Structural and functional analysis of mitochondrial plasmids in Claviceps purpurea.

Several strains of Claviceps purpurea, a phytopathogenic Ascomycete, contain mitochondrial (mt) plasmids in high molar excess relative to mtDNA. Comparative analysis of plasmids of four strains of different geographic origin revealed that all the plasmids are structurally related (size; linearity; restriction map; probably 5'-linked terminal protein; terminal inverted repeats, TIRs); two of them are even identical, indicating a possible mobility of these genetic entities. In strain K it was shown that plasmid titres are comparably high in axenic cultures and in parasitic structures (sclerotia). Detailed analysis of plasmid pClK1 proved the existence of a perfect TIR of 327 bp; the plasmid's structure and details of its nucleotide sequence indicate a replication modus comparable to that of adenoviruses. pClK1 is almost completely transcribed resulting in two major transcripts of 3.5 and 3.15 kb, respectively. In plasmid-free strains (cured by ethidium bromide treatment) these mRNAs are not detectable; nevertheless they show no significant difference in phenotype. As judged from their structural properties they could be derived from viral ancestors. In this context the plasmids' close relationship to mt plasmids of higher plants may be of special interest.

Extrachromosomal genetics of Claviceps purpurea : I. Mitochondrial DNA and mitochondrial plasmids.

In order to establish a system for gene cloning in the ergot fungus Claviceps purpurea mitochondrial DNA was purified from two wild strains and four descendants of production strains. A characterization of the mt DNA with respect to buoyant density, molecular weight (45 kb corresponding to acontour lenght of 14 µm) showed no differences. However, considerable differences were detected in the restriction pattern, with the exception of two commercial strains of possibly common origin. From this it follows that restriction patterns of mt DNA may be used as finger prints to identify related strains.In one of the wild strains plasmids associated with the mitochondria were found. A characterization of two species p11 and p12 having molecular weights of 6.6 and 5.3 kb revealed that these genetic units are linear having respective contour lengths of 2.1 and 1.7 µm. According to Southern blot analyses both plasmids show homology to each other, but not to the mitochondrial "chromosome". Accordingly, p11 and p12, despite localization in the mitochondria, are evidently not an integral part of the mt "chromosome". The possibility to use the mt DNA and mitochondrial plasmids to establish a vector system for molecular cloning in imperfect strains of Claviceps purpurea has been pointed out.