Specimens of 110 unrelated individuals were collected from Han ethnic group in Jiangsu Province of China. DNA was extracted from blood specimens using Chelex 100 method. (1) Amplifications were carried out by using reagents provided in DNA examining Kit (Codon Biotechnology CD., LTD, China) and using a Mastercycler gradiend 5331 (Eppendorf AG, Hamburg, Germany) according to the technical manual. The amplified products were separated denaturing polyacrylamide gel electrophoresis with continuous buffer system and visualized by silver staining. Data were analyzed using POWERSTATS software (2). A modified 2-test was used to verify whether the genetype distribution conformed to Hardy-Weinberg equilibrium predictions (3).
Wheat end-use product quality is highly influenced by the composition and quantity of high- and low-molecular-weight glutenin subunits (HMW-GS and LMW-GS). In the present study, 224 Chinese wheat cultivars and advanced lines were characterised for the HMW-GS and LMW-GS with allele-specific PCR markers and sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that 56 cultivars (25.0%) carried the allele Glu-D1-1d (Dx5), while 80 cultivars (35.7%) with the allele Glu-B1-2a (By8) produced a 527-bp specific band. Fourteen genotypes (6.3%) with the allele Glu-B1e (Bx20) yielded a 701-bp amplicon with the marker Mar and a 753-bp specific PCR fragment with the marker ZSBy9aF1/R3. Glu-B1h (Bx14+By15) was present in only 1 genotype, and 2 cultivars contained the allele Glu-B1f (Bx13+By16) identified with the marker ZSBy9F2/R2. Four genotypes (1.8%) with the allele Glu-B1-1d (Bx6) gave 695-bp and 830-bp bands, and 5 genotypes (2.2%) with the allele Glu-B1i (Bx17+By18) amplified a 659-bp fragment using the marker Bx. One hundred and six cultivars (47.3%) had the allele Glu-B1-2b (By9), amplifying a 663-bp fragment with the marker ZSBy9aF1/R3; 34 genotypes (15.8%) contained the allele Glu-B3d, generating a 662-bp PCR fragment with the marker gluB3d. Fifteen cultivars (7.0%) with the allele Glu-B3b yielded 1570-bp and 750-bp PCR amplicons with the markers gluB3b and gluB3bef, respectively. The allele Glu-B3h was found in 7 cultivars, generating a 1022-bp PCR fragment with the marker gluB3h. The genotypes detected by SDS-PAGE were mostly consistent with those identified by molecular markers, confirming the utility of the molecular markers. The information for the HMW-GS and LMW-GS in Chinese wheat cultivars will be useful in wheat breeding programs.
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