The asymmetric leaves2 ( as2) is a classical Arabidopsis thaliana (L.) Heynh. mutant that shows leaf lobes and leaflet-like structures from the petioles of leaves. Genetic and molecular analyses have demonstrated that...The asymmetric leaves2 ( as2) is a classical Arabidopsis thaliana (L.) Heynh. mutant that shows leaf lobes and leaflet-like structures from the petioles of leaves. Genetic and molecular analyses have demonstrated that the AS2 function is required for repression of meristem-related homeobox genes in leaves. In this study, we describe phenotypic characterizations of new as2 alleles that are in the Landsberg erecta (Ler) genetic background. In addition to the as2 phenotypes reported previously, the new as2 mutants have some leaves with petiole growth underneath the leaf blade, showing a lotus-leaf structure. More severe rosettes leaves of the as2 mutants form a filament-like structure, reflecting a loss of the adaxial-abaxial polarity in leaves. Among as2 mutants analyzed in different genetic backgrounds, only those that are in the Ler genetic background resulted in a high frequency of the lotus-leaf structure. We have isolated the AS2 gene by map-based gene cloning. The predicted AS2 protein contains a leucine-zipper motif, and its N-terminus shares high levels of sequence similarity to those of a group of predicted proteins with no known biological functions. AS2 transcripts were detected in leaves, flowers and fruits, but absent in stems, consistent with the mutant phenotypes.展开更多
Objective： The leaves of Chromolaena odorata, a highly invasive shrub found growing wild worldwide, are traditionally used for wound healing. Due to its high flavonoid contents, we aimed to find a new appli- cation f...Objective： The leaves of Chromolaena odorata, a highly invasive shrub found growing wild worldwide, are traditionally used for wound healing. Due to its high flavonoid contents, we aimed to find a new appli- cation for this plant. Preliminary tests using its ethanolic leaf extract showed that it could suppress the accumulation of lipids in adipocytes. We therefore studied the anti-adipogenic effect of several C. odorata leaf extracts and the relationship between molecular structure and bio-activity of its isolated flavonoid constituents using 3T3-LI preadipocytes/adipocytes as a model. Methods： Three leaf extracts and thirteen flavonoids isolated from C odorata were tested for their effect on lipid accumulation in 3T3-L1 adipocytes using AdipoRed reagent, with quercetin as the positive con- trol. The effects of active flavonoids on the adipocytes were confirmed by oil red O staining and visualized under a light microscope.Results： n-Hexane and ethyl acetate extracts of C odorata leaves displayed anti-adipogenic activity. The latter extract was the more potent one, especially at 40 μg/mL. Four flavonoids, pectolinarigenin, kaemp- feride, 4,2＇-dihydroxy-4＇,5＇,6＇-trimethoxychalcone and dillenetin, exhibited significant, concentration- dependent inhibitory effects on lipid accumulation in 3T3-L1 adipocytes. The most potent flavonoid obtained in this study was 4,2＇-dihydroxy-4＇,5＇,6＇-trimethoxychalcone, which caused 75% and 90% inhi- bition of cellular lipid accumulation at 30 and 50μmol/L, respectively. Both kaempferide and 4,2＇-dihy droxy-4＇,5＇,6＇-trimethoxychalcone were major constituents in the ethyl acetate extract of this plant.Conclusion： C odorata leaves contained several flavonoids with anti-adipogenic effects against lipid accu- mulation in 3T3-L1 adipocytes. The plant, normally considered a useless weed, may actually provide an abundant source of biologically active flavonoids.展开更多
Plants monitor daylength to synchronize their flowering time with their surroundings and thus maximize reproductive fitness.In Arabidopsis(Arabidopsis thaliana),CONSTANS(CO)activates the expression of FLOWERING LOCUS ...Plants monitor daylength to synchronize their flowering time with their surroundings and thus maximize reproductive fitness.In Arabidopsis(Arabidopsis thaliana),CONSTANS(CO)activates the expression of FLOWERING LOCUS T(FT);this activation is a crucial aspect of the daylength-dependent regulation of flowering time.Here,we demonstrate that the basic leucine zipper 3(bZIP3)transcription factor is important for CO-induced FT expression under long photoperiod conditions in Arabidopsis.We isolated bZIP3 as a CO-interacting protein by yeast two-hybrid screening and verified bZIP3-CO complex formation in Arabidopsis through coimmunoprecipitation assays.The temporal and spatial expression patterns of bZIP3 are very similar to those of CO,and bZIP3 protein levels fluctuate throughout the day,with high abundance in the late afternoon.The bzip3 mutant displayed delayed flowering under long photoperiods,whereas bZIP3 overexpression accelerated flowering regardless of daylength.bZIP3 directly binds to the FT promoter region