• International Journal of Industrial Entomology and Biomaterials
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• 제11권1호
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• pp.67-70
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• 2005

A cuticle protein gene, PbLCP12.1, from the white­spotted flower chafer, Protaetia brevitarsis, was isolated and characterized. The gene contains an ORF of 336 nucleotides capable of encoding a 113 amino acid polypeptide with a predicted molecular mass of 12,138 Da and pI of 4.15. The PbLCP12.1 protein contained a type-specific consensus sequence identifiable in other insect cuticle proteins. The deduced amino acid sequence of the PbLCP12.1 cDNA is most similar to Bombyx mori cuticle protein BmLCP18 (37$\%$ protein sequence identity). Northern blot analysis revealed that PbLCP12.1 showed the epidermis-specific expression.

• Journal of Plant Biotechnology
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• 제45권4호
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• 원예과학기술지
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• 제29권6호
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• pp.511-522
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• 2011

화색은 화훼 육종의 주요한 목표형질이다. 최근 유전공학 기술의 발달로 기존의 전통육종에서는 볼 수 없었던 파란장미와 파란카네이션과 같은 새로운 화색 개발이 성공적으로 보고되었다. 플라보노이드 생합성에 관해 축적된 지식기반 연구 결과를 바탕으로 새롭고 독특한 형질의 화색을 도입하는 것이 가능하게 된 것이다. 이러한 화색변경은 플라보노이드 대사경로의 조절, 즉 내재유전자의 발현조절 및 새로운 플라보노이드 합성 또는 특정 플라보노이드 합성을 위한 외래유전자의 추가도입과 플라보노이드 대사 전체를 조절하는 전사인자의 도입을 통해서 이루어져 왔다. 그러나 보다 실증적으로 이러한 플라보노이드 대사를 조절하기 위해서는 작물별 내재 플라보노이드의 조절 기작에 대한 이해를 바탕으로 목표로 하는 플라보노이드 합성을 위해 보다 정교한 대사흐름의 조절이 요구된다. 본 총설에서는 화훼작물의 화색변경 성공 예들을 자세히 소개하고 그 요인 분석을 통해 향후 더 성공적인 화색변경의 전략을 수립하는데 도움이 되고자 한다.

• Molecules and Cells
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• 제25권2호
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• pp.294-300
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• 2008

Cytokinins are essential hormones in plant development. $\underline{A}$rabidopsis $\underline{h}$istidine-containing $\underline{p}$hosphotransfer proteins (AHPs) are mediators in a multistep phosphorelay pathway for cytokinin signaling. The exact role of AHP4 has not been elucidated. In this study, we demonstrated young flower-specific expression of AHP4, and compared AHP4-overexpressing (Ox) trangenic Arabidopsis lines and an ahp4 knock-out line. AHP4-Ox plants had reduced fertility due to a lack of secondary cell wall thickening in the anther endothecium and inhibition of IRREGURAR XYLEMs (IRXs) expression in young flowers. Conversely, ahp4 anthers had more lignified anther walls than the wild type, and increased IRXs expression. Our study indicates that AHP4 negatively regulates thickening of the secondary cell wall of the anther endothecium, and provides new insight into the role of cytokinins in formation of secondary cell walls via the action of AHP4.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• 한국식품저장유통학회지
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• 제22권5호
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• pp.744-750
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• 2015

본 연구에서는 연화 열수 및 에탄올 추출물의 3T3-L1 지방세포 분화 및 ROS생성 저감효과를 구명하기 위하여 3T3-L1 전지방세포 분화과정 중 연화 열수 및 에탄올 추출물에 의한 지방축적과 ROS 생성을 관찰하였다. 연화 열수 및 에탄올 추출물은 XTT assay에서 100, 200 및 $400{\mu}g/mL$ 농도에서 세포 독성을 보이지 않았다. 지방세포 분화 중 세포 내 지방축적 및 ROS 생성량을 비교한 결과, 연화 열수 및 에탄올 추출물을 처리한 지방세포의 경우 지방축적량과 ROS 생성량 모두 유의적으로 억제되는 것으로 나타났다. 특히 연화 열수 및 에탄올 추출물을 처리함으로써 지방세포분화와 관련된 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$ 및 aP mRNA의 발현을 유의적으로 감소시켰으며, ROS의 생성과 관련이 있는 주요 유전자인 NOX4 및 catalase의 유전자발현 또한 유의적으로 감소하였다. 이 결과를 통해 연화 열수 및 에탄올 추출물이 3T3-L1 지방세포 내 중성지방의 축적 억제효과와 더불어 ROS 생성 억제에 효과적으로 작용함을 확인하였다. 따라서 연화 열수 및 에탄올 추출물은 비만과 같은 대사증후군 관련 질환의 개선을 위한 천연물 기능성 소재로의 활용이 기대된다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1996년도 제10회 식물생명공학심포지움 고등식물 발생생물학의 최근 진보
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• pp.26-38
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• 1996

