Objective: To analyze the distribution rules of SSR loci in Curcuma kwangsiensis EST

develop C. kwangsiensis EST-SSR primers

and explore the feasibility of EST-SSR for C. kwangsiensis species genetic diversity. Method: The 12 678 items of Curcuma EST (expressed sequence tag) sequences were downloaded from NCBI database

and MISA software was used to find SSR loci and select qualified sequences. Primer 5.0 software was used to design EST-SSR primers; polyacrylamide gel electrophoresis (PAGE) was used to analyze the PCR amplification characteristics of these EST-SSR primers

and further verify the rationality and validity of the developed results. Result: From 12 678 downloaded EST sequences

926 sequences contained SSR loci

accounting for 7.30% of the entire EST database. The largest proportion for SSR loci was in dinucleotide repeat sequences

followed by trinucleotide and tetranucleotide sequences

623 (50.90%)

388 (31.70%) and 125 (10.21%) respectively. A total of 165 EST-SSR primer pairs were designed according to the microsatellite sequences

and 24 of them with more than 92 points were selected for synthesis. PCR detection showed that 21 primer pairs (87.50%) could amplify stable and clear bands; in 6 different Germplasms in C. kwangsiensis

13 pairs of EST-SSR primers showed polymorphism

accounting for 54.17% in the primer design. 13 pairs of EST-SSR primers were used to verify the phylogenetic relationship for 20 C. kwangsiensis varieties. Conclusion: C. kwangsiensis EST-SSR markers have a higher development efficiency

so it is one of the important measures for development of C. kwangsiensis SSR markers

with vital significance for the identification of C. kwangsiensis variety

genetic polymorphism analysis and breeding.