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Low temperature-mediated repression and far-red light-mediated induction determine morning flowering locus t expression levels

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Journal of Integrative Plant Biology 2024年第1期66卷 103-120页

作者： Hayeon Kim Hye Won Kang Dae Yeon Hwang Nayoung Lee Akane Kubota takato Imaizumi Young Hun Song Department of Agricultural Biotechnology Seoul National UniversitySeoul 08826Korea Department of Biology Ajou UniversitySuwon 16499Korea Plant Genomics and Breeding Institute Seoul National UniversitySeoul 08826Korea Graduate School of Science and technology Nara Institute of Science and TechnologyIkomaNara 630‐0192Japan Department of Biology University of WashingtonSeattleWA 98195USA Institute of Agricultural Life Sciences Seoul National UniversitySeoul 08826Korea

In order to flower in the appropriate season,plants monitor light and temperature changes and alter downstream pathways that regulate florigen genes such as Arabidopsis(Arabidopsis thaliana)flowering locus t(Ft).In Arabidopsis,Ft messenger RNA levels peak in the morning and evening under natural long-day conditions(LDs).However,the regulatory mechanisms governing morning Ft induction remain poorly understood.the morning Ft peak is absent in typical laboratory LDs characterized by high red:far-red light(R:FR)ratios and constant temperatures.Here,we demonstrate that ZEItLUPE(ZtL)interacts with the Ft repressors tARGEt OF EAts(tOEs),thereby repressing morning Ft expression in natural environments.Under LDs with simulated sunlight(R:FR=1.0)and daily temperature cycles,which are natural LD-mimicking environmental conditions,Ft transcript levels in the ztl mutant were high specifically in the morning,a pattern that was mirrored in the toe1 toe2 double mutant.Low night-to-morning temperatures increased the inhibitory effect of ZtL on morning Ft expression by increasing ZtL protein levels early in the morning.Far-red light counteracted ZtL activity by decreasing its abundance(possibly via phytochrome A(phyA))while increasing GIGANtEA(GI)levels and negatively affecting the formation of the ZtL-GI complex in the morning.therefore,the phyA-mediated high-irradiance response and GI play pivotal roles in morning Ft induction.Our findings suggest that the delicate balance between low temperature-mediated ZtL activity and the far-red light-mediated functions of phyA and GI offers plants flexibility in fine-tuning their flowering time by controlling Ft expression in the morning.

关键词： far-red light flowering locus t flowering time GIGANtEA low night-to-morning temperature natural long days phytochrome A red to far-red ratio ZEItLUPE

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Molecular basis of CONStANS oligomerization in flowering locus t activation

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Journal of Integrative Plant Biology 2022年第3期64卷 731-740页

作者： Xiaolin Zeng Xinchen Lv Rui Liu Hang He Shiqi Liang Lixian Chen Fan Zhang Liu Chen Yuehui He Jiamu Du Peking-tsinghua Center for Life Sciences School of Advanced Agricultural SciencesPeking UniversityBeijing100871 China National Key Laboratory of Plant Molecular Genetics&Shanghai Center for Plant Stress Biology Chinese Academy of Sciences Center for Excellence in Molecular Plant SciencesShanghai201602 China Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes Institute of Plant and Food ScienceDepartment of BiologySchool of Life SciencesSouthern University of Science and TechnologyShenzhen518055 China Peking University Institute of Advanced Agricultural Sciences Weifang261000 China

the transcription factor CONStANS(CO)integrates day-length information to induce the expression of florigen flowering locus t(Ft)in Arabidopsis.We recently reported that the C-terminal CCt domain of CO forms a complex with NUCLEAR FACtOR-YB/YC to recognize multiple cis-elements in the Ft promoter,and the N-terminal tandem B-box domains form a homomultimeric assembly.However,the mechanism and biological function of CO multimerization remained unclear.Here,we report that CO takes on a head-to-tail oligomeric configuration via its B-boxes to mediate Ft activation in long days.the crystal structure of B-boxesCOreveals a closely connected tandem B-box fold forming a continuous head-to-tail assembly through unique CDHH zinc fingers.Mutating the key residues involved in CO oligomerization resulted in a non-functional CO,as evidenced by the inability to rescue co mutants.By contrast,a transgene encoding a human p53-derived tetrameric peptide in place of the B-boxesCOrescued co mutant,emphasizing the essential role of BboxesCO-mediated oligomerization.Furthermore,we found that the four tGtG-bearing cis-elements in Ft proximal promoter are required for Ft activation in long days.Our results suggest that CO forms a multimer to bind to the four tGtG motifs in the Ft promoter to mediate Ft activation.

