Isolation And Characterization Of Highly Polymorphic Microsatellite Loci For Lycoris Radiata And Analysis Of Cross-species Amplification

Lycoris radiate is a perennial herb and grow from small bulbs. The plant prefers shady and moist areas and mainly distributed in the middle and lower reaches of Yangtze River in China and other countries in Southeast Asia. Their bulbs have been used in traditional Chinese medicine for a long time. There are two main medicinal components in the bulbs, lycorine and galanthamine. Galanthamine has been widely used in medicine as a strong reversible inhibitor of cholinesterase. It has been reported that galanthamine is used in the treatment of glaucoma and Alzheimer's disease. Furthermore, L. radiate has great interest in horticulture due to its scarlet flowers in autumn and green leaves all winter long. Therefore, it is an economic plant with horticultural and medicinal values.In this study, 33 novel microsatellite loci of Lycoris radiate were isolated by modified biotin-capture method using streptavidin-coated beads. Ten polymorphic microsatellite loci from the microsatellite-enriched genomic library were newly developed across 21 L. radiate individuals. The number of alleles per locus ranged from 2 to 9. The H_O and H_E ranged from 0.238 to 0.952 and from 0.455 to 0.784, respectively. Nine loci conformed to Hardy-Weinberg equilibrium expectation (HWE) and one locus (Lyra-7) significantly deviated from the HWE expectations in the sampled population after Bonferroni's correction (adjusted P value = 0.0044). Lyra-7 deviated from the HWE could be due to the presence of null allele that was confirmed by the analyses using Micro-Checker software. None of the loci showed significant linkage disequilibrium.Cross-species amplification of 10 microsatellite markers was carried out in five congeneric species, L. sprengeri, L. anhuiensis, L. albiflora, L. longituba, L. chinensis. Three to Five individuals of each species were screened at 10 loci. Five individuals of each species were screened at 10 loci. Two loci (Lyra-5 and Lyra-6) were amplified successfully in all species, two loci (Lyra-1 and Lyra-2) were amplified in four species except L. sprengeri, and nine loci were amplified in L. albiflora except Lyra-10. These results suggest that the identified microsatellite markers will be useful in genetic studies of L. radiata and other species of Lycoris.