• Journal of Plant Biotechnology
• /
• 제43권1호
• /
• pp.30-36
• /
• 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.

• 통합자연과학논문집
• /
• 제4권2호
• /
• pp.113-120
• /
• 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.

• 식물조직배양학회지
• /
• 제21권5호
• /
• pp.281-286
• /
• 1994

고구마 경단배양에 의한 우량종묘 대량증식 과정에서 분화 및 생육에 미치는 생장조절제의 효과를 구명하기 위하여 실험을 수행하였다. 고구마 경단조직은 1 mEm/L 2,4-D 첨가된 MS배지에서 배양 40일경 배발생 캘러스가 형성되었다. 또한 형성된 배발생 캘러스는 0.1 mg/L GA$_4$ 첨가된 배지에 배양한 경우, 효과적으로 배발생 캘러스를 유지 및 증식이 가능하였으며, 배발생캘러스로부터 shoot 재분화는 0.1mg/L BA가 유효하였다. 체세포배로부터 shoot 재분화는 0.01 ngh BA와 0.01 mg/L GA$_4$ 혹은 0.1 mg/L BA와 0.01mg/L GA$_4$ 혼합배지에서 양호하였으며, 0.1-1 mg/L BA를 첨가된 배지에서는 배발생 캘러스 및 체세포배로부터 녹색의 단단한 캘러스가 형성되었다. 체세포배를 배양하여 얻어진 5 mm 크기의 shoot 정단을 재배양 시, 0.1 mg/L jasmonic acid 또는 0.01 mg/L brassnlolide가 첨가된 배지에서 소식물체의 생육이 가장 양호하였다.

• Molecules and Cells
• /
• 제41권12호
• /
• pp.1072-1080
• /
• 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.

• 생명과학회지
• /
• 제19권8호
• /
• pp.1139-1144
• /
• 2009

옥수수 뿌리에 BL을 처리하면 양성굴중성 반응이 촉진되고, ethylene 생성도 증가한다는 것이 알려져 있다. BL에 의해 유도된 굴중성 반응과 ethylene 생성과의 관계를 조사하였다. ethylene 생성 억제제인 $10^{-4}$ M AVG를 처리하면 ethylene 생성은 90% 이상 억제되었으나, 굴중성 반응은 13% 정도 억제되었다. AVG를 BL과 함께 처리한 뿌리는 ethylene 생성은 약 60% 억제되었으나 굴중성 반응은 대조구 보다 증가하였다. 다른 ethylene 생성 억제제인 cobalt ion을 처리하면 ethylene 생성은 약 10% 정도 억제되었으나 굴중성 반응은 억제되지 않았다. BL과 cobalt ion을 함께 처리한 뿌리는 ethylene 생성이 억제되었으나 굴중성 반응은 증가되었다. 이러한 BL의 효과가 auxin transport와 관계가 있는지 알아보기 위하여 auxin transport inhibitor인 TIBA를 처리하였다. $10^{-5}$ M TIBA와 BL과 TIBA를 함께 처리한 경우(BL+TIBA), ethylene 생성은 각각 96%, 132% 증가하였으나 굴중성 반응은 모두 일어나지 않았다. 또한, BL, TIBA 그리고 IAA를 함께 처리한 뿌리 (BL+TIBA+IAA)는 음성굴중성 반응을 나타냈으나 뿌리 생장은 오히려 증가시켰으며, 이는 수평으로 있는 뿌리에서 IAA가 아랫면으로 transport 되지 못하고 윗면에 축적된 것을 의미한다. 이러한 결과는 BL이 뿌리 내에 존재하는 IAA의 차등분포에 영향을 주어 양성굴중성 반응을 촉진할 가능성을 제시한다.