Development and Characterization of Microsatellite Markers for the Grain Amaranths (Amaranthus spp. L.) Development and Characterization of Microsatellite Markers for the Grain Amaranths (Amaranthus spp. L.)

by Mallory, Melanie Ann

The grain amaranths (Amaranthus hypochondriacus L., A. cruentus L., and A. caudatus L.) are important pseudocereals native to the Americas that have received increased attention for their nutritional content, specifically their balance of amino acids. The objective of this project was to produce and characterize a set of highly informative, reproducible microsatellite markers for the grain amaranths. A total of 1457 clones were sequenced from three genomic libraries enriched for the microsatellite motifs AAC, AAT and AC. Of these, 353 (24%) contained unique microsatellites. An additional 29 microsatellite loci were identified among 728 BAC-end sequences of a newly developed

amaranth BAC library. Flanking primers were designed for 319 of the microsatellite loci and all were screened on a panel of diverse amaranths, including grain and weedy Amaranthus species. A total of 179 (56%) microsatellites were polymorphic across accessions from the three grain amaranths. Among these polymorphic microsatellite loci, a total of 731 alleles were identified with average of four alleles per locus. Heterozygosity values ranged from 0.14 to 0.83 with a mean value of 0.62. Thirty-seven (21%) of the markers were polymorphic between the parents of a segregating population and were shown to be inherited in a normal Mendelian fashion based on chi-squared analysis, demonstrating the utility of these markers for linkage mapping of the amaranth genome. Phylogenetic analysis using the marker data showed A. hybridus accessions in two of the three major grain amaranth clades, suggesting the polyphyletic evolution of the three cultivated species from different A. hybridus ancestors. The microsatellite markers

reported here will be useful for further evaluating the relationships among the grain amaranths and their relatives and are an ideal resource for use in marker-assisted breeding programs, germplasm analysis and varietal identification. The transferability of these markers to A. hybridus, A. powellii, and A. retroflexus as reported here suggests that the markers may be useful to other species with the genus Amaranthus, including economically important weeds, vegetable amaranths, and ornamentals.