Nuclear expression of a cytoplasmic male sterility gene modifies mitochondrial morphology in yeast and plant cells

Duroc, Yann; Gaillard, Catherine; Hiard, Sophie; Tinchant, C.; Berthomé, Richard; Pelletier, Georges; Budar, Françoise

doi:10.1016/j.plantsci.2005.11.008

Cited by 33 publications

(24 citation statements)

References 58 publications

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“…Also, it is interesting to note that ORFH522 is reported to be lethal to bacterial cell (Nakai et al 1995). In the constructs, initially developed in pRT100 vector, CaMV 35S promoter was replaced with TA29 promoter for two reasons; one, 35S promoter is shown to effect a lower level expression in the male gametophyte in general and specifically in tapetal tissues of anther (Medberry et al 1992;Mc Cormick 2004;Duroc et al 2006) and two, to ensure formation of silencing trigger in the target tissue (tapetal cell layer) it was better to use the same promoter (TA29) employed in the male sterility inducing vector (Nizampatnam et al 2009) as has been the case in many of the other systems of transgenic male sterility and fertility restoration reported so far (Mariani et al 1992;Schmülling et al 1993;Zabaleta et al 1996;Busi et al 2006;Engelke et al 2010).…”

Section: Discussionmentioning

confidence: 99%

Intron hairpin and transitive RNAi mediated silencing of orfH522 transcripts restores male fertility in transgenic male sterile tobacco plants expressing orfH522

Nizampatnam

Kumar

2011

Plant Mol Biol

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The present work was aimed at developing vector construct(s) suitable for restoring fertility in transgenic male sterile tobacco plants expressing male-sterility-inducing ORFH522 in tapetal cell layer (Nizampatnam et al. Planta 229:987-1001, 2009). PTGS vectors that could produce either intron spliced hairpin RNA against the orfH522 or induce silencing of orfH522 by heterologous 3'UTR region were developed using the selected 316 bp (orf316) fragment of orfH522. The constructs were independently mobilized into Agrobacterium and used for transforming tobacco. The T(1) generation plants carrying the restorer gene cassettes in homozygous condition were identified and crossed with the male sterile transgenic tobacco plants to obtain the hybrid seeds. PCR analysis of hybrid plants indicated segregation for the sterility inducing cassette while all the plants carried the restorer cassette. Hybrid plants produced fertile pollen grains and formed normal capsules upon selfing. Further molecular analyses of these hybrid plants with RT-PCR, Northern blotting and siRNA detection, revealed that intron interrupted hairpin RNA (ihp-RNA) mediated gene silencing was more effective compared to silencing by heterologous 3'UTR (SHUTR) as indicated by the complete degradation of orfH522 transcripts and formation of higher levels of orf316 specific siRNA molecules in plants carrying ihp-RNA restorer construct. Segregation analyses of F(2) (selfed hybrid) plants confirmed the co-segregation of gene cassettes and the traits in Mendelian di-hybrid ratio (9:3:3:1). Taken together, the results established that intron hairpin and transitive RNAi mediated silencing of orfH522 transcripts restored fertility in transgenic male sterile tobacco plants expressing orfH522 and ihp-RNA was more efficient in silencing orfH522 transcripts.

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Section: Discussionmentioning

confidence: 99%

Intron hairpin and transitive RNAi mediated silencing of orfH522 transcripts restores male fertility in transgenic male sterile tobacco plants expressing orfH522

Nizampatnam

Kumar

2011

Plant Mol Biol

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“…However, genetic transformation of plant mitochondria is not yet technically feasible. Furthermore, we previously reported that in yeast, and most probably in plant cells, ORF138 does not correctly insert into the inner mitochondrial membrane when translated in the cytosol from a nuclear gene fusion with a mitochondrial targeting peptide sequence (Duroc et al 2006). Therefore, such approaches could not be undertaken.…”

Section: Discussionmentioning

confidence: 99%

The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria

et al. 2009

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The Ogura cytoplasmic male sterility causing protein, ORF138, was found to be part of a complex with an apparent size of over 750 kDa in the inner membrane of mitochondria of sterile plants. ORF138 did not colocalize with any of the oxidative phosphorylation complexes, nor did its presence modify their apparent size or amount, compared to samples from fertile isogenic plants. We attempted to detect potential proteins or nucleic acids that could be involved in the large ORF138 complex by 2D PAGE, immunoprecipitation and nuclease treatments of native extracts. All our results suggest that the ORF138 protein is the main, if not only, component of this large complex. The capacities of complexes I, II, IV, and ATP synthase were identical in samples from sterile and fertile plants. Isolated mitochondria from sterile plants showed a higher oxygen consumption than those from fertile plants. In vivo respiration measurements suggest that the difference in O(2) consumption measured at the organelle level is compensated at the cell/tissue level, completely in leaves, but only partially in male reproductive organs.

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“…Male sterile plants were obtained upon nuclear expression of the CMS-associated bean orf239 (He et al 1996), beet orf129 , chili pepper orf456 (Kim et al 2007), mustard orf220 , rice orf79 (Wang et al 2006) or wall-rocket orf108 (Kumar et al 2012). Conversely, similar attempts with the proteins associated with maize CMS-T, petunia CMS or radish and Brassica Ogura CMS failed to yield sterile plants (von Allmen et al 1991;Chaumont et al 1995;Wintz et al 1995;Duroc et al 2006). Also puzzling, in some cases, expression of the CMS orf resulted in aberrant pollen function and a semi-sterile or male-sterile phenotype regardless of the presence or not of an MTS (He et al 1996;Kumar et al 2012).…”

Section: Import Of Cms-associated Proteinsmentioning

confidence: 99%

“…When this gene, fused to an MTS-encoding sequence, was expressed in tobacco from a nuclear transgene, the URF13 protein actually accumulated in mitochondria, but there was no correlation with a male sterile phenotype (Chaumont et al 1995). The lack of male sterility following nuclear expression of several CMS genes (von Allmen et al 1991;Chaumont et al 1995;Wintz et al 1995;Duroc et al 2006;) is not clearly understood. Inappropriate timing and/or spatial control of transgene expression is often put forward as an explanation.…”

Section: Import Of Cms-associated Proteinsmentioning

confidence: 99%

Mitochondrial Genetic Manipulation

Mileshina

Niazi

Weber-Lotfi

et al. 2015

Somatic Genome Manipulation

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Nuclear expression of a cytoplasmic male sterility gene modifies mitochondrial morphology in yeast and plant cells

Cited by 33 publications

References 58 publications

Intron hairpin and transitive RNAi mediated silencing of orfH522 transcripts restores male fertility in transgenic male sterile tobacco plants expressing orfH522

Intron hairpin and transitive RNAi mediated silencing of orfH522 transcripts restores male fertility in transgenic male sterile tobacco plants expressing orfH522

The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria

Mitochondrial Genetic Manipulation

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