containing CO-responsive elements in vivo.FT messenger RNA(m RNA)levels in the bzip3 mutant and bZIP3 overexpression lines correlated with their flowering times and changed only during the daytime.bZIP3 overexpression resulted in significantly lower FT transcript levels in the co mutant background than in the wild type.Furthermore,bZIP3 forms a complex with ASYMMETRIC LEAVES1(AS1),a CO partner that helps CO induce FT expression.The bzip3 as1 double mutant flowered later than the two single mutants under longer daylengths,and FT mRNA levels were much lower in the double mutant than in the bzip3 single mutant.Collectively,our findings uncover a new layer of photoperiod-dependent FT regulation in which bZIP3 facilitates CO to activate FT transcription by forming a complex with AS1.展开更多
During leaf organogenesis, a critical step for normal leaf primordium initiation is the repression of the class 1 KNOTTED1-like homeobox （KNOX） genes. After leaf primordia are formed, they must establish polarity fo...During leaf organogenesis, a critical step for normal leaf primordium initiation is the repression of the class 1 KNOTTED1-like homeobox （KNOX） genes. After leaf primordia are formed, they must establish polarity for normal leaf morphogenesis. Recent studies have led to the identification of a number of genes that participate in the class 1 KNOX gene repression and/or the leaf polarity establishment. ASTMMETRIC LEAVES1 and 2 （AS1 and AS2） are two of these genes, which are critical for both of these two processes. As a first step towards understanding the molecular genetic basis of the ASl-AS2 action, we determined the subcellular Iocalizations of the two proteins in both tobacco BY2 cells and Arabidopsis plants, by fusing them to yellow/cyan fluorescent protein （YFP/CFP）. Our data showed that AS1 and AS2 alone were predominantly localized in the nucleolus and the nucleoplasm, respectively. The presence of both AS1 and AS2 proteins in the same interphase cell demonstrated their co-localization in both nucleolus and nucleoplasm. In addition, AS1 alone was able to associate with the condensed chromosome in the metaphase cell. Our data suggest that AS1, AS2 and the ASl-AS2 protein complex may have distinct functions, which are all required for normal plant development.展开更多
Polycomb group proteins are important repressors of numerous genes in higher eukaryotes. However, the mechanism by which Polycomb group proteins are recruited to specific genes is poorly understood. In Arabidopsis, LI...Polycomb group proteins are important repressors of numerous genes in higher eukaryotes. However, the mechanism by which Polycomb group proteins are recruited to specific genes is poorly understood. In Arabidopsis, LIKE HETEROCHROMATIN PROTEIN 1 （LHP1）, also known as TERMINAL FLOWER 2, was originally proposed as a subunit of polycomb repressive complex 1 （PRC1） that could bind the tri-methylated lysine 27 of histone H3 （H3K27me3） established by the PRC2. In this work, we show that LHP1 mainly functions with PRC2 to establish H3K27me3, but not with PRC1 to catalyze monoubiquitination at lysine 119 of histone H2A. Our results show that complexes of the transcription factors ASYMMETRIC LEAVES 1 （AS1） and AS2 could help to establish the H3K27me3 modification at the chromatin regions of Class-I KNOTTED＇t-like homeobox （KNOX） genes BREVIPEDICELLU5 and KNAT2 via direct interactions with LHP1. Additionally, our transcriptome analysis indicated that there are probably more common target genes of AS1 and LHP1 besides Class-I KNOX genes during leaf development in Arabidopsis.展开更多
文摘The asymmetric leaves2 ( as2) is a classical Arabidopsis thaliana (L.) Heynh. mutant that shows leaf lobes and leaflet-like structures from the petioles of leaves. Genetic and molecular analyses have demonstrated that the AS2 function is required for repression of meristem-related homeobox genes in leaves. In this study, we describe phenotypic characterizations of new as2 alleles that are in the Landsberg erecta (Ler) genetic background. In addition to the as2 phenotypes reported previously, the new as2 mutants have some leaves with petiole growth underneath the leaf blade, showing a lotus-leaf structure. More severe rosettes leaves of the as2 mutants form a filament-like structure, reflecting a loss of the adaxial-abaxial polarity in leaves. Among as2 mutants analyzed in different genetic backgrounds, only those that are in the Ler genetic background resulted in a high frequency of the lotus-leaf structure. We have isolated the AS2 gene by map-based gene cloning. The predicted AS2 protein contains a leucine-zipper motif, and its N-terminus shares high levels of sequence similarity to those of a group of predicted proteins with no known biological functions. AS2 transcripts were detected in leaves, flowers and fruits, but absent in stems, consistent with the mutant phenotypes.
