Molecular cytogenetic (FISH) and genome analysis of diploid wheatgrasses and their phylogenetic relationship.

This paper reports detailed FISH-based karyotypes for three diploid wheatgrass species Agropyron cristatum (L.) Beauv., Thinopyrum bessarabicum (Savul.&Rayss) A. Löve, Pseudoroegneria spicata (Pursh) A. Löve, the supposed ancestors of hexaploid Thinopyrum intermedium (Host) Barkworth & D.R.Dewey, compiled using DNA repeats and comparative genome analysis based on COS markers. Fluorescence in situ hybridization (FISH) with repetitive DNA probes proved suitable for the identification of individual chromosomes in the diploid JJ, StSt and PP genomes. Of the seven microsatellite markers tested only the (GAA)n trinucleotide sequence was appropriate for use as a single chromosome marker for the P. spicata AS chromosome. Based on COS marker analysis, the phylogenetic relationship between diploid wheatgrasses and the hexaploid bread wheat genomes was established. These findings confirmed that the J and E genomes are in neighbouring clusters.

Constructing an alternative wheat karyotype using barley genomic DNA.

The established karyotype was generated by genomic in situ hybridization (GISH) using total barley genomic DNA as labelled probe on mitotic metaphase bread wheat chromosomes. GISH produced specific banding signals on 16 of the 21 chromosome pairs. The following chromosomes showed distinctive banding patterns: 2A, 3A, 4A, 5A, 6A, 7A, 1D, 2D, 7D and all of the B chromosomes. The remaining chromosomes showed either faint bands or no hybridization signals at all. The in situ hybridization patterns corresponded to the GAA-satellite sequence, which is similar to the N-banding pattern in wheat. In situ hybridization by labelling total barley genomic DNA made it possible to identify most of the bread wheat chromosomes. The present paper describes a GISH-banding method for hexaploid wheat chromosomes. It is a valuable alternative method for fast chromosome selection without using FISH repetitive DNA clones.