This study proposes a strategy for selecting materials for Abies nephrolepis, a key subalpine species, to restore damaged forests in Mt. Gariwangsan while considering genetic diversity. The genetic diversity distribution of A. nephrolepis in Korea was evaluated using nuclear simple-sequence repeat (nSSR) markers and the genetic differences between populations were analyzed. A suitable genetic restoration population was selected and a strategy for selecting restoration materials was presented. The genetic diversity indices of 326 individuals across 10 populations, and the genetic relationships between populations, were analyzed using AMOVA, principal coordinate analysis, UPGMA, and STRUCTURE analysis. The genetic diversity of A. nephrolepis populations averaged 0.791, indicating relatively high genetic diversity. The results showed that the genetic differentiation pattern of A. nephrolepis was divided into two groups: Mt. Jungwangsan and other populations. Therefore, genetically diverse restoration materials can be secured from the Mt. Gariwangsan population to restore damaged areas. We found that individuals distributed within approximately 10 m were autocorrelated due to their high genetic similarity, whereas they were randomly distributed in the range of 15 m or more. The optimal sampling strategy involved collecting seeds from a minimum of 25 individuals within 10 m. This study suggests that genetic diversity in restoration material selection is necessary to enhance adaptability to environmental changes.

A typical molecular cloning procedure requires Sanger sequencing for sequence validation, which is cost-prohibitive and labor-intensive for large-scale clone analysis in genotype–phenotype studies. Here we present the cost-effective clone analysis platform TnClone, which uses next-generation sequencing based on Tn5 tagmentation to rapidly analyze a large number of clones from cell lysates. This method bypasses the extensive plasmid purification step. We also developed a user-friendly graphical user interface and provided general guidelines for conducting validation experiments. We tested our program with 1023 plasmids (222 from cell lysates and 801 from purified clones) and achieved 92% and 99.3% sensitivity with cell lysates and purified DNA, respectively. Our platform provides rapid turnaround with minimal hands-on time for secondary evaluation, as next-generation sequencing technology continues to evolve.

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