Micropropagation is widely used in horticulture to provide large numbers of genetically similar, disease-free plants. The intent is to produce high-quality genetically identical planting material to ensure uniformity. However, this method can result in somaclonal variation, which refers to the increased rate of unusual phenotypic developments that result from genetic mutations or epigenetic alterations. While some mutations can be visually identified due to morphological or physiological defects, many are difficult to assess based on observation alone. Various DNA markers have been employed to assess the genetic fidelity of plants. However, these techniques have limited genome coverage, which makes it challenging to determine the overall genetic fidelity of the clones. Despite this, markers have widely been employed to assess genetic fidelity and to make claims that the process produces true-to-type plants. Genotyping-by-sequencing (GBS) can sequence and genotype thousands of markers from multiple individuals simultaneously and is now a common and low-cost technique. In this study, the efficacy of GBS versus simple sequence repeats (SSRs) in evaluating genetic fidelity in a clonal population of micropropagated cannabis plants was compared. The SSR analysis resulted in eight readable markers indicating negligible genetic diversity. In contrast, the GBS analysis identified over 9000 variants in cannabis clones, revealing genetic diversity within clonal lines across multiple subcultures. This study demonstrated the increased precision of sequencing-based technologies for detecting somaclonal mutations and highlighted the limitations of using SSRs or similar markers to evaluate genetic fidelity in these systems.

微繁殖广泛应用于园艺，以提供大量遗传相似、无病害的植物。目的是生产高质量的基因相同的种植材料，以确保一致性。然而，这种方法可能会导致体细胞克隆变异，这是指由于基因突变或表观遗传改变而导致的异常表型发育率增加。虽然由于形态或生理缺陷，一些突变可以通过肉眼识别，但许多突变很难仅根据观察进行评估。各种 DNA 标记已被用来评估植物的遗传保真度。然而，这些技术的基因组覆盖范围有限，这使得确定克隆的整体遗传保真度具有挑战性。尽管如此，标记已广泛用于评估遗传保真度并声称该过程产生了真实的植物。测序基因分型 (GBS) 可以同时对多个个体的数千个标记进行测序和基因分型，现在是一种常见且低成本的技术。在本研究中，GBS 的功效与简单序列重复（SSR）在评估微繁殖大麻植物克隆群体的遗传保真度方面进行了比较。SSR 分析产生了八个可读标记，表明遗传多样性可以忽略不计。相比之下，GBS 分析在大麻克隆中发现了 9000 多个变异，揭示了多个亚文化的克隆系内的遗传多样性。这项研究证明了基于测序的技术用于检测体细胞克隆突变的精度有所提高，并强调了使用 SSR 或类似标记来评估这些系统中遗传保真度的局限性。