Evaluation of the efficiency of sexual reproduction in restoring Podospora anserina mitochondrial DNA to wild-type.

A Podospora anserina mitochondrial DNA (mtDNA) rearrangement mutant, Mn19, was crossed with a deletion mutant, alphaDelta5. Ascospores (212) from random asci were tested for viability, growth and life-span phenotypes, and mtDNA inheritance. Some spore inviability was detected along with early growth arrest (at the time of spore germination) from which some isolates recovered. However, the majority had wild-type growth and life-span phenotypes. All isolates tested at the DNA level (102) had wild-type mtDNA hybridization patterns with probes that detected defects in the parents. About 20% also inherited low levels of mtDNA molecules with the rearrangement characteristic of the Mn19 parent. These results demonstrate that P. anserina has a remarkable ability, through sexual reproduction, to restore its mtDNA to wild-type, even when the parents are predominately mutant.

Elimination of mitochondrial mutations by sexual reproduction: two Podospora anserina mitochondrial mutants yield only wild-type progeny when mated.

In order to understand the transmission of mitochondrial mutations in sexual crosses of Podospora, we attempted to create compatible strains with defined mitochondrial mutations. A previously characterized mutant, Mn19, with a bipartite mitochondrial genome, served as the fertilizing parent in a cross with a mitochondrial deletion mutant, alphadelta5. Characterization of the deletion mutant is reported here. All six of the monokaryotic progeny isolated had neither parental defect but instead appeared to have inherited wild-type mitochondrial DNA. One of the progeny had a mitochondrial plasmid derived from intramolecular recombination between an 11-bp repeated mitochondrial sequence. Subsequent analysis using the polymerase chain reaction (PCR) identified rare undeleted wild-type mtDNA sequences in the maternal parent. The uniform inheritance of wild-type mitochondrial DNA suggests either an aggressive repair mechanism or else selective amplification and transmission of rare wild-type mtDNA molecules.

Genetic and molecular analysis of a long-lived strain of Podospora anserina.

A genetic and molecular analysis of a long-lived strain of Podospora anserina, Mn19, was undertaken to detect mutations in genes responsible for senescence. In crosses between Mn19 and wild type about 15% of the progeny were long-lived, regardless of the female parent. Molecular analysis of the long-lived progeny showed that none of the strains inherited a mtDNA rearrangement characteristic of the Mn19 parent. Instead, all long-lived strains initially inherited wild-type mtDNA. Over time the mtDNA of most long-lived strains underwent rearrangements, deletions and amplifications. The change over time in the presence of two previously characterized plasmids associated with either senescence or longevity was monitored. Crosses between Mn19 and its long-lived progeny also yielded only a small percent of individuals recovering from senescence. Analysis of mtDNA from crosses suggests that wild-type mtDNA from the paternal parent can be selected over mtDNA from the maternal parent. The life span phenotypes of progeny were not consistent with the hypothesis that mutations in a few nuclear genes were responsible for longevity.

A mitochondrial DNA rearrangement and three new mitochondrial plasmids from long-lived strains of Podospora anserina.

The excision-junction sites of a mtDNA rearrangement of a long-lived strain of Podospora anserina, Mn19, were cloned and sequenced. Analysis of sequence and hybridization data lead to the conclusion that the Mn19 mtDNA consists of two nonoverlapping circular molecules. Three plasmids, LMt-2, LMt-3, and LMt-4, cloned from long-lived progeny of crosses between the Mn19 strain and wild type were cloned and sequenced. These plasmids share features and excision-junction sites with previously described longevity and senescence plasmids. The Mn19 mtDNA rearrangement and plasmids LMt-2, LMt-3, and LMt-4 are described. The possible significance of similarities to previously described plasmids is discussed.

Non-mendelian inheritance of mitochondrial DNA and ribosomal DNA in the myxomycete, Didymium iridis.

The inheritance of both the mitochondrial DNA (mtDNA) and the nuclear-encoded extrachromosomal ribosomal DNA (rDNA) has been studied in the myxomycete, Didymium iridis, by DNA-DNA hybridization of labeled probes to total DNA at various stages of the life cycle. Both the mtDNA and rDNA populations rapidly become homogeneous in individuals, but there is a qualitative difference in the patterns of inheritance of these two molecules. One parental rDNA type was preferentially inherited in all crosses; selective replication of this molecule is tentatively proposed as the mechanism of inheritance. In contrast, either parental mtDNA type could be inherited. Since the inherited population of parental mtDNA molecules are not partitioned into cells in this coenocytic organism, no known mechanism of inheritance can explain the rapid and apparently random loss of one parental mtDNA type in individuals.