One of the most popular, cheap and widely used approaches in comparative cytogenetics - especially by botanists - is that concerning intrachromosomal and interchromosomal karyotype asymmetry. Currently, there is no clear indication of which method, among the many different ones reported in literature, is the most adequate to infer karyotype asymmetry (especially intrachromosomal), above all in view of the criticisms recently moved to the most recent proposal published. This work addresses a critical review of the methods so far proposed for estimation of karyotype asymmetry, using both artificial and real chromosome datasets. It is shown once again how the concept karyotype of asymmetry is composed by two kinds of estimation: interchromosomal and intrachromosomal asymmetries. For the first one, the use of Coefficient of Variation of Chromosome Length, a powerful statistical parameter, is here confirmed. For the second one, the most appropriate parameter is the new Mean Centromeric Asymmetry, where Centromeric Asymmetry for each chromosome in a complement is easily obtained by calculating the difference of relative lengths of long arm and short arm. The Coefficient of Variation of Centromeric Index, strongly criticized in recent literature, is an additional karyological parameter, not properly connected with karyotype asymmetry. This shows definitively what and how to measure to correctly infer karyotype asymmetry, by proposing to couple two already known parameters in a new way. Hopefully, it will be the basic future reference for all those scientists dealing with cytotaxonomy.
Keywords: Artificial chromosome datasets; Stebbins classification; asymmetry indices; chromosomal heterogeneity; interchromosomal asymmetry; intrachromosomal asymmetry; karyological parameters; karyotype asymmetry.