Complex organization of the mitochondrial genome of petaloid CMS carrot.

The petaloid trait in carrot is a mitochondrially associated homeotic-like conversion of stamens into petals which results in cytoplasmic male sterility (cms), but little is understood about the phenomenon at the molecular level. Identification of region(s) of the mitochndrial (mt) genome that are causally implicated in cms may be aided by a physical map of cms-associated mtDNA. The mt genome of petaloid cms carrot is 255 kb in length and contains three pairs of repeated sequences, two in direct orientation and one in inverted orientation. All regions of the genome are present in equal stoichiometries, but the arrangement of the repeated sequences prevented the representation of the entire genome as a single master circle or multiple subgenomes. An alternative model of mtDNA replication that accounts for our data is discussed.

Reduced symptoms of Verticillium wilt in transgenic tomato expressing a bacterial ACC deaminase.

Summary Ethylene evolved during compatible or susceptible disease interactions may hasten and/or worsen disease symptom development; if so, the prevention of disease-response ethylene should reduce disease symptoms. We have examined the effects of reduced ethylene synthesis on Verticillium wilt (causal organism, Verticillium dahliae) of tomato by transforming tomato with ACC deaminase, which cleaves ACC, the immediate biosynthetic precursor of ethylene in plants. Three promoters were used to express ACC deaminase in the plant: (i) CaMV 35S (constitutive expression); (ii) rolD (limits expression specifically to the site of Verticillium infection, i.e. the roots); and (iii) prb-1b (limits expression to certain environmental cues, e.g. disease infection). Significant reductions in the symptoms of Verticillium wilt were obtained for rolD- and prb-1b-, but not for 35S-transformants. The pathogen was detected in stem sections of plants with reduced symptoms, suggesting that reduced ethylene synthesis results in increased disease tolerance. The effective control of formerly recalcitrant diseases such as Verticillium wilt may thus be obtained by preventing disease-related ethylene production via the tissue-specific expression of ACC deaminase.

Widespread distribution of low-copy-number variants of mitochondrial plasmid pEM in the genus Agaricus.

The incidence of the linear mitochondrial plasmid pEM in Agaricus spp. was believed to be rare, based on visualization by gel electrophoresis and Southern hybridization. However, we report in this study PCR amplification of pEM-like sequences from all but one species of Agaricus examined. Regions amplified included (1) the pEM RNA polymerase gene and (2) adjoining carboxy-termini of the DNA and RNA polymerase genes. Sequence data from the RNA polymerase-like products support a plasmid, rather than mitochondrial, origin for these sequences. Sequence variation was low, and most differences were silent or conservative at the amino acid level. Stop codons were found in two of seven sequence types suggesting that functional constraints are low. A parsimony-derived phylogeny for these sequences did not match expected phylogenies for the host species. Recent acquisition of the plasmid is presented as the most likely hypothesis explaining these observations.

Plasmid RNA polymerase-like mitochondrial sequences in Agaricus bitorquis.

A linear mitochondrial plasmid, pEM, found in certain isolates of the basidiomycete Agaricus bitorquis, potentially encodes virus-like DNA and RNA polymerases. Mitochondrial DNA from Agaricus bisporus that hybridizes to an internal region of pEM contains a fragmented and potentially non-functional version of the carboxy terminal end of the plasmid RNA polymerase. In this study, we present the sequence of the corresponding region of mitochondrial DNA from A. bitorquis. This sequence contained the same region of the plasmid RNA polymerase gene as was reported for the mitochondrial DNA of A. bisporus, and the level of similarity between the A. bisporus and A. bitorquis mitochondrial sequences was much higher than the level of similarity between either mitochondrial sequence and the plasmid. We propose that this plasmid RNA polymerase-like sequence was present in the Agaricus mitochondrial genome before the divergence of A. bisporus and A. bitorquis, and thus is unlikely to be a recent derivative of the plasmid pEM.

Homology between mitochondrial DNA of Agaricus bisporus and an internal portion of a linear mitochondrial plasmid of Agaricus bitorquis.

Agaricus bisporus, the cultivated mushroom, contains a mitochondrial fragment (50H) which was previously demonstrated by Southern hybridization to have sequence similarity to an internal region of pEM, a linear mitochondrial plasmid of Agaricus bitorquis. The nucleotide sequence of 50H was determined and compared to the sequence of the corresponding pEM fragment. The region of sequence homology on pEM is contained within an open reading frame (ORF) that may encode an RNA polymerase, but 50H is neither an intact nor a complete copy of the ORF. pEM also contains an ORF with characteristics of genes for virus-encoded DNA polymerases. pEM appears to be very similar to other linear mitochondrial plasmids (in fungi and higher plants) reported to contain ORFs that may encode the same types of polymerases. The potential functionality of the pEM sequence suggests that it has diverged less than the mitochondrial fragment from a common ancestor.