Dataset Information

Brassinosteroids Induce Strong, Dose-Dependent Inhibition of Etiolated Pea Seedling Growth Correlated with Ethylene Production.

ABSTRACT: We have recently discovered that brassinosteroids (BRs) can inhibit the growth of etiolated pea seedlings dose-dependently in a similar manner to the 'triple response' induced by ethylene. We demonstrate here that the growth inhibition of etiolated pea shoots strongly correlates with increases in ethylene production, which also responds dose-dependently to applied BRs. We assessed the biological activities of two natural BRs on pea seedlings, which are excellent material as they grow rapidly, and respond both linearly and uni-phasically to applied BRs. We then compared the BRs' inhibitory effects on growth, and induction of ethylene and ACC (1-aminocyclopropane-1-carboxylic acid) production, to those of representatives of other phytohormone classes (cytokinins, auxins, and gibberellins). Auxin induced ca. 50-fold weaker responses in etiolated pea seedlings than brassinolide, and the other phytohormones induced much weaker (or opposite) responses. Following the optimization of conditions for determining ethylene production after BR treatment, we found a positive correlation between BR bioactivity and ethylene production. Finally, we optimized conditions for pea growth responses and developed a new, highly sensitive, and convenient bioassay for BR activity.

SUBMITTER: Jiroutova P

PROVIDER: S-EPMC7017382 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Brassinosteroids Induce Strong, Dose-Dependent Inhibition of Etiolated Pea Seedling Growth Correlated with Ethylene Production.

Jiroutová Petra P Mikulík Jaromír J Novák Ondřej O Strnad Miroslav M Oklestkova Jana J

Biomolecules 20191209 12

We have recently discovered that brassinosteroids (BRs) can inhibit the growth of etiolated pea seedlings dose-dependently in a similar manner to the 'triple response' induced by ethylene. We demonstrate here that the growth inhibition of etiolated pea shoots strongly correlates with increases in ethylene production, which also responds dose-dependently to applied BRs. We assessed the biological activities of two natural BRs on pea seedlings, which are excellent material as they grow rapidly, an ...[more]

PMID: 31835421

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Project description:To better understand light regulation of C(4) plant maize development, we investigated dynamic proteomic differences between green seedlings (control), etiolated seedlings, and etiolated seedlings illuminated for 6 or 12 h using a label-free quantitative proteomics approach based on nanoscale ultraperformance liquid chromatography-ESI-MS(E). Among more than 400 proteins identified, 73 were significantly altered during etiolated maize seedling greening. Of these 73 proteins, 25 were identified as membrane proteins that seldom had been identified with two-dimensional electrophoresis methods, indicating the power of our label-free method for membrane protein identification; 31 were related to light reactions of chlorophyll biosynthesis, photosynthesis, and photosynthetic carbon assimilation. The expression of photosystem II subunits was highly sensitive to light; most of them were not identified in etiolated maize seedlings but drastically increased upon light exposure, indicating that the complex process of biogenesis of the photosynthetic apparatus correlates with the transition from a dark-grown to a light-grown morphology. However, transcriptional analysis indicated that most transcripts encoding these proteins were not regulated by light. In contrast, the levels of mRNAs and proteins for enzymes involved in carbon assimilation were tightly regulated by light. Additionally phosphoenolpyruvate carboxykinase, the key enzyme of the phosphoenolpyruvate carboxykinase C(4) pathway, was more tightly regulated by light than the key enzymes of the NADP-malic enzyme C(4) pathway. Furthermore phosphoenolpyruvate carboxylase 1C, which was originally reported to be specifically expressed in roots, was also identified in this study; expression of this enzyme was more sensitive to light than its isoforms. Taken together, these results represent a comprehensive dynamic protein profile and light-regulated network of C(4) plants for etiolated seedling greening and provide a basis for further study of the mechanism of gene function and regulation in light-induced development of C(4) plants.

Project description:The proper transition between the skotomorphogenic and photomorphogenic developmental programs is key for seedling survival and it is therefore tightly regulated. Besides light, which is the major cue that triggers this transition, several hormone pathways participate in this control, mainly antagonizing the light effect. Two of these hormones are gibberellins (GA) and brassinosteroids (BR). Both hormones promote the skotomorphogenesis and prevent photomorphogenesis, as evidenced by the partially de-etiolated phenotype caused by GA or BR deficiency, or by a blockage in the respective signaling pathways. We have previously shown that BR mediate GA activity in the control of this transition. For instance, treatment of dark-grown, GA-deficient seedlings with BRs was able to partially restore morphological phenotypes such as hypocotyl growth, and importantly to restore completely the molecular phenotype of CAB2 and RbcS gene expression. On the contrary, GA-treatment did not alter the same phenotypes in the BR deficient det2 mutant. Thus, the aim of this study was to investigate to what extent BRs mediate GA activity in dark-grown seedlings, and for that purpose we have the examined global changes in gene expression caused by treatment with GA and BRs in BR- and GA-deficient seedlings, respectively. We performed two sets of experiments, each one including three different samples. The first experiment included dark-grown, WT Col-0 mock-treated seedlings, or seedlings treated with either the GA biosynthesis inhibitor paclobutrazol (PAC) or with PAC+epibrassinolide (EBR). In this experiment, PAC samples were labelled with Cy3 and compared to WT and PAC+EBR samples, which were labelled with Cy5. Two biological repeast were performed. The second experiment included dark-grown, WT Col-0 seedlings and the det2-1 mutant, which was either mock-treated or treated with GA3. Two biological replicates were performed, in the first one the WT and det2-1+GA3 were labelled with Cy3 and the det2-1 sample with Cy5. The reverse labelling was used in the second replicate.

Dataset Information

Brassinosteroids Induce Strong, Dose-Dependent Inhibition of Etiolated Pea Seedling Growth Correlated with Ethylene Production.

Publications

Brassinosteroids Induce Strong, Dose-Dependent Inhibition of Etiolated Pea Seedling Growth Correlated with Ethylene Production.

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