关键词： B-box domain CONStANS crystal structure flowering locus t photoperiod

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Photoperiodic flowering in Arabidopsis:Multilayered regulatory mechanisms of CONStANS and the florigen flowering locus t

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Plant Communications 2023年第3期4卷 47-65页

作者： Hiroshi takagi Andrew K.Hempton takato Imaizumi Department of Biology University of WashingtonSeattleWA 98195-1800USA Center for Gene Research Nagoya UniversityNagoya 464-8602Japan

the timing of flowering affects the success of sexual reproduction.this developmental event also determines crop yield,biomass,and longevity.therefore,this mechanism has been targeted for improvement along with crop domestication.the underlying mechanisms of flowering are highly conserved in angiosperms.Central to these mechanisms is how environmental and endogenous conditions control transcriptional regulation of the flowering locus t(Ft)gene,which initiates floral development under long-day conditions in Arabidopsis.Since the identification of Ft as florigen,efforts have been made to understand the regulatory mechanisms of Ft expression.Although many transcriptional regulators have been shown to directly influence Ft,the question of how they coordinately control the spatiotemporal expression patterns of Ft still requires further investigation.Among Ft regulators,CONStANS(CO)is the primary one whose protein stability is tightly controlled by phosphorylation and ubiquitination/proteasome-mediated mechanisms.In addition,various CO interaction partners,some of them previously identified as Ft transcriptional regulators,positively or negatively modulate CO protein activity.the Ft promoter possesses several transcriptional regulatory"blocks,"highly conserved regions among Brassicaceae plants.Different transcription factors bind to specific blocks and affect Ft expression,often causing topological changes in Ft chromatin structure,such as the formation of DNA loops.We discuss the current understanding of the regulation of Ft expression mainly in Arabidopsis and propose future directions related to this topic.

关键词： photoperiodic flowering Arabidopsis thaliana circadian clock photoreceptor CONStANS flowering locus t

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ERF1 delays flowering through direct inhibition of flowering locus t expression in Arabidopsis

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Journal of Integrative Plant Biology 2021年第10期63卷 1712-1723页

作者： Yanli Chen Liping Zhang Haiyan Zhang Ligang Chen Diqiu Yu CAS Key Laboratory of tropical Plant Resources and Sustainable Use Xishuangbanna Tropical Botanical GardenChinese Academy of SciencesMengla 666303China University of Chinese Academy of Sciences Beijing 100049China Key Laboratory for Plant Diversity and Biogeography of East Asia Kunming Institute of BotanyChinese Academy of SciencesKunming 650201China Center of Economic Botany Core Botanical GardensChinese Academy of SciencesMenglunMengla 666303China State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan Yunnan UniversityKunming 650091China

EtHYLENE RESPONSE FACtOR1(ERF1)is a key component in ethylene signaling,playing crucial roles in both biotic and abiotic stress responses.Here,we demonstrate that ERF1 also has an important role during floral initiation in Arabidopsis thaliana.Knockdown or knockout of ERF1 accelerated floral initiation,whereas overexpression of ERF1 dramatically delayed floral transition.these contrasting phenotypes were correlated with opposite transcript levels of flowering locus t(Ft).Chromatin immunoprecipitation(ChIP)assays revealed that ERF1 associates with genomic regions of the Ft gene to repress its transcription.ft-10/ERF1RNAi plants showed a similar flowering phenotype to the ft-10 mutant,whereas the flowering of Ftox/ERF1ox mimicked that of Ftox plants,suggesting that ERF1 acts upstream of Ft during floral initiation.Similarly,altered floral transition in ethylene-related mutants was also correlated with Ft expression.Further analysis suggested that ERF1 also participates in delay in flowering-time control mediated by the ethylene precursor 1-aminocyclopropane-1-carboxylic acid.thus,ERF1 may act as a negative modulator of flowering-time control by repressing Ft transcription in Arabidopsis.

