Brassinosteroid biosynthesis and signalling in Petunia hybrida.
A PCR-based approach was performed to identify potential petunia homologues. Due to their appearance, these mutants were named compact disc cd. Such a scenario, where an OGT competes with a kinase, is a widely recognized mechanism in animal research Kamemura et al. To identify proteins that interact with PhBEH2, a yeast two-hybrid screen was performed. In all mutants, the endogenous BR levels were severely reduced. Interestingly, PhBEH2 also interacted with proteins implicated in other signalling pathways.
Journal of Biological Chemistry , — The chemical characteristic and distribution of brassinosteroids in plants. In order to study BR biosynthesis and signalling in petunia, populations of the mutagenic Petunia hybrida line W were visually screened Gerats et al. Developmental Cell 13 , — Only fragmentary details about BR signalling in other plant species are known. C Classification of the positive clones from B.
Contributor Oxford University Press. This article is from Journal of Experimental Botany , volume Abstract Brassinosteroids BRs are steroidal plant hormones that play an important role in the growth and development of plants.
The biosynthesis of sterols and BRs as well as the signalling cascade they induce in plants have been elucidated largely through metabolic studies and the analysis of mutants in Arabidopsis and rice.
Only fragmentary details about BR signalling in other plant species are known. Here a forward genetics strategy was used in Petunia hybrida, by which 19 families with phenotypic alterations typical for BR deficiency mutants were identified. In all mutants, the endogenous BR levels were severely reduced.
Interestingly, PhBEH2 also interacted with proteins implicated in other signalling pathways. This suggests that PhBEH2 might function as an important hub in the cross-talk between diverse signalling pathways. Issn Print Electronic. The underlined sequence indicates the target site duplication. To explore the BR signalling cascade in petunia further, the aim was to identify signalling components downstream of PhBRI1.
A PCR-based approach was performed to identify potential petunia homologues. S2 at JXB online. To identify proteins that interact with PhBEH2, a yeast two-hybrid screen was performed.
As many transcription factors carry activation domains, auto-activation of the full-length cDNA of PhBEH2 on yeast reporters was tested first. Sequence analysis showed that five of these proteins were proteins.
Characterization of PhBEH2 and the identification of binding partners by a yeast two-hybrid screen. C Classification of the positive clones from B. As indicated, for some proteins, a homologue with known function is found in Arabidopsis. All constructs are driven by the 35S promoter.
All panels show a transformed as well as an untransformed protoplast. To verify some of the interactions found in yeast in plant cells, BiFC was used in petunia leaf protoplasts Hu et al. As numerous interactions with PhBEH2 were detected, the search was restricted to interactions known from Arabidopsis research. S3 at JXB online.
This clone was renamed PSK9. To gain more insight into the function of PSK8, a yeast two-hybrid screen was performed, where full-length PSK8 was used as bait to screen a cDNA library made from young petunia inflorescences. MPK1 from Arabidopsis was used as an outgroup to construct the tree, but is not shown in this tree.
B Interaction of PSK8 with eight different clones in a yeast two-hybrid assay. When a clear homologue in Arabidopsis is found, their AGI identifier is listed.
The synthesis of BRs and the signalling cascade downstream of the bioactive compounds have been largely elucidated over the past 10 years Fujioka and Yokota, ; Clouse, As in many plant research areas, these findings basically originate from Arabidopsis.
Partial confirmation of evolutionary conservation as well as additions came from numerous reports on BR biosynthesis and signalling in other species e. The available resources in petunia have been used to identify large parts of the BR biosynthesis and signalling route. The initial results point to conserved BR biosynthesis genes and interacting signalling proteins, as well as to potential new relationships between hormone signalling pathways.
Generally, a mutation in a BR biosynthesis gene causes the accumulation of a precursor s that should have been metabolized and a reduction of the end product s. In the cd2 mutant, accumulation of OH-CR, which has been recognized as the first metabolite of campesterol in the early C oxidation branch of BR biosynthesis, was observed Fujita et al.
Once more the biochemical data were confirmed as cd3 was found to be mutated in the petunia homologue of CYP85A1 Fig. Endogenous levels of upstream BRs in cd1 are quite similar to those of the wild type, and sterol levels are normal.
However, the levels of both 6-deoxoCS and CS, which appear in the last steps of biosynthesis, are reduced. No accumulation of other measured BR biosynthesis intermediates could be identified.
Thus, the nature of the cd1 mutation is currently unknown and it might represent an enzyme that has not been identified yet in any other plant species. Altogether, the evidence available points to unknown BR biosynthesis enzymes or perhaps regulators of BR biosynthesis enzymes mutated in cd1 or cd9.
Cloning of the responsible genes can be accomplished in the future by using amplified fragment length polymorphim AFLP -based transposon display or high throughput sequencing of dTph1 -flanking sequences Van den Broeck et al.
The single signalling mutant that was identified, cd10 , was shown to harbour an insertion in the petunia homologue of the BR receptor, BRI1 Li and Chory et al. The cd10 mutant exhibits an extremely dwarfed stature and did not flower at all.
