C. S. Levings

Mitochondrial DNA in the T male-sterile cytoplasm (cms-T) of maize contains an open reading frame (ORF 13) associated with the T type of sterility. Antibodies raised to a chemically synthesized oligopeptide corresponding to ORF 13 were used to establish the expression of a 13-kDa protein from this reading frame. The 13-kDa polypeptide is synthesized uniquely in cms-T maize and purifies with the membrane fraction of T mitochondria. We assign the symbol urf13-T to designate this mitochondrial gene. Presence of the nuclear restorer gene Rf1 in cms-T plants results in a decrease in abundance of 13-kDa protein and alteration in the transcripts of urf13-T.

The Texas cytoplasm of maize carries two cytoplasmically inherited traits, male sterility and disease susceptibility, which have been of great interest both for basic research and plant breeding. The two traits are inseparable and are associated with an unusual mitochondrial gene, T-urf13, which encodes a 13-kilodalton polypeptide (URF13). An interaction between fungal toxins and URF13, which results in permeabilization of the inner mitochondrial membrane, accounts for the specific susceptibility to the fungal pathogens.

Mitochondrial DNAs were prepared from maize lines with normal cytoplasm and with the T, C, S, and EP sources of male-sterile cytoplasms. Agarose gel electrophoresis of these preparations revealed a main high-molecular-weight DNA band. In addition, the S cytoplasm was characterized by the presence of two faster migrating DNAs of molecular weight 3.42 to 3.48 x 10(6) and 4.01 to 4.10 x 10(6). Electron microscopy showed these unique DNAs to be of different length, but their molecular configuration was not clearly established. It is possible that these unique DNAs represent physical evidence of an episomal system previously postulated to function in the S male-sterile cytoplasm.

Maize mitochondrial and chloroplast DNA's were prepared from normal (fertile) lines or single crosses and from members of the T, C, and S groups of male-sterile cytoplasms. Restriction endonucleases HindIII, BamI, EcoRI, and SalI were used to restrict the DNA, and the resultant fragments were electrophoresed in agarose gels. The results show that the N (fertile), T, C, and S cytoplasms each contained distinct mitochondrial DNA (mtDNA). These distinctive patterns were unaffected by nuclear genotype. No evidence of paternal inheritance of mtDNA was observed. Chloroplast DNA (ctDNA) from the N, C, and T cytoplasms was indistinguishable by HindIII, SalI, or EcoRI endonuclease digestion. The S cytoplasm ctDNA, however, was slightly different from that of other cytoplasms, as indicated by a slight displacement of one band in HindIII digests. The molecular weight of maize ctDNA was estimated to be as high as 88 x 10(6). Estimates of the minimum molecular weight of maize mtDNA ranged from 116-131 x 10(6), but the patterns were to complex for an unambiguous determination. Based on HindIII data, a comparison of the molecular weight of mtDNA bands common to the N, T. C, and S cytoplasms suggests that C cytoplasm most closely resembles N cytoplasm. The T and S sources are more divergent from the C and N cytoplasms. These results indicate a possible gradation of relatedness among male-sterile cytoplasms. The marked variation in mtDNA, with apparently less variation in ctDNA, represents circumstantial, but compelling, evidence that mtDNA may be involved in the male sterility and disease susceptibility traits in maize.