To investigate the regulation of plant histone H2A gene expression, we isolated two H2A genes (TH254 and TH274) from wheat, which encode different types of variants. Both genes had an intron in the coding region. In the promoters, some characteristics sequences, such as Oct and Nona motifs, which are conserved among plant histone genes were also found, and they were located in a short region (about 120 bp) upstream from the putative TATA box. Analyses of promoter activity with H2A-GUS fusion genes in the transient system using tobacco protoplasts revealed novel types of positive cis-acting sequences in the TH254 promoter: a direct repeat of a 13-bp sequence (AGTTACATTATTG) and a stretch composed of an AT-rich sequence (ATATAGAAAATTAAAA) and a G-box (CACGTG). A quantitative S1 assay of the mRNA amounts from the TH254/GUS and TH274/GUG chimeric genes in stably transformed and cell cycle-synchronized tobacco cell lines showed that the promoters of both genes contained at least one cis-acting element responsible for S phase-specific expression. Histochemical analysis of transgenic tobacco plants carrying the chimeric genes showed that the promoters of the two H2A genes were both active in developing seedlings and flower organs but regulated in different manner.

• Journal of Plant Biotechnology
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• 제49권4호
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• pp.271-291
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• 2022

Pectin methylesterase (PME) plays an important role in vegetative and reproductive development and biotic/abiotic stress responses by regulating the degree of methyl-esterification of pectic polysaccharides in the plant cell wall. PMEs are encoded by a large multigene family in higher land plant genomes. In general, the expression of plant PME genes shows tissue- or cell-specific patterns and is induced by endogenous and exogenous stimuli. In this study, we identified PME multigene family members (CsPMEs) from the sweet orange genome and report detailed molecular characterization and expression profiling in different citrus tissues and two fruit developmental stages. We also discussed the possible functional roles of some CsPME genes by comparing them with the known functions of PMEs from other plant species. We identified 48 CsPME genes from the citrus genome. A phylogenetic tree analysis revealed that the identified CsPMEs were divided into two groups/types. Some CsPMEs showed very close phylogenetic relationships with the PMEs whose functions were formerly addressed in Arabidopsis, tomato, and maize. Expression profiling showed that some CsPME genes are highly or specifically expressed in the leaf, root, flower, or fruit. Based on the phylogenetic relationships and gene expression profiling results, we suggest that some CsPMEs could play functional roles in pollen development, pollen tube growth, cross incompatibility, root development, embryo/seed development, stomata movement, and biotic/abiotic stress responses. Our results shed light on the biological roles of individual CsPME isoforms and contribute to the search for genetic variations in citrus genetic resources.

• Journal of Plant Biotechnology
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• 제46권4호
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• pp.255-273
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• 2019

Orchids are indispensable to the floriculture industry due to their unique floral organization. The flowers have two outer whorls of tepals including a lip (labellum), and two inner whorls, pollinia and gynostemiun (column). The floral organization and development is controlled at the molecular level, mainly by the MADS-box gene family, comprising homeotic genes divided into type I and type II groups. The type I group has four sub-groups, Mα, Mβ, Mγ, and Mδ, playing roles in seed, embryo, and female reproductive organ development; the type II group genes form classes A, B, C, D, and E, which are a part of the MIKC^C subgroup with specific roles in florigenesis and organization. The coordinated functioning of these classes regulates the development of various floral whorls. The availability of genome and transcriptome sequence data for Phalaenopsis equestris offers an opportunity to validate the ABCDE model of flower development. Hence, this study sought to characterize the MADS-box gene family and elucidate of the ABCDE model. A total of 48 identified MADS-box proteins, including 20 type I [Mα (12), Mγ (8)] and 28 type II [MIKC^C (27), MIKC*(1)] members, were characterized for physico-chemical features and domains and motifs organization. The exon-intron distribution and the upstream cis-regulatory elements in the promoter regions of MADS-box genes were also analysed. The discrete pace of duplication events in type I and type II genes suggested differential evolutionary constraints between groups. The correlation of spatio-temporal expression pattern with the presence of specific cis-regulatory elements and putative protein-protein interaction within the different classes of MADS-box gene family endorse the ABCDE model of floral development.

• Plant Biotechnology Reports
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• 제3권3호
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• pp.183-190
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• 2009

Rho-related GTPase of plants (ROP) plays an important role in plant growth and development as a signaling protein. Plant RopGEFs are recently identified ROP activator proteins in Arabidopsis. In this study, we cloned 14 RopGEFs in Arabidopsis and characterized their expression patterns in response to abiotic stresses. Fourteen RopGEF genes were categorized into three groups based on their amino acid homologies and molecular sizes. Most RopGEFs were expressed predominantly in flower but some RopGEFs displayed a tissue-specific expression pattern. RopGEF1, 4, 5, and 11 were expressed in all tissues including root and leaves whereas RopGEF7, 8, 9, and 13 were expressed only in flowers. The transcript levels of 14 RopGEFs were changed significantly depending upon abiotic stresses such as cold, heat, drought and salts. RopGEF5 transcription was up-regulated by salt and drought treatment but down-regulated by heat. RopGEF14 transcript level was also increased by salt but decreased by heat stress. The transcript levels of RopGEF1, 7, 9, and 12 were enhanced in response to heat stress but showed no changes in response to cold stresses. Drought stress activated Group 3 RopGEFs such as RopGEF5 and 7. Taken together, 14 RopGEFs are responding to the abiotic stresses individually or as a group.