Abstract: 【Objective】 Preparation of large quantity and high-quality DNA is an important prerequisite for large-scale genotypic screening and molecular marker-assisted of plant breeding. The objective of this study is to present a low-cost, high-throughput peach (Prunus persica L. Batsch) genomic DNA extraction method, meet the needs of high-throughput genetic researches and improve the working efficiency.【Method】The population were obtained from a cross between female parent ‘CN8’ (standard type, ST) and male parent ‘09-1-112’ (temperature-sensitive semi-dwarf in Prunus persica, PpTssd type) to establish a high-throughout protocol for peach DNA extraction. The F₁ segregating population were generated to assess the phenotype characteristics, resulting in observed 1﹕1 (254 standard type and 246 semi-dwarf type individuals). Subsequently, DNA extraction was carried out on the young leaves of two parents and 500 progenies by procedure using 1.2 mL thin-wall 8 strip polypropylene PCR tubes instead of a single centrifuge tube. After extraction, the quality of DNA samples was examined with ultraviolet spectrophotometry and 1% agarose gel electrophoresis, respectively. Referencing the peach genome (version 2.0) and using re-sequencing data, single nucleotide polymorphism (SNP) markers were developed and the HRM analysis was employed on F₁ population to conduct SNP genotyping. Ultimately, the extracted DNA samples were validated by using an InDel marker to verify the genotype of 500 individuals.【Result】The concentrations of DNA were in a range between 25 to 200 ng·μL^-1 and the UV absorbance ratios values (1.81-1.98) to determine DNA quality were acceptable and with high-purity. The result of agarose gel electrophoresis proved that DNA bands were clear, single with a high degree of DNA integrity. Referencing the peach genome and using whole genome re-sequencing data of two parents, SNP_Pp03_3758620 was developed in female and male parents, and the HRM analysis was employed to conduct SNP genotyping and divided temperature-sensitive semi-dwarf and standard type individuals into two groups, respectively, which proved DNA templates extracted from this DNA isolation procedure could be employed for HRM genotyping. Based on the genotype and phenotype of two parents, InDel_Pp03_3829009 was developed and the results of polyacrylamide gel electrophoresis showed that PCR amplification products showed desired fragment size, and were polymorphic in PpTssd type and ST type with bright and clear target fragment. It concluded that the extracted DNA samples could be used for indel analysis. Using this method, 1 000 samples of DNA could be extracted per day with low cost and no effect on the early growth of seedlings.【Conclusion】A simple, effective and low-cost method for extracting genomic DNA from peach was established, which can be used for molecular biology, such as genotyping, variety identification and genetic analysis. Simultaneous extraction of genomic DNA from large quantities of different samples was realized. It has high application value.

Key words: peach (Prunus persica), high-throughout, DNA extractionmethod