• 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 Integrative Natural Science
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• v.4 no.2
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• pp.113-120
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• 2011

Brassinosteroids (BRs) are plant steroid hormones that play essential roles in growth and development. Mutations in BR-signaling pathways cause defective in growth and development like dwarfism, male sterility, abnormal vascular development and photomorphogenesis. Transition from vegetative to reproductive growth is a critical phase change in the development of a flowering plant. In a screen of activation-tagged Arabidopsis, we identified a mutant named abz126 that displayed longer hypocotyls when grown in the dark on MS media containing brassinazole (Brz), an inhibitor of BRs biosynthesis. We have cloned the mutant locus using adapter ligation PCR walking and identified that a single T-DNA had been integrated into the ninth exon of the GIGANTEA (GI) gene, involved in controling flowering time. This insertion resulted in loss-of-function of the GI gene and caused the following phenotypes: long petioles, tall plant height, many rosette leaves and late flowering. RT-PCR assays on abz126 mutant showed that the T-DNA insertion in GIGANTEA led to the loss of mRNA expression of the GI gene. In the hormone dose response assay, abz126 mutant showed: 1) an insensitivity to paclobutrazole (PAC), 2) an altered response with 6-benzylaminopurine (BAP) and 3) insensitive to Brassinolide (BL). Based on these results, we propose that the late flowering and tall phenotypes displayed by the abz126 mutant are caused by a loss-of-function of the GI gene associated with brassinosteroid hormone signaling.

• Korean Journal of Plant Tissue Culture
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• v.21 no.5
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• pp.281-286
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• 1994

The hormonal regulation of organ differentiation was investigated in the tissue culture of sweet potato. Embryogenic callus was induced from shoot tips cultured on MS medium supplemented with 1 mg/L 2,4-D. When embryogenic callus was transferred to medium containing 0.1 mg/L GA$_4$, it proliferation was stimulated. The callus gave rise to plantlets when cultured on medium containing 0.1 mg/L BA. Addition of 0.1 mg/L jasmonic acid or 0.01 mg/L brassinolide to medium was effective for the development of healthy normal plantlets.

• Molecules and Cells
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• v.41 no.12
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• pp.1072-1080
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• 2018

A plant-specific B3 domain and AP2 domain-containing transcription factor, RAV1 acts as a negative regulator of growth in many plant species and its transcription was down-regulated by BR and ABA. In this study, we found that RAV1-overexpressing transgenic plants showed abnormally developed ovules, resulting in reduced seed size, weight, and number in a silique. Interestingly, the endogenous expression of RAV1 fluctuated during seed development; it remained low during the early stage of seed development and sharply increased in the seed maturation stage. In plants, seed development is a complex process that requires coordinated growth of the embryo, endosperm, and maternal integuments. Among many genes that are associated with endosperm proliferation and embryo development, three genes consisting of SHB1, MINI3, and IKU2 form a small unit positively regulating this process, and their expression was regulated by BR and ABA. Using the floral stage-specific RNAs, we found that the expression of MINI3 and IKU2, the two downstream genes of the SHB1-MINI3-IKU2 cascade in the seed development pathway, were particularly reduced in the RAV1-overexpressing transgenic plants. We further determined that RAV1 directly binds to the promoter of MINI3 and IKU2, resulting in their repression. Direct treatment with brassinolide (BL) improved seed development of RAV1-overexpressing plants, but treatment with ABA severely worsened it. Overall, these results suggest that RAV1 is an additional negative player in the early stages of seed development, during which ABA and BR signaling are coordinated.

• Journal of Life Science
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• v.19 no.8
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• pp.1139-1144
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• 2009

It has been known that brassiolide (BL) increased the positive gravitropic response and ethylene production in maize roots. This study examined the relationship between the BL-induced gravitropic response and ethylene Production. The ethylene production was inhibited to about 90% of the control by the treatment of $10^{-4}$ M aminoethoxyvinylglycine (AVG), the ethylene synthesis inhibitor. However, the gravitropic response did not show any significant changes compared to the control at $10^{-4}$ M AVG. In the case of treatment of AVG with BL, the ethylene production decreased to 60% of the control. However, the gravitropic response increased to the level which was induced by BL. Cobalt ions, another ethylene biosynthesis inhibitor, inhibited ethylene production, but not gravitropic response. When roots were treated with BL and cobalt ions, they showed the inhibition of ethylene production and promotion of gravitropic response. To elucidate the possibility that the effect of BL is related to auxin transport, roots were treated with TIBA (2,3,5-triiodobenzoic acid), an auxin transport inhibitor. Both treatment of TIBA alone and TIBA with BL stimulated ethylene production to about 96% and 132%, respectively. However, gravitropic response was completely inhibited in both treatments. Further, roots treated with BL in the presence of TIBA and IAA showed a negative gravitropic response, which means that IAA accumulates in the upper side of horizontal roots. Root elongation was also stimulated in this treatment. Taken together, these results suggest that BL might affect the differential distribution of internal IAA on roots, causing the regulation of positive gravitropic response.