Sugarcane is of important economic value for producing sugar and bioethanol. Tripidium arundinaceum (old name: Erianthus arundinaceum) is an intergeneric wild species of sugarcane that has desirable resistance traits for improving sugarcane varieties. However, the scarcity of chromosome markers has hindered the cytogenetic study of T. arundinaceum. Here we applied maize chromosome painting probes (MCPs) to identify chromosomes in sorghum and T. arundinaceum using a repeated fluorescence in situ hybridization (FISH) system. Sequential FISH revealed that these MCPs can be used as reliable chromosome markers for T. arundinaceum, even though T. arundinaceum has diverged from maize over 18 MYs (million years). Using these MCPs, we identified T. arundinaceum chromosomes based on their sequence similarity compared to sorghum and labeled them 1 through 10. Then, the karyotype of T. arundinaceum was established by multiple oligo-FISH. Furthermore, FISH results revealed that 5S rDNA and 35S rDNA are localized on chromosomes 5 and 6, respectively, in T. arundinaceum. Altogether, these results represent an essential step for further cytogenetic research of T. arundinaceum in sugarcane breeding.

The barcode probe is a convenient and efficient tool for molecular cytogenetics. Tripidium arundinaceum, as a polyploid wild allied genus of Saccharum, is a useful genetic resource that confers biotic and abiotic stress resistance for sugarcane breeding. Unfortunately, the basic cytogenetic information is still unclear due to the complex genome. We constructed the Cot-20 library for screening moderately and highly repetitive sequences from T. arundinaceum, and the chromosomal distribution of these repetitive sequences was explored. We used the barcode of repetitive sequence probes to distinguish the ten chromosome types of T. arundinaceum by fluorescence in situ hybridization (FISH) with Ea-0907, Ea-0098, and 45S rDNA. Furthermore, the distinction among homology chromosomes based on repetitive sequences was constructed in T. arundinaceum by the repeated FISH using the barcode probes including Ea-0663, Ea-0267, EaCent, 5S rDNA, Ea-0265, Ea-0070, and 45S rDNA. We combined these probes to distinguish 37 different chromosome types, suggesting that the repetitive sequences may have different distributions on homologous chromosomes of T. arundinaceum. In summary, this method provide a basis for the development of similar applications for cytogenetic analysis in other species.

Pennisetum purpureum is a potential species for biofuel production. Characterization and chromosomal distribution of retrotransposons could enhance the comprehension of the role and dynamics of the repetitive elements in plants. In this study, a phylogenetic tree was constructed according to the conserved reverse transcriptase sequences and revealed that these Ty1-copia retrotransposons had a typical structure. Analysis showed that the total Ty1-copia retrotransposons had a significant component, as high as 5.12 × 10³ copy numbers in P. purpureum. Then, the chromosomal pattern of four known lineages were also analyzed with the Pennisetum glaucum genome, which suggested that the Sire/Maximus lineage had the highest copy number and followed by Tork/Angela, Tork/TAR, Retrofit/Ale. Additionally, the chromosomal distribution of total Ty1-copia retrotransposons was detected by fluorescence in situ hybridization (FISH) to be a dispersed pattern with weak clustering, mostly near the centromeric regions of P. purpureum chromosomes; interestingly, there were four obvious signals in the subterminal chromosomes. These results suggested that there occurred differential dynamic evolution directions of Ty1-copia retrotransposons within P. purpureum. Furthermore, co-localization of Ty1-copia, 5S rDNA, and 35S rDNA indicated that two chromosome 2 and four chromosome 4 were identified. Concurrently, subterminal signals of Ty1-copia-type retrotransposons were located on four other homologous chromosomes. Altogether, these results shed light on the diversification of Ty1-copia retrotransposons and have the significance for generation of valid chromosomal markers in retrotransposon families.

Centromeres are essential for chromosome segregation, yet their repetitive nature makes them among the most difficult genomic regions to assemble and interpret. In polyploid plants, centromeres provide a unique record of chromosome evolution because centromeric repeats can be reshaped by polyploidization, subgenome differentiation and chromosomal rearrangement. Here, we combined low-coverage repeatome profiling, genome-wide mapping, fluorescence in situ hybridization (FISH), fiber-FISH and higher-order repeats (HORs) analysis to investigate centromere-associated repeats in Miscanthus sinensis (2n = 2x = 38). We identified MsSat1, a 137 bp satellite repeat representing approximately 2.1% of the genome, as a major centromere-associated satellite repeat. MsSat1 signals overlapped with the Saccharum officinarum centromeric retrotransposon probe and formed continuous tandem arrays on DNA fibers, supporting its association with centromeric domains. MsSat1-based clustering further recovered 13 related satellite variants, many of which showed strongly biased distribution between the A and B subgenomes. Among them, MsSat1-G displayed the most striking pattern, with strong B-subgenome enrichment and chromosome 7-specific localization. Detailed analysis revealed that MsSat1-G was embedded within an expanded MsSat1-rich domain on the fusion-derived chromosome 7, where it showed a more heterogeneous HORs architecture than the major MsSat1 array. These findings suggest that chromosome fusion was accompanied by local centromeric satellite amplification and repeat-architecture remodeling. Together, our study reveals that fusion-derived centromeres can preserve distinct satellite signatures and provides cytogenetic and genomic evidence that centromeric repeats record both subgenome differentiation and chromosome restructuring in M. sinensis.