关键词： Arabidopsis thaliana ethylene EtHYLENE RESPONSE FACtOR1 floral initiation flowering locus t

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Long-distance control of potato storage organ formation by SELF PRUNING 3D and flowering locus t-like 1

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Plant Communications 2023年第3期4卷 258-273页

作者： SaloméPratand Botao Song Shenglin Jing Peng Jiang Xiaomeng Sun Liu Yu Enshuang Wang Jun Qin Fei Zhang SaloméPrat Botao Song National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops Huazhong Agricultural UniversityWuhanHubei 430070China Key Laboratory of Potato Biology and Biotechnology(HZAU) Ministry of Agriculture and Rural AffairsWuhanHubei 430070China College of Horticulture and Forestry Science Huazhong Agricultural UniversityWuhanHubei 430070China Centro de Investigación en Agrigenomica(CRAG) CSIC-IRTA-UAB-UBCerdanyola08193 BarcelonaSpain

Plants program their meristem-associated developmental switches for timely adaptation to a changing environment.Potato(Solanum tuberosum L.)tubers differentiate from specialized belowground branches or stolons through radial expansion of their terminal ends.During this process,the stolon apex and closest axillary buds enter a dormancy state that leads to tuber eyes,which are reactivated the following spring and generate a clonally identical plant.the potato flowering locus t homolog SELF-PRUNING 6A(StSP6A)was previously identified as the major tuber-inducing signal that integrates day-length cues to control the storage switch.However,whether some other long-range signals also act as tuber organogenesis stimuli remains unknown.Here,we show that the florigen SELF PRUNING 3D(StSP3D)and flowering locus t-like 1(StFtL1)genes are activated by short days,analogously to StSP6A.Overexpression of StSP3D or StFtL1 promotes tuber formation under non-inductive long days,and the tuber-inducing activity of these proteins is graft transmissible.Using the non-tuber-bearing wild species Solanum etuberosum,a natural SP6A null mutant,we show that leaf-expressed SP6A is dispensable for StSP3D long-range activity.StSP3D and StFtL1 mediate secondary activation of StSP6A in stolon tips,leading to amplification of this tuberigen signal.StSP3D and StFtL1 were observed to bind the same protein partners as StSP6A,suggesting that they can also form transcriptionally active complexes.together,our findings show that additional mobile tuber-inducing signals are regulated by the photoperiodic pathway.

关键词： potato Solanum tuberosum L. flowering locus t StSP3D tuberization tuber-inducing signal long-distance signaling

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烟草flowering locus t基因鉴定注释及分析

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分子植物育种 2022年

作者：杨帅张立猛牛永志潘磊郑昀晔玉溪中烟种子有限责任公司云南省烟草农业科学研究院

flowering locus t (Ft)基因是影响高等植物开花的关键基因，其翻译产生的蛋白产物是一种可长距离转运的成花激素，对开花时间起着重要的作用。目前在烟草种子生产中，均采用杂交育种方式，因此掌握烟草的开花时间尤为重要。为更好地了... 详细信息

flowering locus t (Ft)基因是影响高等植物开花的关键基因，其翻译产生的蛋白产物是一种可长距离转运的成花激素，对开花时间起着重要的作用。目前在烟草种子生产中，均采用杂交育种方式，因此掌握烟草的开花时间尤为重要。为更好地了解Ft开花基因在烟草中的结构、表达及其进化关系，本研究利用8个烟草种的完整基因组序列数据，鉴定并分析了Ft基因在8个烟草种中的蛋白结构、保守结构域、进化关系、共线性、基因注释和基因表达。结果显示，在8个烟草基因组中筛选鉴定出106个Ft基因，并将这些Ft基因分为6个组。在与其他植物Ft基因的进化分析中发现野生烟草Ft基因在进化过程中与茄科植物较接近。对普通烟草和两个亲本野生烟草的共线性分析，表明了Nicotiana tabacum BX和Nicotiana tabacum tN90的Ft基因进化关系非常接近。在Ft基因注释分析中，发现有22个基因是促进烟草花朵发育，有16个基因是参与抑制花发育的生物学途径。所有106个Ft基因均参与Ko04712植物昼夜节律代谢途径，并在其中担任开花途径的一部分。Ft基因在Nicotiana tabacum tN90的9个不同生长发育时期的表达模式分析发现，Ft基因在根、茎、叶和花中均有表达，单表达量存在明显差异，其中大部分的Ft基因是表达于鲜花朵、成熟花朵和衰老花朵3个时期中。这些结果将对以后研究中的基因查询、基因克隆和功能验证等方面发挥重要的作用。

关键词： flowering locus t 注释共线性基因表达生物信息学

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the Chromosome-Level Genome Sequence of the Autotetraploid Alfalfa and Resequencing of Core Germplasms Provide Genomic Resources for Alfalfa Research

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Molecular Plant 2020年第9期13卷 1250-1261页

