The centromere has a conserved function across eukaryotes; however, the associated DNA sequences exhibit remarkable diversity in both size and structure. In plants, some species possess well-defined centromeres dominated by tandem satellite repeats and centromeric retrotransposons, while others have centromeric regions composed almost entirely of retrotransposons. Using a combination of bioinformatic, molecular, and cytogenetic approaches, we analyzed the centromeric landscape of Humulus lupulus. We identified novel centromeric repeats and characterized two types of centromeric organization. Cytogenetic localization on metaphase chromosomes confirmed the genomic distribution of the major repeats and revealed unique centromeric organization specifically on chromosomes 2, 8, and Y. Two centromeric types are composed of the major repeats SaazCEN and SaazCRM1 (Ty3/Gypsy) which are further accompanied by chromosome-specific centromeric satellites, Saaz40, Saaz293, Saaz85, and HuluTR120. Chromosome 2 displays unbalanced segregation during mitosis and meiosis, implicating an important role for its centromere structure in segregation patterns. Moreover, chromosome 2-specific centromeric repeat Saaz293 is a new marker for studying aneuploidy in hops. Our findings provide new insights into chromosome segregation in hops and highlight the diversity and complexity of the centromere organization in H. lupulus.

• Keywords
• Cannabaceae, asymmetric cell division, centromere, retrotransposons, sex chromosomes,
• MeSH
• Centromere * genetics   MeSH
• Chromosomes, Plant genetics   MeSH
• Humulus * genetics   MeSH
• Meiosis genetics   MeSH
• Repetitive Sequences, Nucleic Acid * genetics   MeSH
• Retroelements * genetics   MeSH
• Chromosome Segregation genetics   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Retroelements * MeSH

INTRODUCTION: Satellite DNA (satDNA) is a rapidly evolving component of plant genomes, typically found in (peri)centromeric, (sub)telomeric, and other heterochromatic regions. Due to their variability and species- or population-specific distribution, satDNA serves as valuable cytogenetic markers for studying chromosomal rearrangements and karyotype evolution among closely related species. Previous studies have identified species-specific subtelomeric repeats CS-1 in Cannabis sativa, HSR1 in Humulus lupulus, and HJSR in Humulus japonicus. These satellites have been used to differentiate sex chromosomes from autosomes, however, their evolutionary origins, sequence variation and conservation pattern across related species remain largely unexplored. METHODS: In this study, we analyze sequence similarity among these satellites and assess their interspecific chromosomal localization using fluorescence in situ hybridization (FISH). RESULTS: Our results reveal that the HSR1 and HJSR satellites are shared across all studied species, suggesting their common origin from a shared pool of satDNA in their common ancestor. In contrast, the CS-1 satellite exhibits higher sequence divergence. DISCUSSION: Although all three satellites are predominantly localized in subtelomeric regions, we identified species-specific exceptions. These findings provide new insight into the evolutionary dynamics of satDNA within the Cannabaceae family and offer further support for the divergence of Humulus species.

• Keywords
• Humulus, metaphase chromosomes, phylogenetics, satellite divergence, subtelomeric repeats,
• Publication type
• Journal Article MeSH