Genomic in situ hybridization on mitotic metaphase chromosomes of the HTL trispecies hybrid [(G. hirsutum × G. thurberi)2 × G. longicalyx)] and a BC2 progeny. A Mitotic metaphase showing 52 chromosomes of the HTL hybrid revealed by counterstaining with DAPI B mitotic metaphase showing in the HTL hybrid 13 green chromosomes from G. longicalyx, 13 yellow-orange chromosomes from the A-subgenome of G. hirsutum and 26 red chromosomes from G. thurberi and the D-subgenome of G. hirsutum revealed after the superimposition of FITC detection and Texas Red detection C mitotic metaphase showing 52 chromosomes in a HTL BC2 revealed by counterstaining with DAPI D mitotic metaphase in a HTL BC2 showing an entire chromosome of G. longicalyx (white arrow) and an intergenomic recombination (red arrow) showing movement of G. longicalyx chromatin into a chromosome of the A-subgenome of G. hirsutum. Scale bars: 5 μm.