作者： Chen Shen Huilong Du Zhuo Chen Hongwei Lu Fugui Zhu Hong Chen Xiangzhao Meng Qianwen Liu Peng Liu Lihua Zheng Xiuxiu Li Jiangli Dong Chengzhi Liang tao Wang State Key Laboratory of Agrobiotechnology College of Biological SciencesChina Agricultural UniversityBeijingChina State Key Laboratory of Plant Genomics Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijingChina University of Chinese Academy of Sciences BeijingChina

Alfalfa(Medicago sativa)is one of the most important forage crops in the world;however,its molecular genetics and breeding research are hindered due to the lack of a high-quality reference genome.Here,we report a de novo assembled 816-Mb high-quality,chromosome-level haploid genome sequence for‘Zhongmu No.1’alfalfa,a heterozygous autotetraploid.the contig N50 is 3.92 Mb,and 49165 genes are annotated in the genome.the alfalfa genome is estimated to have diverged from M.truncatula approximately 8 million years ago.Genomic population analysis of 162 alfalfa accessions revealed high genetic diversity,weak population structure,and extensive gene flow from wild to cultivated alfalfa.Genome-wide association studies identified many candidate genes associated with important agronomic traits.Furthermore,we showed that MsFta2,a flowering locus t homolog,whose expression is upregulated in salt-resistant germplasms,may be associated with fall dormancy and salt resistance.taken together,these genomic resources will facilitate alfalfa genetic research and agronomic improvement.

关键词： alfalfa genome assembly population gen etics GW AS flowering locus t

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Genetic and Epigenetic Understanding of the Seasonal timing of flowering

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Plant Communications 2020年第1期1卷 25-37页

作者： Yuehui He tao Chen Xiaolin Zeng National Key Laboratory of Plant Molecular Genetics&Shanghai Center for Plant Stress Biology Chinese Academy of Sciences Center for Excellence in Molecular Plant SciencesShanghai 201602China

the developmental transition to flowering in many plants is timed by changing seasons,which enables plants to flower at a season that is favorable for seed production.Many plants grown at high latitudes perceive the seasonal cues of changing day length and/or winter cold(prolonged cold exposure),to regulate the expression of flowering-regulatory genes through the photoperiod pathway and/or vernalization pathway,and thus align flowering with a particular season.Recent studies in the model flowering plant Arabidopsis thaliana have revealed that diverse transcription factors engage various chromatin modifiers to regulate several key flowering-regulatory genes including flowering locus C(FLC)and flowering locus t(Ft)in response to seasonal signals.Here,we summarize the current understanding of molecular and chromatin-regulatory or epigenetic mechanisms underlying the vernalization response and photoperiodic control of flowering in Arabidopsis.Moreover,the conservation and divergence of regulatory mechanisms for seasonal flowering in crops and other plants are briefly discussed.

关键词： vernalization photoperiodism flowering time flowering locus C flowering locus t chromatin modification

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四季金花茶和金花茶成花诱导差异的分子机制探究

四季金花茶和金花茶成花诱导差异的分子机制探究

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作者：崔佳广西大学

学位级别：硕士

金花茶组(Sect.Chrysanthae)植物具有很高的观赏价值,大多数金花茶组植物每年单次开花,而四季金花茶(Camellia perpetua)每年多次开花。为探究四季金花茶(C.perpetua)和金花茶(C.petelotii)成花机制的差异,本文首先对两种金花茶不同季... 详细信息

