we propose a novel technical framework for investigating the biodiversity of soil-derived Streptomyces across extensive geographical scales. This study employs a combined approach of independent and dependent cultivation to capture Streptomyces and analyze its biodiversity. Streptomyces diversity of 19 soil samples collected all over mainland China was surveyed by using a cost-effective laboratory amplicon approach. Meanwhile, culture-dependent isolation methods to obtain 136 Streptomyces isolates from these soil samples and another lab-developed genomic library preparation strategy were used for sequencing their whole genomes. The isolated Streptomyces were subjected to species identification, pan-genomic analysis, exploration of biosynthetic gene clusters, and population genetic analysis. This integrated approach offers an efficient and comprehensive characterization of soil-derived Streptomyces biodiversity, which establishes the groundwork for a broader exploration of their biodiversity and evolution, further facilitating gene cluster exploration and drug development.