Genetic Purity Assessment Of Hybrid Seed In Cauliflower And Waxy Maizes With Molecular Markers

Cauliflower(Brassica oleracea) and Waxy maize(Zea mays) are important vegetable crops and both waxy maize and cauliflower are cross-pollinating species which exhibited typical heterosis on quality, yield and disease resistance in F1 hybrid. RAMP, RAPD, ISSR, SSR and SRAP molecular markers were widely used in genetic breeding. In this study, we used RAMP, RAPD, ISSR, SSR and SRAP molecular markers to test the genetic purity of waxy maize and cauliflower. Our studies will provide theoretical basis and practical experience for rapidly and accurately detecting the genetic purity of hybrid seeds. The results were summarized as follows:1. Three molecular markers (RAMP, RAPD and ISSR) were used to identify seed genetic purity of cauliflower hybrid cutivar,’xinhong 65’.6 codominance primers of male and female parent-specific markers obtained. These primers(NAUSRAPem5/ pm17、NAUSRAPem8/pm17、NAURMK33/573、NAURMK33/713、NAUSSR45and NAUSSR46) were used to detect genetic purity of 210 F1 hibrid of’xinhong 65’and obtained 3 false hybrid seeds. The results showed that using SRAP、RAMP and SSR markers to identify the genetic purity of cauliflower ’xinhong 65’ was feasible.2. In this study, waxy maize hybrid ’suyunuo 2’ was used as experimental material and the SRAP, RAPD, ISSR markers were used to identify the genetic purity Genomic DNA of’suyunuo 2’hybrid cultivars and their corresponding parental lines were isolated from the young leaves with the CTAB method. We used 380 pairs of SRAP primer combination,192 RAPD random primers and 96 ISSR random primers to analyze the genomic DNA and select 5 codominance primers of male and female parent-specific markers. The five codominance primers were used to test the genetic purity of the 206 F1 hybrid cultivars of ’suyunuo 2’. The results showed that totally 4 false hybrid seeds,3 individuals only showed the FPS markers amplified and 1 individual only showed the MPS markers amplified with the 5 identified primers. These suggested that the test results using molecular marker were consistent with those test results from field, in addition to, SRAP、RAPD and ISSR markers were highly efficient and feasible for genetic purity identifition in waxy maize’suyunuo 2’ hybrid seed.3. In this study, waxy maize hybrid ’fengnuo 2146’ and their parents were used as experimental material, and three molecular markers (RAMP, RAPD and ISSR) were used to identify hybrid seed genetic purity. Two RAMP primers (NAURMN4/807 and NAURMN5/718), two RAPD primers (NAURP 706 and NAURP672) and two ISSR primers (NAUISSR880 and NAUISSR807) were identitified as codominance primers which could generate male and female parent-specific markers. The five codominance primers were used to genotype 208 F1 hybrid individuals. The results showed that four individuals only generated the FPS markers and one individual only generated the MPS markers with these five identified primers, which were in good consistency with the field identification. The results indicated that the RAMP, RAPD and ISSR could be used to identify seed genetic purity of hybrid cultivar effectively and rapidly in waxy maize.