基金supported by a grant from Thailand Graduate Institute of Science and Technology(TGIST)(grant number:01-54-007)Nano-Cosmeceuticals Laboratory,National Nan-otechnology Center(NANOTEC)National Science and Technology Development Agency(NSTDA),Thailand
文摘Objective： The leaves of Chromolaena odorata, a highly invasive shrub found growing wild worldwide, are traditionally used for wound healing. Due to its high flavonoid contents, we aimed to find a new appli- cation for this plant. Preliminary tests using its ethanolic leaf extract showed that it could suppress the accumulation of lipids in adipocytes. We therefore studied the anti-adipogenic effect of several C. odorata leaf extracts and the relationship between molecular structure and bio-activity of its isolated flavonoid constituents using 3T3-LI preadipocytes/adipocytes as a model. Methods： Three leaf extracts and thirteen flavonoids isolated from C odorata were tested for their effect on lipid accumulation in 3T3-L1 adipocytes using AdipoRed reagent, with quercetin as the positive con- trol. The effects of active flavonoids on the adipocytes were confirmed by oil red O staining and visualized under a light microscope.Results： n-Hexane and ethyl acetate extracts of C odorata leaves displayed anti-adipogenic activity. The latter extract was the more potent one, especially at 40 μg/mL. Four flavonoids, pectolinarigenin, kaemp- feride, 4,2＇-dihydroxy-4＇,5＇,6＇-trimethoxychalcone and dillenetin, exhibited significant, concentration- dependent inhibitory effects on lipid accumulation in 3T3-L1 adipocytes. The most potent flavonoid obtained in this study was 4,2＇-dihydroxy-4＇,5＇,6＇-trimethoxychalcone, which caused 75% and 90% inhi- bition of cellular lipid accumulation at 30 and 50μmol/L, respectively. Both kaempferide and 4,2＇-dihy droxy-4＇,5＇,6＇-trimethoxychalcone were major constituents in the ethyl acetate extract of this plant.Conclusion： C odorata leaves contained several flavonoids with anti-adipogenic effects against lipid accu- mulation in 3T3-L1 adipocytes. The plant, normally considered a useless weed, may actually provide an abundant source of biologically active flavonoids.
