relatedness among populations was estimated with a cluster analysis (UPGMA). A Mantel test indicated a significant association between genetic and geographic distances, and an autocorrelation analysis showed significant evidence of gene flow over distances up to 30 km. This study is the first of its kind for H. abyssinica, which has decreased recently in Ethiopia and now must be regarded as an endangered species. Both within-population and between-population diversity estimates are typical of outcrossing, longlived and late successional species, suggesting that recent anthropogenic disturbances have not yet had much impact on population genetic parameters. DNA marker data can, however, be used to identify the most suitable sites for in situ conservation and for collection of material for establishment of genebanks and plant improvement programs.     

A preliminary study of the genetic diversity of Bolivian oca (<Emphasis Type="Italic">Oxalis tuberosa</Emphasis> Mol.) varieties maintained<Emphasis Type="Italic"> in situ</Emphasis> and<Emphasis Type="Italic"> ex situ</Emphasis> through the utilization of ISSR molecular markers

ISSR molecular markers have been used to investigate genetic diversity of oca (Oxalis tuberosa Mol.), an Andean neglected tuber crop species. Sampling procedure allowed a preliminary study of the genetic diversity at the intra- and intervarietal levels. Twenty tuber lots conserved in situ in the microcentre of Candelaria and ex situ in the Toralapa Centre (Bolivia) were identified. Four ISSR primers amplified a total of 25 fragments of which 17 (68%) were polymorphic. These experiments show that the structure of oca varieties is mainly based upon vernacular names with a greater differentiation among tuber lots than within them, supporting agromorphological data. ISSR technique enlightened the existence of heterogeneous varieties in oca and divergence between in situ and ex situ conservation strategies. These observations are potentially linked to the different ways of management of tubers in these two conservation systems.     

Spatially structured genetic diversity of the Amerindian yam (Dioscorea trifida L.) assessed by SSR and ISSR markers in Southern Brazil

Dioscorea trifida L. (Dioscoreaceae) is among the economically most important cultivated Amerindian yam species, whose origin and domestication are still unresolved issues. In order to estimate the genetic diversity maintained by traditional farmers in Brazil, 53 accessions of D. trifida from 11 municipalities in the states of São Paulo, Santa Catarina, Mato Grosso and Amazonas were characterized on the basis of eight Simple Sequence Repeats (SSR) and 16 Inter Simple Sequence Repeats (ISSR) markers. The level of polymorphism among the accessions was high, 95 % for SSR and 75.8 % for ISSR. The SSR marker showed higher discrimination power among accessions compared to ISSR, with D parameter values of 0.79 and 0.44, respectively. Although SSR and ISSR markers led to dendrograms with different topologies, both separated the accessions into three main groups: I—Ubatuba-SP; II—Iguape-SP and Santa Catarina; and III—Mato Grosso. The accessions from Amazonas State were classified in group II with SSR and in a separate group with ISSR. Bayesian and principal coordinate analyzes conducted with both molecular markers corroborated the classification into three main groups. Higher variation was found within groups in the AMOVA analysis for both markers (66.5 and 60.6 % for ISSR and SSR, respectively), and higher Shannon diversity index was found for group II with SSR. Significant but low correlations were found between genetic and geographic distances (r = 0.08; p = 0.0007 for SSR and r = 0.16; p = 0.0002 for ISSR). Therefore, results from both markers showed a slight spatially structured genetic diversity in D. trifida accessions maintained by small traditional farmers in Brazil.     

Assessment of genetic diversity in Amomum tsao-ko Crevost & Lemarié, an important medicine food homologous crop from Southwest China using SRAP and ISSR markers

Amomum tsao-ko Crevost & Lemarié is an important crop that has been widely used in traditional Chinese medicine and daily diets for a long time. In this study, the genetic diversity and relationships of eight cultivated populations of A. tsao-ko grown in Southwest China were examined using sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeat (ISSR) markers. The results showed that 139 (99.29%) of 140 and 185 (99.46%) of 186 bands were polymorphic by SRAP and ISSR primers amplification, respectively. The polymorphic information content of detected bands were 0.270 (SRAP) and 0.232 (ISSR), respectively. The average Nei’s gene diversity (H?=?0.217) and Shannon’s information index (I?=?0.348) at the species level generated by SRAP primer were higher than those by ISSR analysis (H?=?0.158, I?=?0.272). Genetic differentiation coefficients and molecular variance analysis (AMOVA) indicated that the genetic variance of A. tsao-ko mainly occurred within populations rather than among populations. The high genetic identity among populations was revealed by SRAP (0.937) and ISSR (0.963). Using UPGMA cluster analysis, principal coordinate analysis, and population structure analysis, the accessions were categorized into two major groups. Overall, results obtained here will be useful for A. tsao-ko germplasm characterization, conservation, and utilization.

     

Genetic diversity and sampling strategy of Scutellaria baicalensis germplasm resources based on ISSR

Destruction in wild resource and decline in genetic diversity of Scutellaria baicalensis are becoming a big problem urgently needed be dealt in China. To help efficiently utilize and conserve this medicinal plant resources, ISSR analysis was employed to reveal genetic diversity of 107 S. baicalensis accessions consisting of 82 wild and 25 cultivated germplasm resources from 6 populations in two main producing provinces in China. Only eighteen ISSR primers that resulted in accuracy and credibility, high polymorphism, well stability PCR products were selected. The maximal and minimal bands resulted from an ISSR primer are 35 and 18 respectively with a total of 480 bands were resolved in the agarose gels with an average of two alleles per locus. As indicated by the index of genetic diversity (Na = 1.96, Ne = 1.39, He = 0.25, I = 0.39), there were richness of genetic diversity in the collected samples and that the S. baicalensis germplasm had a large representativeness as primary collection. Cluster analysis by UPGMA and STRUCTURE placed the 107-germplasm resources into three linkage clusters. The samples from all the collection locations were clustered together, but the populations from one province were not clustered together. Compared to the RS and SW strategies, the LDSS proved to be more representative for core collection construct of S. baicalensis and the best sampling ratio was 10.3 % of the total germplasms. Finally, we found that the mini core collection by LDSS was composed of 11 accessions with high genetic diversity (Na = 1.82, Ne = 1.40, He = 0.25, I = 0.39). To verify the reliability of sampling strategy, the t test was used to evaluate the representativeness between core collections (constructed by LDSS), primary and reserve collection. Our results showed that no significant difference was found in the two important genetic parameters He and I (P > 0.05), thus suggesting that the LDSS will provide a rational sampling strategy for constructing a core collection of S. baicalensis germplasm resources in the future.     

Genetic diversity and relationships of ancient Chinese fir (<Emphasis Type="Italic">Cunninghamia lanceolata</Emphasis>) genotypes revealed by sequence-related amplified polymorphism markers

Some more than a century old Chinese fir genotypes with high biomass that are now rare in China were historically called “the king of Chinese fir”. The genetic diversity and relationships of those ancient Chinese fir genotypes were investigated using morphological analysis and sequence-related amplified polymorphism markers. The morphological analysis indicated that the tree volume parameter had the maximum variable coefficient value (70.2 %). In total, 18 selected primers generated 154 bands, and 150 bands were polymorphic (97.4 %). Higher values of genetic diversity parameters (h = 0.3593, I = 0.5283) were maintained at the species level, indicating that the ancient Chinese fir in China has retained a relatively high level of genetic diversity. AMOVA indicated that 73.16 % of the variation resided within provenances. The UPGMA dendrogram and genetic structure analysis identified 3 major clusters and grouped the genotypes in agreement with their geographic origins. A Mantel test revealed a significant correlation between genetic distances and geographic distances among the genotypes (r = 0.4275, p < 0.01). Overall, we recommend a combination of conservation measures including a germplasm repository and the implementation of in situ conservation.