Genomic in situ hybridization (GISH) was used to identify Festuca chromatin in mitotic chromosomes of Lolium multiflorum (Lm) × Festuca pratensis (Fp) hybrids and hybrid derivatives. In two inverse autoallotriploids LmLmFp and LmFpFp, in situ hybridization was able to discriminate between the Lolium and Festuca chromosomes. In a third triploid hybrid produced by crossing an amphiploid of L. multiflorum × F. pratensis (2n=4x=28) with L. multiflorum (2n=2x=14), the technique identified chromosomes with interspecific recombination. Also, in an introgressed line of L. multiflorum which was homozygous for the recessive sid (senescence induced degradation) allele from F. pratensis, a pair of chromosome segments carrying the sid gene could be identified, indicating the suitability of GISH in showing the presence and location of introgressed genes. By screening backcross progeny for the presence of critical alien segments and the absence of other segments the reconstitution of the genome of the recipient species can be accelerated.

The phylogeny of Festuca arundinacea Schreb. (2n = 6x = 42) was determined using GISH. Total genomic DNA of putative ancestral species was labelled with rhodamine and hybridized to chromosome preparations of hybrids involving these species and F. arundinacea. The degree of hybridization to chromosomes known to be homologous to the probe DNA was compared with that found simultaneously on chromosomes of the genome of F. arundinacea. It was concluded that the tetraploid species Festuca arundinacea var. glaucescens contributed two genomes and the diploid species Festuca pratensis one, to create the allohexaploid species F. arundinacea.Peer reviewe

Festuca, in situ hybridization, phylogeny, physical mapping, rDNA.

The position of the 18S-5.8S-26S and 5S rRNA genes have been physically mapped on the chromosomes of seven Lolium taxa. 18S-5.8S-26S sites were seen on two pairs of chromosomes in the inbreeding taxa. In the outbreeding taxa six sites were found in the L. multiflorum, seven in L. perenne and nine in L. rigidum var. rigidum. Two 5S sites were found in each of the taxa. In the inbreeders, the 5S sites were found adjacent to the 18S-5.8S-26S sites on chromosome 2. In L. multifiorum and L.perenne the 5S sites were on the short arm of chromosome 3. However, in L. rigidum var. rigidum the 5S rDNA site was found in either of the two positions.

Hybrids between T ambiguum and T repens were successfully established after ovule culture . The hybrids were male sterile but one of the hybrids produced a single seed when backcrossed to T repens, while the other produced a significantly greater number of seeds . The majority of the backcross hybrids from the latter had 48 chromosomes, being produced from the fertilisation of unreduced gametes of the F1 hybrid by T repens pollen . Regular meiosis in the BC1 hybrid resulted in the production of balanced gametes containing the full complement of T repens chromosomes and the polyhaploid set of T ambiguum chromosomes . There was no difficulty in establishing a substantial population of BC2 hybrids . The meiotic behaviour of the BC 1 hybrid indicated that it should be possible to transfer genes from T ambiguum into white clover T repens . The significance of the results in terms of the use of alien variation in the improvement of white clover is discussed .