All known bri1 mutants do flower, except the osbri1 mutant from rice Nakamura et al. In an amino acid alignment of BRI1 from different species, the highest similarity was observed in the crucial kinase domain Supplementary Fig.
S1 at JXB online; Tang et al. Thus it seems that the perception of the BR hormone is evolutionarily well conserved. The present yeast two-hybrid screen with PhBEH2 revealed several interacting proteins that function in diverse signal transduction pathways.
Recently, a number of reports highlighted interactions between BR signalling and other hormone pathways reviewed by Choudhary et al. O -GlcNAcylation of target proteins might affect their activity, localization, stability, and interactions with other proteins.
Such a scenario, where an OGT competes with a kinase, is a widely recognized mechanism in animal research Kamemura et al. Multiple links between petunia BR signalling components and other pathways were identified. One of the RRM proteins is a homologue of FPA , a gene that regulates flowering time in Arabidopsis via a pathway that is independent of daylength Schomburg et al.
Various connections between flowering and BRs have been revealed Domagalska et al. This is consistent with the finding that BR biosynthesis mutants as well as signalling mutants are late flowering or, like the cd10 petunia bri1 mutant, non-flowering. Although interesting, further research should confirm these findings.
Remarkably, no known homologues of the BR signalling pathway were found in the yeast two-hybrid screen performed with PSK8. PSK8 interacted with a number of proteins that are poorly characterized. Here, the identified interactions infer a direct role for GSK3-like proteins in the regulation of enzyme-like proteins, a mechanism that has interfaces with some of the roles of GSK3 in animals.
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Close mobile search navigation Article navigation. Brassinosteroid biosynthesis and signalling in Petunia hybrida Nathalie Verhoef.
Abstract Brassinosteroids BRs are steroidal plant hormones that play an important role in the growth and development of plants. View large Download slide. Overview of cd alleles and results of complementation experiment by brassinolide treatment.
FW, fresh weight; ND, not detectable; lost, internal standards not recovered. Functions of OsBZR1 and proteins in brassinosteroid signaling in rice.
The chemical characteristic and distribution of brassinosteroids in plants. Effects of brassinosteroids on the plant responses to environmental stresses.
The tomato Dwarf gene isolated by heterologous transposon tagging encodes the first member of a new cytochrome P family. Arabidopsis mutants reveal multiple roles for sterols in plant development.
Attenuation of brassinosteroid signaling enhances FLC expression and delays flowering. An early C oxidation branch in the brassinosteroid biosynthetic pathway. An essential role for proteins in brassinosteroid signal transduction in Arabidopsis. Molecular characterization of a nonautonomous transposable element dTph1 of petunia. Brassinolide, a plant growth-promoting steroid isolated from Brassica napus pollen. Two O-linked N-acetylglucosamine transferase genes of Arabidopsis thaliana L.
Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast.
Dynamic interplay between O-glycosylation and O-phosphorylation of nucleocytoplasmic proteins: Brassinosteroid signal transduction from cell-surface receptor kinases to nuclear transcription factors. Binding of brassinosteroids to the extracellular domain of plant receptor kinase BRI1.
A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction. Accumulation of 6-deoxocathasterone and 6-deoxocastasterone in Arabidopsis, pea and tomato is suggestive of common rate-limiting steps in brassinosteroid biosynthesis. PH4 of Petunia is an R2R3 MYB protein that activates vacuolar acidification through interactions with basic-helix—loop—helix transcription factors of the anthocyanin pathway.
Nucleocytoplasmic shuttling of BZR1 mediated by phosphorylation is essential in Arabidopsis brassinosteroid signaling. Brassinosteroidoxidases from Arabidopsis and tomato catalyze multiple C-6 oxidations in brassinosteroid biosynthesis. Brassinosteroids rescue the deficiency of CYP90, a cytochrome P, controlling cell elongation and de-etiolation in Arabidopsis.
Transposon display identifies individual transposable elements in high copy number lines. Generation of a 3D indexed Petunia insertion database for reverse genetics. Analysis of flower pigmentation mutants generated by random transposon mutagenesis in Petunia hybrida. Cross-talk between GlcNAcylation and phosphorylation: Dynamic interplay between O-linked N-acetylglucosaminylation and glycogen synthase kinasedependent phosphorylation. Nuclear-localized BZR1 mediates brassinosteroid-induced growth and feedback suppression of brassinosteroid biosynthesis.
BES1 accumulates in the nucleus in response to brassinosteroids to regulate gene expression and promote stem elongation. Two putative BIN2 substrates are nuclear components of brassinosteroid signaling. For commercial re-use, please contact journals. Email alerts New issue alert. Receive exclusive offers and updates from Oxford Academic. Related articles in Web of Science Google Scholar.
Citing articles via Web of Science 4. Selection of Host-Plant Genotype.
Iamges: brassinosteroid biosynthesis and signalling in petunia hybrida
In plants carrying the cd3 D allele, a transposable element could not be identified in the petunia CYP85A1 homologue.
Sequence analysis showed that five of these proteins were proteins. Gene Ontology GO Terms.
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