• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.139-144
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• 2002

Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

• Journal of Integrative Natural Science
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• v.1 no.1
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• pp.24-31
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• 2008

Brassinosteroids (BRs) are a special class of plant steroid hormones that are essential for normal growth and development. Part of confusion is whether BRs are unique to plants, because they have overlapping physiological roles with other better-studied hormones and with physiological responses caused by light. In systems designed to assay for cytokinins, the effects of BRs vary. We measured hypocotyl length for testing the ability of brassinolide (BL) to rescue double mutant between det2 and the photoreceptor null mutant phytochrome B (phyB). PHYB involved in controlling hypocotyl elongation in increased concentration of BL whereas phyBdet2 double mutant just partially rescue to phyB in white and red light indicated the involvement of BRs in PHYB regulated cell elongation. BRs regulated hypocotyl growth was delayed by BAP, a cytokinin treatment but inhibitory effects of BAPs on hypocotyl growth was slightly recovered by BL. The result indicated that the mode of action of BR and cytokinin is independent or sequential in the downstream light-regulated response control on hypocotyl elongation and also light modulated the action of BR and cytokinin in some extent.

• Molecules and Cells
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• v.39 no.6
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• pp.484-494
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• 2016

Plant cells have a remarkable ability to induce pluripotent cell masses and regenerate whole plant organs under the appropriate culture conditions. Although the in vitro regeneration system is widely applied to manipulate agronomic traits, an understanding of the molecular mechanisms underlying callus formation is starting to emerge. Here, we performed genome-wide transcriptome profiling of wild-type leaves and leaf explant-derived calli for comparison and identified 10,405 differentially expressed genes (> two-fold change). In addition to the well-defined signaling pathways involved in callus formation, we uncovered additional biological processes that may contribute to robust cellular dedifferentiation. Particular emphasis is placed on molecular components involved in leaf development, circadian clock, stress and hormone signaling, carbohydrate metabolism, and chromatin organization. Genetic and pharmacological analyses further supported that homeostasis of clock activity and stress signaling is crucial for proper callus induction. In addition, gibberellic acid (GA) and brassinosteroid (BR) signaling also participates in intricate cellular reprogramming. Collectively, our findings indicate that multiple signaling pathways are intertwined to allow reversible transition of cellular differentiation and dedifferentiation.

• Korean Journal of Soil Science and Fertilizer
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• v.32 no.1
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• pp.57-61
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• 1999

This study measured the soaking effects of two brassinosteroids(BRs), homobrassinolide(hBR : 225, 23S-homobrassinolide) and epibrassinolide(epiBR : 22R, 23R. 24R-$2{\alpha}$, $3{\alpha}$, 22, 23-Tetrahydroxy-B-homo-7-oxa-$5{\alpha}$-ergostan-6-one) on the germination of two rice cultivars, 'Milyang 23' and 'Naepungbyeo' exposed to simulated acid rain(SAR). The results suggested that promoting effect of two BRs on alleviating damage to SAR would be attributed to the promotion of plant growth, root elongation, and particularly germination rate of both cultivars. Eight organic acids were determined by HPLC. Succinic acid acetic acid in 'Milyang 23' under SAR were highest among eight organic acids.

• Journal of Life Science
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• v.21 no.5
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• pp.706-712
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• 2011

Metabolism of $^3H$-castasterone in the presence of $^{32}P$-ATP was examined by an enzyme solution prepared from A. thaliana after a reversed phased HPLC, after which a polar metabolite labeled by both $^3H$ and $^{32}P$ was obtained, suggesting that $^3H$-CS is phosphorylated by $^{32}P$-ATP. To elucidate the structure of the phosphorylated CS, the same enzyme assay was carried out with non-isotopes labeled CS and ATP. In GC-MS analysis the metabolite gave a molecular ion at m/z 664 as a bismethanboronate, suggesting the metabolite is a CS phosphate. Treatment of wheat germ acid phosphatase that hydrolyzed phosphoester bond gave the same mass spectrum and GC retention time in GC-MS analyses, confirming that the metabolite is phosphorylated CS. This is the first example of phosphorylated conjugates of CS in plants.

• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.30-36
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• 2016

Brassinosteroids (BRs) are essential plant steroid hormones required for cell elongation, plant growth, development and abiotic and biotic stress tolerance. BRs are recognized by BRI1 receptor kinase that is localized in the plasma membrane, and the BRI1 protein will eventually autophosphorylate in the intracellular domain and transphosphorylate BAK1, which is a co-receptor in Arabidopsis thaliana. However, little is known of the role OsBRI1 receptor kinase plays in Oryza sativa, monocotyledonous plants, compared to that in Arabidopsis thaliana, dicotyledonous plants. As such, we have studied OsBRI1 receptor kinase in vitro and in vivo with recombinant protein and transgenic plants, whose phenotypes were also investigated. A OsBRI1 cytoplasmic domain (CD) recombinant protein was induced in BL21 (DE3) E.coli cells with IPTG, and purified to obtain OsBRI1 recombinant protein. Based on Western blot analysis with phospho-specific pTyr and pThr antibodies, OsBRI1 recombinant protein and OsBRI1-Flag protein were phosphorylated on Threonine residue(s), however, not on Tyrosine residue(s), both in vitro and in vivo. This is particularly intriguing as AtBRI1 protein was phosphorylated on both Ser/Thr and Tyr residues. Also, the OsBRI1 full-length gene was expressed in, and rescued, bri1-5 mutants, such as is seen in normal wild-type plants where AtBRI1-Flag rescues bri1-5 mutant plants. Root growth in seedlings decreased in Ws2, AtBRI1, and 3 independent OsBRI1 transgenic seedlings and had an almost complete lack of response to brassinolide in the bri1-5 mutant. In conclusion, OsBRI1, an orthologous gene of AtBRI1, can mediate normal BR signaling for plant growth and development in Arabidopsis thaliana.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.161-167
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• 2005

Brassinosteroids are steroidal plant hormones and are known to modulate physiological and cellular response in a wide range of plant species. Considerable insights has been achieved of the physiological role of brassinosteroid in Brassica species in the past few years, but their effect on direct organogenesis has not been extensively studied. In this sense, under optimal basal media and growth conditions we tested the cellular and organogenic response of 24-epibrassinolide (EBL) in a variable concentration (0.1 to $5.0\;{\mu}M$) and Zeatin (Z) (1.0 to $100\;{\mu}M$) and their synergic effect on hypocotyl explants of cauliflower and broccoli. The isolated EBL accelerated cell elongation and promotes direct organogenesis. One micromolar EBL + $10\;{\mu}M$ of Z was the most efficient combination for cell elongation, cell differentiation as well as for organogenesis. A suppressing effect on root induction was confirmed for all the tested hormone levels. The general results indicate a synergic effect of EBL-Z and EBL potentates Zeatin activity, at least in certain tissues. Besides de genetic factors, we can speculate that the natural hormone concentration in the explants might affect the responses by application of exogenous growth regulators. Experiments with new plant growth regulators, like brassinolide, are important aiming to maximize or accelerate plant regeneration for in vitro multiplication or for genetic transformation.

• Molecules and Cells
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• v.27 no.2
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• pp.183-190
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• 2009

