Integrated Optimization of Seed Germination, Cytogenetic Techniques, and Digital Image Analysis for Karyotype construction in Selected Plant Species from North Cyrenaica, Libya
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Seed Germination, Cytogenetic, Karyotype Analysis, Chromosome Visualization, Digital Image Analysis, Libyan Flora
Abstract
Seed germination and chromosome preparation are fundamental steps in cytogenetic studies, particularly in plant species with seed dormancy and specific germination requirements. In North Cyrenaica, Libya, cytogenetic information remains limited for many native and endemic plant species. The aim of this study was to optimize seed germination, chromosome preparation, and digital image analysis for karyotype construction in three plant species from North Cyrenaica, Libya: Arum cyrenaicum, Arbutus pavarii, and Origanum cyrenaicum. Different germination treatments were evaluated to obtain suitable root tips for chromosome analysis. Species-specific procedures were developed through the optimization of pretreatment, fixation, hydrolysis, staining, and slide preparation steps. Several image processing software tools were also assessed to improve chromosome visualization and karyogram construction. The three species exhibited different germination responses, reflecting variation in dormancy mechanisms and environmental requirements. The optimized procedures produced clear metaphase chromosomes suitable for chromosome counting and karyological analysis. Digital image analysis enhanced chromosome visualization and facilitated individual chromosome identification, homologous chromosome pairing, and karyogram construction. The developed workflow offers a reliable approach for seed germination and cytogenetic studies of native and endemic plant species and provides valuable baseline data for future taxonomic and evolutionary research on the Libyan flora.
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