Ogihara Y, Kurihara Y, Futami K, Tsuji K, Murai K
Kihara Institute for Biological Research, Yokohama City University, Japan.
Curr Genet. 1999 Dec;36(6):354-62. doi: 10.1007/s002940050510.
An alloplasmic wheat line with the cytoplasm of Aegilops crassa expresses photoperiod-sensitive cytoplasmic male sterility (PCMS). Southern- and Northern-hybridization analyses showed that this line contains alterations in both the gene structure and transcription patterns of the mitochondrial gene orf25. In this study, the nucleotide sequence around the orf25 gene of Ae. crassa (CR-orf25) and common wheat (AE-orf25) was determined, and we found that the upstream region of CR-orf25 had been replaced by that of rps7 of common wheat (AE-rps7) through recombination. A novel open reading frame (orf48) is present upstream of CR-orf25. In these three genes, transcription was initiated from the consensus promoter motif of plant mitochondrial genes located in the upstream regions. Processing enzymes in Ae. crassa and common wheat cleave the respective precursor mRNAs, namely CR-orf25 and AE-rps7, at sites similar to that of the premature mitochondrial 26S rRNA. In contrast, the precursor mRNA is not effectively processed at the target sequence of CR-orf25 in the alloplasmic wheat line. Because major transcripts of the euplasmic CR-orf25 and AE-rps7 genes would result in a truncated orf48 product, one possibility is that the orf48 protein might disturb mitochondrial function at a specific stage and hence affect the expression of the PCMS trait.
具有粗山羊草细胞质的异质小麦品系表现出光周期敏感细胞质雄性不育(PCMS)。Southern杂交和Northern杂交分析表明,该品系的线粒体基因orf25在基因结构和转录模式上均发生了改变。在本研究中,测定了粗山羊草(CR-orf25)和普通小麦(AE-orf25)中orf25基因周围的核苷酸序列,发现CR-orf25的上游区域通过重组被普通小麦的rps7(AE-rps7)的上游区域所取代。在CR-orf25的上游存在一个新的开放阅读框(orf48)。在这三个基因中,转录均起始于位于上游区域的植物线粒体基因的共有启动子基序。粗山羊草和普通小麦中的加工酶在类似于线粒体前体26S rRNA的位点切割各自的前体mRNA,即CR-orf25和AE-rps7。相反,在异质小麦品系中,前体mRNA在CR-orf25的靶序列处未得到有效加工。由于同质的CR-orf25和AE-rps7基因的主要转录本会产生截短的orf48产物,一种可能性是orf48蛋白可能在特定阶段干扰线粒体功能,从而影响PCMS性状的表达。