Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From the SSR-enriched library, 490 white colonies were randomly selected and sequenced. Among the 490 sequenced clones, 85 (17.35%) were redundant. Among the remaining 405 unique clones, 201 (49.6%) contained microsatellite sequences. We used 12 primer pairs that produced reproducible polymorphic bands for four diverse strains, and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles, and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, whereas for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among the 32 Flammulina velutipes strains on the basis of SSR data were investigated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from an SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenetic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From SSR-enriched library, 490 white colonies were randomly selected and sequenced. In the 490 sequenced clones, 85 clones (17.35%) were redundant. Among the remaining 405 unique clones, 201 clones (49.6%) contained microsatellite sequences. As a result, 12 primer pairs produced reproducible polymorphic bands within diverse 4 strains and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, while for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among 32 Flammulina velutipes strains based on SSR data were generated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

Seed & Variety Service for Plant Variety Protection (PVP). We previously constructed DNA profile database for identifying 159 lettuce varieties using 60 simple sequence repeat (SSR) markers. In this study, we selected optimum markers from previously applied markers and new SSR markers for the standardization of DNA profile database of 65 commercial lettuce varieties containing 18 PVP varieties distributed in Korea. Twenty-eight SSR markers from 60 SSR markers were selected for characterization with 65 commercial lettuce varieties according to the reproducibility, polymorphism, band pattern of marker and easiness of scoring. Out of 156 SSR primers, we additionally selected 11 new SSR primer pairs showed polymorphisms between 8 varieties and repetitive reproducibility on capillary electrophoresis system. Totally 127 polymorphic amplified fragments were obtained using 39 SSR markers. Two to seven SSR alleles were detected for each locus with an average of 3.3 alleles per locus. Average polymorphism information content was 0.517, ranging from 0.281 to 0.771. Genetic distance of clusters ranged from 0.29 to 0.96 by unweighted pair-group method with arithmetical average based on Jaccard’s distance coefficients. A total of 65 commercial lettuce varieties were discriminated by 39 SSR marker genotypes. These SSR profile database developed will be continually characterized for the standardization of DNA profiles for lettuce commercial varieties

The SSR markers were generally used to analysis the plant genetic diversity, but it have been rarely reported in case of castor bean. We constructed the microsatellite-enriched genomic library and sequenced totally 775 clones to obtain the microsatellite sequence information of the castor bean. Among the sequenced clones, one hundred fifty clones (19.3%) were redundant and four hundred twenty (67.2%) were found to contain microsatellite sequences within the remaining 625 unique clones. A total of 237 primer pairs were designed based on the sequenced microsatellite clones information and evaluated for polymorphism in ten castor bean accessions. Twenty-eight SSR markers produced reproducible polymorphic bands and were further characterized using a diverse set of 25 castor bean accessions. The majority of unique SSRs revealed dinucleotide motives (84%) on the other hand the ratio of trinucleotide motives was 15%. A microsatellite enriched library from the Ricinus communis L. was mainly consisted of [(AG), (GA)/(CT), (TC)] and [(CTT)/(AAG)] microsatellite motifs. The length of dinucleotide SSRs ranged from 4 to 50 repeats with an average 12.4, and that of trinucleotide SSRs from 4 to 56 with an average of 7.35 repeats. These newly developed microsatellite markers will be useful for breeding system and classification of Ricinus communis L.

The genus Allium which is one of the biggest plant genera comprises around 750 species that show the various morphological and ecological diversity with various taxonomic and geographical groups. The species of genus Allium has not been clearly distinguished because of extraordinary large amount of variation. We developed 8 simple sequence repeat markers (SSRs) which showed polymorphism within A. sativum accessions, but it was insufficient in transferability analysis of other Allium species. In this regard, we finally selected 50 primer pairs which was applicable to other Allium species adding to 8 primer pairs. According to results from application of 50 SSR markers to 9 Allium species, the average number of alleles ranged from 1.400 (A. porrum) to 1.889 (A. tuberrosum) and A. tuberosum (2n=32) has maximum 9 alleles. The lowest transferability value was 42.0% ( A. cepa and A. chinense) while A. sativum showed 96.7%. The A. porrum conceived to be close relationship to A. sativum as Allium subgenera revealed higher transferability (73%) rather than other Allium species. According to PCA analysis, three groups were divergent, and the A. fistulosum and A. sativum revealed the distinct groups and the rest 7 species formed another group.