The fertility restorer genes X and T alter the transcripts of a novel mitochondrial gene implicated in CMS1 in chives (Allium schoenoprasum L.).
A chimeric mitochondrial gene configuration, mainly derived from sequences associated with the essential genes atp9 and atp6, was isolated from the sterility-inducing cytoplasm of the CMS1 system in chives (Allium schoenoprasum L.). This sequence is not found in four other cytoplasm types from chives; however, two copies are present in the mitochondrial DNA of CMS1-inducing cytoplasm, whose 5'-sequences are homologous to those of the atp9 gene. We provide evidence to show that one of the two CMS1-specific copies is actively transcribed, and two transcripts which terminate at the same position but differ in their 5'initiation sites were localized using the RACE technique. These transcripts of 942 and 961 nt, respectively, were confirmed to be the major products of this gene in CMS1 plants by Northern hybridization. However, smaller transcripts were found to accumulate in plants in which fertility had been restored. Restoration of fertility was induced either by the gene X, or the gene T at high temperatures. In (S1) X. genotypes a transcript with an estimated size of 440 nt was detected in all tissues examined. An additional hybridization signal with an estimated size of approximately 850 nt is expressed in temperature-sensitive plants [(S1) xxT.], and the intensity of a minor 350-nt transcript is enhanced. These latter alterations, conditioned by the gene T, occur independently of the growth temperature, but are limited to the flowers; they were not observed in leaves. The CMS1 transcripts are edited at seven positions and contain an ORF with a maximum coding capacity of 780 nt (containing the start codon derived from the atp9 gene in-frame). Use of the third in-frame start codon would result in the synthesis of a protein of a size very close to that of a previously described CMS1-specific protein, which has an apparent molecular weight of 18 kDa. The coding sequence that begins at this third in-frame start codon is also present in the sterility-inducing cytoplasms (S) and (T) in the onion, and absent in (N) cytoplasm.
Engelke
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Abteilung Angewandte Genetik, Universität Hannover, Herrenhäuser Str 2, 30419 Hannover, Germany. thomas.engelke@genetik.uni-hannover.de
Tatlioglu
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D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D027782:Chive / Q000235:genetics*; D003593:Cytoplasm / Q000235:genetics*; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism; D005298:Fertility / Q000235:genetics*; D017343:Genes, Plant / Q000235:genetics* / Q000502:physiology; D025261:Mitochondrial Proton-Translocating ATPases / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D011510:Proteolipids / Q000235:genetics; D006180:Proton-Translocating ATPases; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment
C063668:ATP9 protein, Arabidopsis; D029681:Arabidopsis Proteins; D017931:DNA Primers; D004272:DNA, Mitochondrial; D010940:Plant Proteins; D011510:Proteolipids; D012333:RNA, Messenger; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases