Karyotype analysis and classification of buckwheat chromosomes was performed with out chemical banding or staining using atomic force microscopy. F. esculentum and F. tartaricum chromosomes were isolated from the root tissues using enzymatic maceration technique and spread over a glass substrate. Air dried chromosomes had
a surface with ridges and the height of common and tartary buckwheat were approximately 350 and 150 nm.
Volumes of metaphase sets of buckwheat chromosomes were calculated using 3D atomic force microscopy
measurements. Chromosomes were morphologically characterized by the size, volume, arm lengths and ratios.
The calculated volumes of the F. esculentum and F. tartaricum chromosomes were in the ranges of 1.08 to 2.09 µm³ and 0.49 to 0.78 µm³ respectively. The parameters such as the relative arm length, centromere position and the chromosome volumes measured using AFM provides accurate karyomorphological classification by avoiding the subjective inconsistencies in banding patterns of conventional methods. The karyotype evolutionary trend indicates that F. esculentum is an ancient species compared to F. tartaricum. This is the first report of karyotyping of buckwheat using AFM.