基金supported partly by the grant from the National Institutes of Health Grant(No.R01GM079712 to T.I.)the Basic Science Research Program through the National Research Foundation(NRF)of Korea funded by the Ministry of Education(Nos.RS-2020-NR04950 and 2020R1A6A1A03044344 to W.S.C.)supported by the National Research Foundation(NRF)of Korea grant funded by the Korean government(MSIT)(No.RS-2024-00336624 to J.C.H.and No.2021R1A4A1032888 and No.RS-2025-00562751 to Y.H.S.)。
文摘Plants monitor daylength to synchronize their flowering time with their surroundings and thus maximize reproductive fitness.In Arabidopsis(Arabidopsis thaliana),CONSTANS(CO)activates the expression of FLOWERING LOCUS T(FT);this activation is a crucial aspect of the daylength-dependent regulation of flowering time.Here,we demonstrate that the basic leucine zipper 3(bZIP3)transcription factor is important for CO-induced FT expression under long photoperiod conditions in Arabidopsis.We isolated bZIP3 as a CO-interacting protein by yeast two-hybrid screening and verified bZIP3-CO complex formation in Arabidopsis through coimmunoprecipitation assays.The temporal and spatial expression patterns of bZIP3 are very similar to those of CO,and bZIP3 protein levels fluctuate throughout the day,with high abundance in the late afternoon.The bzip3 mutant displayed delayed flowering under long photoperiods,whereas bZIP3 overexpression accelerated flowering regardless of daylength.bZIP3 directly binds to the FT promoter region containing CO-responsive elements in vivo.FT messenger RNA(m RNA)levels in the bzip3 mutant and bZIP3 overexpression lines correlated with their flowering times and changed only during the daytime.bZIP3 overexpression resulted in significantly lower FT transcript levels in the co mutant background than in the wild type.Furthermore,bZIP3 forms a complex with ASYMMETRIC LEAVES1(AS1),a CO partner that helps CO induce FT expression.The bzip3 as1 double mutant flowered later than the two single mutants under longer daylengths,and FT mRNA levels were much lower in the double mutant than in the bzip3 single mutant.Collectively,our findings uncover a new layer of photoperiod-dependent FT regulation in which bZIP3 facilitates CO to activate FT transcription by forming a complex with AS1.
基金the Scientific and Technological Council Foundation of Shang-hai (04JC14017)the National Talent Training Fund in Basic Research of China (J0630643) to A. Dong+1 种基金the National Natural Science Foundation of China (30630041)the Chinese Academy of Sciences (KSCX2-YW-N-016) to H.Huang
文摘During leaf organogenesis, a critical step for normal leaf primordium initiation is the repression of the class 1 KNOTTED1-like homeobox （KNOX） genes. After leaf primordia are formed, they must establish polarity for normal leaf morphogenesis. Recent studies have led to the identification of a number of genes that participate in the class 1 KNOX gene repression and/or the leaf polarity establishment. ASTMMETRIC LEAVES1 and 2 （AS1 and AS2） are two of these genes, which are critical for both of these two processes. As a first step towards understanding the molecular genetic basis of the ASl-AS2 action, we determined the subcellular Iocalizations of the two proteins in both tobacco BY2 cells and Arabidopsis plants, by fusing them to yellow/cyan fluorescent protein （YFP/CFP）. Our data showed that AS1 and AS2 alone were predominantly localized in the nucleolus and the nucleoplasm, respectively. The presence of both AS1 and AS2 proteins in the same interphase cell demonstrated their co-localization in both nucleolus and nucleoplasm. In addition, AS1 alone was able to associate with the condensed chromosome in the metaphase cell. Our data suggest that AS1, AS2 and the ASl-AS2 protein complex may have distinct functions, which are all required for normal plant development.
基金supported by the National Basic Research Program of China (2012CB910500 and 2011CB944600)the National Natural Science Foundation of China (31370752)
文摘Polycomb group proteins are important repressors of numerous genes in higher eukaryotes. However, the mechanism by which Polycomb group proteins are recruited to specific genes is poorly understood. In Arabidopsis, LIKE HETEROCHROMATIN PROTEIN 1 （LHP1）, also known as TERMINAL FLOWER 2, was originally proposed as a subunit of polycomb repressive complex 1 （PRC1） that could bind the tri-methylated lysine 27 of histone H3 （H3K27me3） established by the PRC2. In this work, we show that LHP1 mainly functions with PRC2 to establish H3K27me3, but not with PRC1 to catalyze monoubiquitination at lysine 119 of histone H2A. Our results show that complexes of the transcription factors ASYMMETRIC LEAVES 1 （AS1） and AS2 could help to establish the H3K27me3 modification at the chromatin regions of Class-I KNOTTED＇t-like homeobox （KNOX） genes BREVIPEDICELLU5 and KNAT2 via direct interactions with LHP1. Additionally, our transcriptome analysis indicated that there are probably more common target genes of AS1 and LHP1 besides Class-I KNOX genes during leaf development in Arabidopsis.
