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Brassinosteroids signal through two receptor-like kinases.

Both animals and plants use steroids to regulate their growth and development, but their mechanisms for steroid perception are different. Animal steroids are mainly recognized by intracellular steroid receptors, whereas plant steroids are perceived by cell-surface receptors that contain a transmembrane receptor serine/threonine kinase. Recent studies suggest that heterodimerization between two receptor kinases might be a key step in steroid perception and signaling in plants.

Brassinosteroids↗

Molecular characterization of the brassinosteroid-deficient lkb mutant in pea.

The brassinosteriod-deficient lkb mutant of garden pea (Pisum sativum L.) is characterized by an erectoides phenotype (reduced internode length, thickened stems, epinastic leaves), which is rescued by application of exogenous brassinolide. We show that the LKB gene is the Arabidopsis DIMINUTO/DWARF-1 (DIM/DWF1) homologue of pea. The DIM/DWF1 homologue from lkb plants contains a mutation that may result in reduced enzyme function, thus resulting in the previously shown accumulation of 24-methylenecholesterol and a deficiency of its hydrogenated product, campesterol. This ultimately leads to a deficiency of the biologically active brassionolide. The mutation in the lkb sequence cosegregates with the lkb phenotype. Northern analyis of the LKB gene revealed that the gene is ubiquitously expressed around the plant and that there is no evidence for negative feedback regulation of the gene.

Amino Acid Sequence↗

Arabidopsis RAV1 is down-regulated by brassinosteroid and may act as a negative regulator during plant development.

RAV1 is a novel DNA-binding protein with two distinct DNA-binding domains unique in higher plants, but its role in plant growth and development remains unknown. Using cDNA array, we found that transcription of RAV1 is down-regulated by epibrassinolide (epiBL) in Arabidopsis suspension cells. RNA gel blot analysis revealed that epiBL-regulated RAV1 transcription involves neither protein phosphorylation/dephosphorylation nor newly synthesized protein, and does not require the functional BRI1, suggesting that this regulation might be through a new BR signaling pathway. Overexpressing RAV1 in Arabidopsis results in a retardation of lateral root and rosette leaf development, and the underexpression causes an earlier flowering phenotype, implying that RAV1 may function as a negative regulatory component of growth and development.

2,4-Dichlorophenoxyacetic Acid↗

Brassinosteroids.

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Brassinosteroids↗

A role for brassinosteroids in the regulation of photosynthesis in Cucumis sativus.

The effects of 24-epibrassinolide (EBR) spray application on gas-exchange, chlorophyll fluorescence characteristics, Rubisco activity, and carbohydrate metabolism were investigated in cucumber (Cucumis sativus L. cv. Jinchun No. 3) plants grown in a greenhouse. EBR significantly increased the light-saturated net CO(2) assimilation rate (A(sat)) from 3 h to 7d after spraying, with 0.1 mg l(-1) EBR proving most effective. Increased A(sat) in EBR-treated leaves was accompanied by increases in the maximum carboxylation rate of Rubisco (V(c,max)) and in the maximum rate of RuBP regeneration (J(max)). EBR-treated leaves also had a higher quantum yield of PSII electron transport (phi(PSII)) than the controls, which was mainly due to a significant increase in the photochemical quenching (q(P)), with no change in the efficiency of energy capture by open PSII reaction centres (F'(v)/F'(m)). EBR did not influence photorespiration. In addition, significant increases in the initial activity of Rubisco and in the sucrose, soluble sugars, and starch contents were observed followed by substantial increases in sucrose phosphate synthase (SPS), sucrose synthase (SS), and acid invertase (AI) activities after EBR treatment. It was concluded that EBR increases the capacity of CO(2) assimilation in the Calvin cycle, which was mainly attributed to an increase in the initial activity of Rubisco.

Brassinosteroids↗

Synthesis and bioactivity of C-29 brassinosteroid analogues with different functional groups at C-6.

In this paper, we report the synthesis and bioactivity of four synthetic analogues of 28-homobrassinosteroids, in order to evaluate the influence in bioactivity when the C-6 keto group is replaced by different functional groups. The synthetic analogues are 6-deoxo-28-homocastasterone [(22R,23R)-stigmasta-2alpha,3alpha,22,23-tetraol], 6alpha-hydroxy-28-homocastasterone [(22R,23R)-stigmasta-2alpha,3alpha,6alpha,22,23-pentaol], 6beta-hydroxy-28-homocastasterone [(22R,23R)-stigmasta-2alpha,3alpha,6beta,22,23-pentaol], and [(22R,23R)-6alpha-fluorostigmasta-2alpha,3alpha,22,23-tetraol]. Results indicate that replacement of the 6-keto moiety by an beta or alpha hydroxyl group led to a decrease in activity, whereas the 6-deoxo analogue showed a very low activity, confirming the importance of an electronegative moiety at C-6 to observe hormonal potency. The 6alpha-fluorinated analogue elicited a low activity, similar to that of the 6-deoxo analogue.

