Spring orchid (Cymbidium goeringii) is a popular flowering plant species. There have been few molecular studies of the genetic diversity and conservation genetics on this species. An assessment of the level of genetic diversity in cultivated spring orchid would facilitate development of the future germplasm conservation for cultivar improvement. In the present study, DNA markers of intersimple sequence repeats (ISSR) were identified and the ISSR fingerprinting technique was used to evaluate genetic diversity in C. goeringii cultivars. Twenty-five ISSR primers were selected to produce a total of 224 ISSR loci for evaluation of the genetic diversity. A wide genetic variation was found in the 50 tested cultivars with Nei's gene diversity (H = 0.2241) and 93.75% of polymorphic loci. Fifty cultivars were unequivocally distinguished based on ISSR fingerprinting. Cultivar-specific ISSR markers were identified in seven of 50 tested cultivars. Unweighted pair-group mean analysis (UPGMA) and principal coordinates analysis (PCA) grouped them into two clusters: one composed the cultivars mainly from Japan, and the other contained three major subclusters mainly from China. Two Chinese subclusters were generally consistent with horticultural classification, and the third Chinese subcluster contained cultivars from various horticultural groups. Our results suggest that the ISSR technique provides a powerful tool for cultivar identification and establishment of genetic relationships of cultivars in C. goeringii.
We tested cross-species amplification of 68 existing microsatellite loci in 6 species of the Sparidae family: Acanthopagrus butcheri, Sparus aurata, Pagrus auratus, Chrysophrys major, Pagellus bogaraveo, Pagellus erythrinus and one species of Bothidae, Paralichthys olivaceus. Of the 68 loci screened, sixteen were found to be polymorphic when tested in 20 individual black sea bream, Acanthopagrus schlegeli. The number of alleles per locus ranged from 2 to 9, and the observed and expected heterozygosity ranged from 0.55 to 0.95, and from 0.58 to 0.87, respectively. Our results show that cross-species amplification of known microsatellite loci in closely related species is a highly promising source of microsatellite markers for A. schlegeli.
ISSR was applied to detect the relationship between 16 Cymbidium species, and 836 bands were amplified with 15 primers, including 227 polymorphic bands. The polymorphic percentage is 27.2%. UPGMA results showed that the genetic distance were closest between C.goeringii (Rchb.f.) Rchb.f. and C. goeringii var. longibracteatum, and C.lancifolium Hook. was far away from the other 15 species. This result is quite similar to the traditional classification, indicating that the technique could supplement some information to traditional taxonomy in the molecular level.
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