Parental and novel copies of the mitochondrial orf25 gene in the hybrid crop-plant triticale: predominant transcriptional expression of the maternal gene copy.

Triticale, an intergeneric hybrid crop-plant, is generated when female wheat lines are fertilised with pollen from rye. We have investigated the mitochondrial DNA organisation and the expression of a total of 11 different triticale genotypes, varying in their nuclear and cytoplasmic backgrounds. In Southern hybridisations using probes homologous to the upstream flanking sequences, mtDNA fragments characteristic of both wheat and rye mtDNA can be detected in all triticale lines analysed. In addition, clones isolated fom a triticale lambda library exhibit either a maternal-like or paternal-like organisation of the orf25 gene region. By PCR cloning, four different orf25 gene copies were identified in triticale, three of which correspond to maternal (85%) or paternal (12%) orf25 sequences. Three percent of all clones represent a novel type, that might have arisen by homologous recombination. Although these data suggest biparental inheritance of mtDNA in wheat/rye crosses, paternal-like gene copies can also be detected in maternal wheat mitochondria. Their stoichiometry as assayed by competitive PCR is about 0.1% of total orf25 gene copies. The high abundance of paternal-like sequences in the F1 hybrid might therefore be due to either the transmission of rye mtDNA in the intergeneric cross and/or the amplification of sequences in triticale that persist in sub-stoichiometric amounts in wheat. These data suggest that amplification and recombination of sub-genomic mitochondrial molecules are affected by different nuclear genotypes. Interestingly, sequence analysis of triticale RT-PCR clones indicates a selective transcription of maternal-like orf25 gene copies in triticale. Mitochondrial gene expression may therefore possess mechanisms to compensate for the variation of mtDNA organisation.

Analysis of self-incompatibility interactions in 30 resynthesized Brassica napus lines. I. Fluorescence microscopic studies.

Thirty Brassica napus lines have been developed through interspecific hybridization of B. oleracea and B. campestris lines with defined S-allele constitutions. These lines, which represent 29 different S-allele combinations, were tested in a diallel of test-pollinations to determine the activity of the introgressed S-alleles and intergenomic dominance relationships. Some consistent trends were observed: B. oleracea S-alleles high in the dominance series (e.g. S8, S14, S29) were always active in the resynthesized B. napus lines, whereas recessive S-alleles (S2, S15) lost their activity in some test combinations. The B. campestris S-alleles were active in most cases, although 2 alleles were partially inactivated by the recessive B. oleracea allele S15.

Mitochondrial DNA of cytoplasmic male-sterile Triticum timopheevi: rearrangement of upstream sequences of the atp6 and orf25 genes.

The organization of mitochondrial DNA (mtDNA) and transcript patterns of the atp6 and orf25 genes were examined in cytoplasmic male-sterile (CMS) and fertile Triticum lines. Major differences are observed between CMS T. timopheevi and fertile T. aestivum for both mitochondrial genes. The T. aestivum mt genome carries two atp6 gene copies, whereas only a single copy of the atp6 gene is present in T. timopheevi mtDNA. Sequence data suggest that identical sequences upstream of the atp6 gene and the orf25 gene are involved in homologous recombination in both cytoplasms. The differences in the upstream sequences of the atp6 or the orf25 genes affect transcript sizes in both cytoplasms. Transcription initiation may occur at conserved promoter elements located at variable distances upstream of the aminoacid coding sequences. The correlation between the gene rearrangements and the CMS phenomenon in T. timopheevi is discussed.