The plasma membrane-localized BRASSINOSTEROID-INSENSITIVE1 (BRI1) and BRI1-ASSOCIATED KINASE1 (BAK1) are a well-known receptor pair involved in brassinosteroids (BR) signaling in Arabidposis. The formation of a receptor complex in response to BRs and the subsequent activation of cytoplasmic domain kinase activity share mechanistic characteristics with animal receptor kinases. Here, we demonstrate that BRI1 and BAK1 are BR-dependently phosphorylated, and that phosphorylated forms of the two proteins persist for different lengths of time. Mutations of either protein abolished phosphorylation of the counterpart protein, implying transphosphorylation of the receptor kinases. To investigate the specific amino acids critical for formation of the receptor complex and activation of BAK1 kinase activity, we expressed several versions of BAK1 in yeast and plants. L32E and L46E substitutions resulted in a loss of binding of BAK1 to BRI1, and threonine T455 was essential for the kinase activity of BAK1 in yeast. Transgenic bri1 mutant plants overexpressing BAK1(L46E) displayed reduced apical dominance and seed development. In addition, transgenic wild type plants overexpressing BAK1(T455A) lost the phosphorylation activity normally exhibited in response to BL, leading to semi-dwarfism. These results suggest that BAK1 is a critical component regulating the duration of BR efficacy, even though it cannot directly bind BRs in plants.

• Molecules and Cells
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• v.37 no.11
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• pp.795-803
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• 2014

To withstand ever-changing environmental stresses, plants are equipped with phytohormone-mediated stress resistance mechanisms. Salt stress triggers abscisic acid (ABA) signaling, which enhances stress tolerance at the expense of growth. ABA is thought to inhibit the action of growth-promoting hormones, including brassinosteroids (BRs). However, the regulatory mechanisms that coordinate ABA and BR activity remain to be discovered. We noticed that ABA-treated seedlings exhibited small, round leaves and short roots, a phenotype that is characteristic of the BR signaling mutant, brassinosteroid insensitive1-9 (bri1-9). To identify genes that are antagonistically regulated by ABA and BRs, we examined published Arabidopsis microarray data sets. Of the list of genes identified, those upregulated by ABA but downregulated by BRs were enriched with a BRRE motif in their promoter sequences. After validating the microarray data using quantitative RT-PCR, we focused on RD26, which is induced by salt stress. Histochemical analysis of transgenic Arabidopsis plants expressing RD26pro:GUS revealed that the induction of GUS expression after NaCl treatment was suppressed by co-treatment with BRs, but enhanced by co-treatment with propiconazole, a BR biosynthetic inhibitor. Similarly, treatment with bikinin, an inhibitor of BIN2 kinase, not only inhibited RD26 expression, but also reduced the survival rate of the plant following exposure to salt stress. Our results suggest that ABA and BRs act antagonistically on their target genes at or after the BIN2 step in BR signaling pathways, and suggest a mechanism by which plants fine-tune their growth, particularly when stress responses and growth compete for resources.

• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.280-286
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• 2007

We evaluated a commercial biopreparation of plant growth-promoting rhizobacteria (PGPR) strains Bacillus subtilis GB03 and B. amyloliquefaciens IN937a formulated with the carrier chitosan (Bio Yield) for its capacity to elicit growth promotion and induced systemic resistance against infection by Cucumber Mosaic Virus (CMV) and Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana. The biopreparation promoted plant growth of Arabidopsis hormonal mutants, which included auxin, gibberellic acid, ethylene, jasmonate, salicylic acid, and brassinosteroid insensitive lines as well as each wild-type. The biopreparation protected plants against CMV based on disease severity in wild-type plants. However, virus titre was not lower in control plants and those treated with biopreparation, suggesting that the biopreparation induced tolerance rather than resistance against CMV. Interestingly, the biopreparation induced resistance against CMV in NahG plants, as evidenced by both reduced disease severity and virus titer. The biopreparation also elicited induced resistance against P. syringae pv. tomato in the wild-type but not in NahG transgenic plants, which degrade endogenous salicylic acid, indicating the involvement of salicylic acid signaling. Our results indicate that some PGPR strains can elicit plant growth promotion by mechanisms that are different from known hormonal signaling pathways. In addition, the mechanism for elicitation of induced resistance by PGPR may be pathogen-dependent. Collectively, the two-Bacilli strain mixture can be utilized as a biological inoculant for both protection of plant against bacterial and viral pathogens and enhancement of plant growth.