EST-SSR Marker Development and Polymorphism Analysis of Miben Pumpkin
In order to research and develop EST-SSR markers for Miben pumpkin,36.96 Gb data were obtained after RNA extraction,cDNA library construction and transcriptome sequencing,and 157 943 unigenes were de-tected after assembling and redundancy removal in this experiment.SSR detection was performed using MISA(v1.0)software,and the results showed that 59 365 SSR loci were distributed in 41 097 unigenes,the SSR distribution frequency was 37.59％,its occurrence frequency was 26.02％,and its average distribution distance was 4 672 bp.Single nucleotide,dinucleotide and trinucleotide repeats accounted for high proportion,accounting for 90.37％of the total EST-SSR,of which dinucleotide repeats were the most(30.62％),and the dominant repeat motif was AG/CT.The DNA of three pumpkin materials MB21,2202 and M10B were used as templates for polymorphism analy-sis.The amplified products of 6 among 30 pairs of primers showed polymorphism.The development and polymor-phism analysis of EST-SSR markers in Miben pumpkin would lay foundation for molecular identification,genetic map construction and genetic diversity analysis of pumpkin varieties.
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