Comparative karyotype analysis among cytotypes of Cynodon dactylon (L.) Pers. (Poaceae)

Cynodon dactylon is characterized by taxonomic and systematic complexity, and polyploidy is one of the factors responsible for its genetic and morphological diversity. The aim of the present study was to compare karyotypes of C. dactylon cytotypes based on fluorescent banding and nuclear DNA content. The nine C. dactylon accessions evaluated were obtained from the Active Germplasm Bank (BAG) of the United States Department of Agriculture (USDA). Roots were pretreated with cycloheximide, fixed in Carnoy's solution and subjected to enzymatic digestion. Slides were prepared by the dissociation and air-drying technique. The fluorescent banding pattern was obtained using chromomycin A3 (CMA)/4,6-dimidino-2-phenylindole (DAPI) staining and DNA content was estimated by flow cytometry. The chromosome number of the accessions ranged from 2n = 2x = 18 to 2n = 5x = 45. Chromosomal polymorphism was observed based on the distribution and number of heterochromatic bands, with CMA⁺ bands located in the pericentromeric position and DAPI⁺ bands mainly in the terminal position. PI477004-26 (2n = 3x = 27) and PI291966-27 (2n = 4x = 36) had the highest and lowest number of DAPI⁺ bands, respectively. The number of CMA⁺ bands was stable, as only PI477004-26, PI291966-27 and PI289750-10 (2n = 5x = 45) showed variation. There was no direct correlation between an increase in the ploidy level and an increase in the percentage of heterochromatic regions, mainly in relation to A-T-rich blocks. The chromosomal banding variation found reinforces the notion of allopolyploidy occurrence in C. dactylon and demonstrates the genomic complexity of this species regard to repetitive DNA content.