Miscanthus sinensis has high biomass yield and contributed two of the three genomes in M.x giganteus,a bioenergy crop widely studied in Europe and North America,and thus is a potential biomass crop and an important germplasm for Miscanthus breeding.Molecular markers are essential for germplasm evaluation,genetic analyses and new cultivar development in M.sinensis.In the present study,we reported transferability of simple sequence repeat (SSR) markers from Brachypodium distachyon to M.sinensis.A set of 57 SSR markers evenly distributed across the B.distachyon genome were deliberately designed.Out of these B.distachyon SSR markers,86.0% are transferable to M.sinensis.The SSR loci amplified in M.sinensis were validated by re-sequencing the amplicons.The polymorphism information content (PIC) of the transferable SSR markers varied from 0.073 to 0.375 with a mean of 0.263,assessed based on 21 M.sinensis genotypes.Phylogenetic tree based on 162 alleles detected by 49 SSR markers could unambiguously distinguish B.distachyon from M.sinensis,and cluster 21 M.sinensis genotypes into three groups that are basically in coincidence with their geographical distribution and ecotype classifications.The markers developed by the comparative genomic approach could be useful for germplasm evaluation,genetic analysis,and marker-assisted breeding in Miscanthus.
Hua ZhaoJiangyan YuFrank M. YouMingcheng LuoJunhua Peng
DNA is one of the most basic and essential genetic materials in the field of molecular biology.To date,isolation of sufficient and good-quality DNA is still a challenge for many plant species,though various DNA extraction methods have been published.In the present paper,a recycling DNA extraction method was proposed.The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times,and correspondingly four DNA precipitations(termed as the 1st,2nd,3rd and 4th DNA sample, respectively) were conducted.This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method.This modified CTAB method was tested in eight plant species,wheat,sorghum,barley,corn,rice,Brachypodium distachyon,Miscanthus sinensis and tung tree.The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species.The DNA samples were good templates for PCR amplification of both ISSR and SSR markers.The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods,and thus is an effective and universal DNA isolation method.
