Abstract

Info

Title:: Optimization,Validation and Primer Screening of CDDP Molecular Marker System in Soybean

Author(s):: ?LI Qiang1; 2; SU Er-hu2; GAO Ju-lin1; LI Li-jun3; XIE Min1; YU Xiao-fang1; ?1.College of Agronomy,Inner Mongolia Agricultural University,Huhhot 010019,China;2.Maize Research Institute,Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Huhhot 010031,China;3.Seeds Management Station of Inner Mongolia,Hohhot 010010,China

Keywords:: Soybean; CDDP; Orthogonal design; System validation; Primers screening

Abstract:: ?Conserved DNADerived Polymorphism(CDDP)is a novel gene targeted marker technique.L₁₆（4⁵）orthogonal experiment design were applied to optimize CDDP-PCR amplification system of soybean in five factors of Mg²⁺, Taq DNA polymerase,primer,dNTPs and template DNA.The suitable reaction system was obtained,that was 20 μL containing 2.0 mmol·L^-1 Mg²⁺,1.5 U Taq polymerase,0.375 μmol·L^-1 primer,0.3 mmol·L^-1 dNTPs,40 ng DNA template.The optimized CDDPPCR system was tested on sixteen soybeans,and the result was stable and reliable.Amplifications and genetic verification were carried out on Jiyu 75,local black soybean and 9 F₂ progenies from Jiyu 75×local black soybean.The results showed that the site in all progenies were derived from their parents and had their parents’ deletion.From the 21 primer combinations tested,thirteen were selected with clear band patterns and abundant polymorphism.The system provides the basis for evaluation of genetic diversity,construction of genetic linkage map and molecular marker assisted breeding for the soybean.