Physical and topographical mapping among Triticeae chromosomes.

Three principal approaches have been used in our laboratory to analyze Triticeae genomes. (i) Synteny analysis: synteny among different Gramineae genomes was studied employing the elegant system of the Agropyron chromosome-induced deletion lines of wheat. Deletion mapping, predominantly of the homoeologous group 7 chromosomes, has led to the construction of a high density physical consensus map of wheat. The integration of wheat, barley and oat RFLP markers proves the colinearity between the wheat A-, B- and D-genomes, the H-genome of barley, and the E-genome of Agropyron. (ii) Light microscopic in situ techniques: the recent improvement of a drop technique for plant protoplasts was crucial for the sensitivity enhancement of fluorescence in situ hybridization (FISH), the efficient preparation of plant chromosomes for high resolution scanning electron microscopy, mapping of low-copy sequences, and comparative in situ hybridization. A tandemly amplified repetitive sequence element from microdissected barley chromosomes has enabled the karyotyping of Gramineae genomes in a single step. We have isolated and characterized members of this element family from other Triticeae species using PCR. The significant interspecific sequence differences were useful to identify single plant genomes, chromosomes and chromosome segments via post-hybridization washes under different stringency conditions. These sequences are also useful for simultaneous double or triple hybridization experiments in an attempt to localize new sequences on specific chromosomes or chromosome segments. The physical mapping of the Sec-1 locus has been refined on the satellite of chromosome 1R of rye, and the syntenic locus on barley chromosome 1H was identified. (iii) Physical mapping of rDNA sequences by high resolution electron microscopy: a method was developed for in situ hybridization and signal detection using high resolution field emission scanning electron microscopy and a backscattered electron detector. Colloidal gold particles were localized on chromosome structures resembling the 30 nm fibre. An rDNA probe was located in the secondary constriction and the highly compact adjacent regions of barley chromosomes.