Homology-based cloning and expression analysis of Rf genes encoding PPR-containing proteins in tobacco
文献类型: 外文期刊
作者: Ding, A. M.;Li, F. X.;Chen, Y. Q.;Zong, P.;Qu, X.;Gong, D. P.;Liu, G. S.;Sun, Y. H.;Ding, A. M.;Chen, Y. Q.;Zong, P.;Qu, X.
作者机构:
关键词: Rf gene;Pentatricopeptide repeat protein family;Quantitative reverse transcription polymerase chain reaction;Sequence structure analysis;Phylogenetic tree
期刊名称: GENETICS AND MOLECULAR RESEARCH
ISSN: 1676-5680
年卷期: 2014 年 13 卷 1 期
页码:
收录情况: SCI
摘要: As a model plant, mechanisms of the cytoplasmic male sterility/restoration of fertility (CMS/Rf) system in tobacco are seldom studied. Using Rf gene sequences from other Solanaceae plants and the draft genome of Nicotiana benthamiana, degenerate primers were designed to amplify the cDNA pool of N. tomentosiformis. In total, six possible Rf sequences were identified, two of which contained base-deletion mutations. The other four were intact open reading frames, of which NtomPPR5 harbored a 3-pentatricopeptide repeat (PPR) motif deletion. Structure analysis revealed that they all encoded a PPR-containing protein with putative mitochondrial targeting signals at their N-terminus, and they all belong to the P subfamily. Phylogenetic analysis showed that all of the Rf-coding PPRs clustered together, and recent duplication events might have occurred in tobacco after the divergence of the species. Quantitative reverse transcription polymerase chain reaction analysis demonstrated that the NtomRfs were expressed in all tissues of N. tomentosiformis and (CMS) K326, although the expression levels varied with gene, organ, and developmental stage. Furthermore, the expression levels of Rf sequences in K326 were lower than those in CMS K326. The molecular basis of the CMS/Rf system in tobacco requires further investigation.
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