Use of an inducible reporter gene system for the analysis of auxin distribution in the moss Physcomitrella patens

Bierfreund, Nicole M.; Reski, Ralf; Decker, Eva L.

doi:10.1007/s00299-003-0646-1

Cited by 83 publications

(76 citation statements)

References 29 publications

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“…In situ immunolocalization experiments showed that actively proliferating apices of the filaments did have higher concentrations of IAA. Similar results have been obtained in filaments of the bryophyte Physcomitrella patens, where the expression of GH3::GUS and DR5::GUS transgenes indicates higher concentrations of auxin in the young, actively growing cells of the protonemal filaments (Bierfreund et al, 2003), similarly located at the apices (Cove et al, 2006). Therefore, a possible scenario in agreement with the data from the ablation experiment is that, in an intact filament, there is an IAA gradient that reaches its maximum at both extremities of the filament, where E cells are located.…”

Section: Role Of Auxin In E Siliculosus Developmentsupporting

confidence: 71%

Auxin Metabolism and Function in the Multicellular Brown AlgaEctocarpus siliculosus

et al. 2010

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Ectocarpus siliculosus is a small brown alga that has recently been developed as a genetic model. Its thallus is filamentous, initially organized as a main primary filament composed of elongated cells and round cells, from which branches differentiate. Modeling of its early development suggests the involvement of very local positional information mediated by cell-cell recognition. However, this model also indicates that an additional mechanism is required to ensure proper organization of the branching pattern. In this paper, we show that auxin indole-3-acetic acid (IAA) is detectable in mature E. siliculosus organisms and that it is present mainly at the apices of the filaments in the early stages of development. An in silico survey of auxin biosynthesis, conjugation, response, and transport genes showed that mainly IAA biosynthesis genes from land plants have homologs in the E. siliculosus genome. In addition, application of exogenous auxins and 2,3,5-triiodobenzoic acid had different effects depending on the developmental stage of the organism, and we propose a model in which auxin is involved in the negative control of progression in the developmental program. Furthermore, we identified an auxin-inducible gene called EsGRP1 from a small-scale microarray experiment and showed that its expression in a series of morphogenetic mutants was positively correlated with both their elongated-to-round cell ratio and their progression in the developmental program. Altogether, these data suggest that IAA is used by the brown alga Ectocarpus to relay cell-cell positional information and induces a signaling pathway different from that known in land plants.

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Section: Role Of Auxin In E Siliculosus Developmentsupporting

confidence: 71%

Auxin Metabolism and Function in the Multicellular Brown AlgaEctocarpus siliculosus

et al. 2010

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“…In order to discriminate between these alternatives, we compared the responses of GmGH3pro:GUS and PpSHI2pro:PpSHI2-GUS lines to exogenous auxin. As previously reported (Bierfreund et al, 2003;Fujita et al, 2008), the GmGH3 promoter was induced by auxin treatment in a concentrationdependent manner, which resulted in an almost uniform staining of the entire gametophore, suggesting that all cell types can respond to auxin (Decker et al, 2006) (Fig. 2).…”

Section: Ppshi1/2 Expression Coincides With Sites Of Auxin Responsementioning

confidence: 51%

“…The PpSHI1/2 expression pattern in the adult gametophore, with maxima in rhizoids and axillary hairs, is almost identical to that of a GmGH3pro:GUS reporter [Gm, Glycine max (soybean)] used to detect sites of auxin response (Bierfreund et al, 2003;Fujita et al, 2008). If not coincidental, this correlation could indicate that the PpSHI genes are induced by auxin, or that their expression is a prerequisite for the generation of auxin peaks or auxin responsiveness.…”

Section: Ppshi1/2 Expression Coincides With Sites Of Auxin Responsementioning

confidence: 85%

Homologues of the Arabidopsis thaliana SHI/STY/LRP1 genes control auxin biosynthesis and affect growth and development in the moss Physcomitrella patens

et al. 2010

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SUMMARYThe plant hormone auxin plays fundamental roles in vascular plants. Although exogenous auxin also stimulates developmental transitions and growth in non-vascular plants, the effects of manipulating endogenous auxin levels have thus far not been reported. Here, we have altered the levels and sites of auxin production and accumulation in the moss Physcomitrella patens by changing the expression level of homologues of the Arabidopsis SHI/STY family proteins, which are positive regulators of auxin biosynthesis genes. Constitutive expression of PpSHI1 resulted in elevated auxin levels, increased and ectopic expression of the auxin response reporter GmGH3pro:GUS, and in an increased caulonema/chloronema ratio, an effect also induced by exogenous auxin application. In addition, we observed premature ageing and necrosis in cells ectopically expressing PpSHI1. Knockout of either of the two PpSHI genes resulted in reduced auxin levels and auxin biosynthesis rates in leafy shoots, reduced internode elongation, delayed ageing, a decreased caulonema/chloronema ratio and an increased number of axillary hairs, which constitute potential auxin biosynthesis sites. Some of the identified auxin functions appear to be analogous in vascular and non-vascular plants. Furthermore, the spatiotemporal expression of the PpSHI genes and GmGH3pro:GUS strongly overlap, suggesting that local auxin biosynthesis is important for the regulation of auxin peak formation in non-vascular plants.

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“…A tetracyclinerepressible gene-expression system has been reported in P. patens (Zeidler et al, 1996), but it requires continuous growth on tetracycline, which is detrimental to the moss (Laloue M., personal communication). Abscissic acid or auxin-induced gene expression have also been shown using, respectively, the wheat Em gene promoter (Knight et al, 1995) or the soybean GH3 gene promoter (Bierfreund et al, 2003). However, the use of phyto-hormone-regulated gene expression may interfere with the plant physiology and remain unfit for a general versatile conditional gene-expression system in plants.…”

Section: Introductionmentioning

confidence: 99%

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

et al. 2005

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The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using b-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25°C. In contrast, a short non-damaging heat-treatment at 38°C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol (BA) also induced GUS expression at 25°C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens.Abbreviations: ASA, acetyl salicylic acid; BA, benzyl alcohol; GFP, green fluorescent protein; GT, GFPtalin; GUS, b-glucuronidase; Fv/Fm, the maximal photochemical efficiency of PSII; MU, 4-methylumbelliferone; MUG, 4-methylumbelliferyl-b-D-glucuronide; PSII, photosystem II; SA, specific activity; sHSP, small heat-shock protein; TSP, total soluble proteins; Ubi, ubiquitin; X-Gluc, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide

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Use of an inducible reporter gene system for the analysis of auxin distribution in the moss Physcomitrella patens

Cited by 83 publications

References 29 publications

Auxin Metabolism and Function in the Multicellular Brown AlgaEctocarpus siliculosus

Auxin Metabolism and Function in the Multicellular Brown AlgaEctocarpus siliculosus

Homologues of the Arabidopsis thaliana SHI/STY/LRP1 genes control auxin biosynthesis and affect growth and development in the moss Physcomitrella patens

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

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