Acetosyringone （AS） has great influences on Agrobacterium-mediated genetic transformation. In order to improve the transformation frequency of waxy maize, in this study, shoot tips and immature embryo-derived callus of Lainongnuo 38 were collected as receptor materials, to compare the effects of pH in suspension medium, addition modes and concentrations of acetosyringone on genetic transformation of waxy maize. Results showed that adding 2 μl of 150 μmol/L AS in the wound with suspension medimn at pH 5.2 can significantly improve the transformation frequency of shoot tips of waxy maize; in genetic transformation of maize callus, adding 5 mg/L AS in infection solution can significantly improve the percentage of resistant callus.
The development of next generation sequencing(NGS)and high throughput genotyping are important techniques for the QTL mapping and genetic analysis of different crops.High-resolution melting(HRM)is an emerging technology used for detecting single-nucleotide polymorphisms(SNPs)in various species.However,its use is still limited in maize.The HRM analysis was integrated with SNPs to identify three types of populations(NIL population,RIL population and natural population),and the useful tags were screened.The patterns of temperatureshifted melting curves were investigated from the HRM analysis,and compared these with the kit.Among all 48 pairs of primers,10 pairs of them were selected:six pairs of primers for the NIL population,three pairs of primers for the RIL population,and one pair of primer for the natural population.The marker for the natural population was developed with a matching rate of 80%for the plant height trait,based on the data of the phenotypic characteristics measured in the field.This study provides an effective method for maize genotyping in the classification of maize germplasm resources,which can be applied to other plants for high-throughput SNP genotyping or further mapping.