Cholestanones↗

Brassinosteroid-induced exaggerated growth in hydroponically grown Arabidopsis plants.

The effects of root application of brassinolide (BL) on the growth and development of Arabidopsis plants (Arabidopsis thaliana ecotype Columbia [L.] Heynh) were evaluated. Initially, all leaves were evaluated on plants 18, 22, 26 and 29 days old. The younger leaves were found to exhibit maximal petiole elongation and upward leaf bending in response to BL treatment. Therefore, based on these results leaves 6, 7 and 8 on 22-24-day-old plants were selected for all subsequent studies. Elongation along the length of the petiole in response to BL treatment was uniform with the exception of an approximately 4 mm region next to the leaf where upward curvature was observed. Both BL and 24-epibrassinolide (24-epiBL) were evaluated, with BL being more effective at lower concentrations than 24-epiBL. The exaggerated growth induced by 0.1 µM BL was not observed in plants treated with 1 000-fold higher concentrations of GA3, IAA, NAA or 2,4-D (100 µM). In addition, no exaggerated growth effects were observed when plants were treated with 200 ppm ethylene or 1 mM ACC. All treatments with BL, NAA, 2,4-D, IAA or ACC promoted ethylene and ACC production in wild type Arabidopsis plants, but only BL triggered exaggerated plant growth. BL also promoted exaggerated growth and elevated levels of ACC and ethylene in the ethylene insensitive mutant etr1-3, showing that the effect of BR on growth is independent of ethylene. This work provides evidence that BR-induced exaggerated growth of Arabidopsis plants is independent of gibberellins, auxins and ethylene.

Journal Article↗

Nuclear protein phosphatases with Kelch-repeat domains modulate the response to brassinosteroids in Arabidopsis.

Perception of the plant steroid hormone brassinolide (BL) by the membrane-associated receptor kinase BRI1 triggers the dephosphorylation and accumulation in the nucleus of the transcriptional modulators BES1 and BZR1. We identified bsu1-1D as a dominant suppressor of bri1 in A abidopsis. BSU1 encodes a nuclear-localized serine-threonine protein phosphatase with an N-terminal Kelch-repeat domain, and is preferentially expressed in elongating cells. BSU1 is able to modulate the phosphorylation state of BES1, counter acting the action of the glycogen synthase kinase-3 BIN2, and leading to inc eased steady-state levels of dephosphorylated BES1. BSU1 belongs to a small gene family; loss-of-function analyses unravel the extent of functional overlap among members of the family and confirm the role of these phosphatases in the control of cell elongation by BL. Our data indicate that BES1 is subject to antagonistic phosphorylation and dephosphorylation reactions in the nucleus, which fine-tune the amplitude of the response to BL.

Amino Acid Sequence↗

The last reaction producing brassinolide is catalyzed by cytochrome P-450s, CYP85A3 in tomato and CYP85A2 in Arabidopsis.

Brassinosteroids are steroidal hormones essential for the growth and development of plants. Brassinolide, the most biologically active brassinosteroid, has a seven-membered lactone ring that is formed by a Baeyer-Villiger oxidation of its immediate precursor castasterone. Despite its potential key role in controlling plant development, brassinolide synthase has not been identified. Previous work has shown that the formation of castasterone from 6-deoxocastasterone is catalyzed by members of the CYP85A family of cytochrome P-450 monooxygenases. A null mutation in the tomato Dwarf (CYP85A1) gene, extreme dwarf (d(x)), causes severe dwarfism due to brassinosteroid deficiency, but the d(x) mutant still produces fruits. Here, we show that d(x) fruits contain brassinolide at a higher level than wild-type fruits and that a new CYP85A gene, CYP85A3, is preferentially expressed in tomato fruits. Tomato CYP85A3 catalyzed the Baeyer-Villiger oxidation to produce brassinolide from castasterone in yeast, in addition to the conversion of 6-deoxocastasterone to castasterone. We also show that Arabidopsis CYP85A2, which was initially characterized as castasterone synthase, also has brassinolide synthase activity. Exogenous application of castasterone and brassinolide to the Arabidopsis cyp85a1/cyp85a2 double mutant suggests that castasterone can function as an active brassinosteroid but that its conversion into brassinolide is necessary for normal vegetative development in Arabidopsis. We postulate that castasterone is the major active brassinosteroid during vegetative growth in tomato, whereas brassinolide may play an organ-specific role in fruit development in this species.

Arabidopsis↗