Laser B, Mohr S, Odenbach W, Oettler G, Kück U
Lehrstuhl für Allgemeine Botanik, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Curr Genet. 1997 Nov;32(5):337-47. doi: 10.1007/s002940050286.
Triticale, an intergeneric hybrid crop-plant, is generated when female wheat lines are fertilised with pollen from rye. We have investigated the mitochondrial DNA organisation and the expression of a total of 11 different triticale genotypes, varying in their nuclear and cytoplasmic backgrounds. In Southern hybridisations using probes homologous to the upstream flanking sequences, mtDNA fragments characteristic of both wheat and rye mtDNA can be detected in all triticale lines analysed. In addition, clones isolated fom a triticale lambda library exhibit either a maternal-like or paternal-like organisation of the orf25 gene region. By PCR cloning, four different orf25 gene copies were identified in triticale, three of which correspond to maternal (85%) or paternal (12%) orf25 sequences. Three percent of all clones represent a novel type, that might have arisen by homologous recombination. Although these data suggest biparental inheritance of mtDNA in wheat/rye crosses, paternal-like gene copies can also be detected in maternal wheat mitochondria. Their stoichiometry as assayed by competitive PCR is about 0.1% of total orf25 gene copies. The high abundance of paternal-like sequences in the F1 hybrid might therefore be due to either the transmission of rye mtDNA in the intergeneric cross and/or the amplification of sequences in triticale that persist in sub-stoichiometric amounts in wheat. These data suggest that amplification and recombination of sub-genomic mitochondrial molecules are affected by different nuclear genotypes. Interestingly, sequence analysis of triticale RT-PCR clones indicates a selective transcription of maternal-like orf25 gene copies in triticale. Mitochondrial gene expression may therefore possess mechanisms to compensate for the variation of mtDNA organisation.
小黑麦是一种属间杂交作物,由小麦雌性品系与黑麦花粉受精产生。我们研究了11种不同小黑麦基因型的线粒体DNA组织情况及其表达,这些基因型在核背景和细胞质背景方面存在差异。在使用与上游侧翼序列同源的探针进行的Southern杂交中,在所有分析的小黑麦品系中都能检测到小麦和黑麦线粒体DNA特有的线粒体DNA片段。此外,从小黑麦λ文库中分离出的克隆在orf25基因区域呈现出母本样或父本样的组织形式。通过PCR克隆,在小黑麦中鉴定出了四种不同的orf25基因拷贝,其中三种对应于母本(85%)或父本(12%)的orf25序列。所有克隆的3%代表一种新型,可能是通过同源重组产生的。尽管这些数据表明在小麦/黑麦杂交中mtDNA存在双亲遗传,但在母本小麦线粒体中也能检测到父本样基因拷贝。通过竞争性PCR测定,它们的化学计量约为总orf25基因拷贝的0.1%。因此,F1杂种中父本样序列的高丰度可能是由于属间杂交中黑麦线粒体DNA的传递和/或小黑麦中序列的扩增,这些序列在小麦中以亚化学计量的量持续存在。这些数据表明亚基因组线粒体分子的扩增和重组受不同核基因型的影响。有趣的是,小黑麦RT-PCR克隆的序列分析表明,小黑麦中母本样orf25基因拷贝存在选择性转录。因此,线粒体基因表达可能具有补偿mtDNA组织变异的机制。