金花茶组(Sect.Chrysanthae)植物具有很高的观赏价值,大多数金花茶组植物每年单次开花,而四季金花茶(Camellia perpetua)每年多次开花。为探究四季金花茶(C.perpetua)和金花茶(C.petelotii)成花机制的差异,本文首先对两种金花茶不同季节的花芽分化及开花特性进行了比较;通过转录组学分析四季金花茶花芽分化高峰期和低谷期的顶芽及花芽的差异表达基因,并对Cper CEN1基因进行功能研究;通过GA等植物激素处理两种金花茶的顶芽,比较不同激素对两种金花茶顶芽的休眠及生长发育的影响;克隆两种金花茶Ft基因和启动子并转化拟南芥,分析比较两种金花茶Ft的功能,探讨Ft与两种金花茶成花习性差异的关系。获得以下研究结果:(1)统计两种金花茶一年至多年生枝的花芽着生情况,发现两种金花茶大部分的花芽着生于一年生枝条上,四季金花茶花芽主要着生于顶芽基部,由顶芽基部鳞片的腋芽分化形成,金花茶当年生枝条均有花芽形成,顶芽基部鳞片虽有腋芽,但不可以分化成花芽。对两种金花茶顶芽和花芽进行解剖、石蜡切片和扫描电镜观察,结果显示两种金花茶不同时期的顶芽的顶端分生组织以及分化起始阶段的花芽的花原基、苞片原基形态均无明显差异,暗示两种金花茶成花习性的差异可能是有成花诱导引起的。(2)对四季金花茶花芽分化高峰期(3月)和低谷期(9月)的顶芽与花芽进行比较转录组学研究,筛选得到11 067个显著差异表达的基因,其中628个基因在花芽中特异性表达,另有151个基因在花芽中特异性上调表达。通过KEGG和GO富集分析,差异基因主要集中在淀粉蔗糖代谢、植物激素合成代谢以及信号传导等通路上,共鉴定了37个成花诱导相关的功能基因。对PEBP家族基因的表达模式分析发现,四季金花茶Cper CEN1在3月和9月的顶芽中差异显著表达,而金花茶Cpet CEN1在顶芽中几乎没有表达。进一步研究发现,两种金花茶CEN1基因组序列完整,未存在功能缺失的情况,过表达四季金花茶Cper CEN1导致拟南芥推迟开花,并影响花序发育,说明两种金花茶成花机制的差异不是由于CEN1引起的,四季金花茶存在独特的成花诱导机制。(3)通过GA、PAC、生长素、6-BA、玉米素、反式玉米素和乙烯利等激素处理两种金花茶的顶芽,比较不同激素对两种金花茶顶芽的休眠、生长以及花芽分化的影响。结果发现,四季金花茶盛花期后,GA(200 mg/L)处理均促进两种金花茶的顶芽提前抽稍,而冬季不同激素处理的顶芽均未出现抽稍等现象。在冬季将两种金花茶移入温室中生长(平均温度比自然环境高约10°C),诱导四季金花茶提前进行花芽分化,同样条件下用PAC处理会抑制成花,测定结果发现可溶性糖含量与成花关系密切。(4)四季金花茶Cper Ft和金花茶Cpet Ft基因序列中均包含4个外显子与3个内含子;Ft同源基因核苷酸序列相似度高达91%,氨基酸序列仅存在1位的氨基酸差异且不是功能位点。克隆两种金花茶Ft基因上游2.5 kb序列,相似度为98%。为比较两种金花茶Ft表达特性和功能特性,构建Cper Ftpro::GUS和Cpet Ftpro::GUS、35S::Cper Ft、g Cper Ft和g Cpet Ft 5个表达载体,分别转化拟南芥Col-0和ft-10。结果显示,35S::Cper Ft促进拟南芥极早开花,且可以回补ft-10;Cper Ftpro::GUS和Cpet Ftpro::GUS转化拟南芥Col-0,在叶片、花序、花朵和角果中均有表达,但无明显的表达模式差异。g Cper Ft和g Cpet Ft回补ft-10没回补成功,但产生抽双薹的新表型。四季金花茶成花素基因Ft是否和金花茶存在差异,是否直接参与其多次开花,仍需进一步研究。

关键词：四季金花茶金花茶 CENtRORADIALIS 植物激素 flowering locus t 成花诱导

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成花素基因PdFt的克隆及其对牡丹成花的影响

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中国农业科学 2014年第13期47卷 2613-2624页

作者：朱富勇刘传娇薛璟祺王顺利张萍任秀霞张秀新中国农业科学院蔬菜花卉研究所/农业部园艺作物生物学与种质创制重点实验室

【目的】克隆紫牡丹Ft同源基因,分析其表达模式以及对紫牡丹成花的调控作用。【方法】以紫牡丹为试验材料,通过Rt-PCR的方法克隆紫牡丹Ft同源基因PdFt。通过实时荧光定量PCR分析PdFt在紫牡丹不同组织、紫牡丹开花物候期各过程及不同处... 详细信息

【目的】克隆紫牡丹Ft同源基因,分析其表达模式以及对紫牡丹成花的调控作用。【方法】以紫牡丹为试验材料,通过Rt-PCR的方法克隆紫牡丹Ft同源基因PdFt。通过实时荧光定量PCR分析PdFt在紫牡丹不同组织、紫牡丹开花物候期各过程及不同处理的表达情况,并探讨PdFt与花芽发育状态的关系。同时,将克隆到的紫牡丹PdFt克隆到表达载体pEt-28a上,构建融合表达载体pEt-28a-PdFt,转化到大肠杆菌BL21(DE3)并诱导表达。【结果】克隆得到包括完整开放阅读框(ORF)的PdFt cDNA序列,ORF长度为522 bp,编码173个氨基酸,GenBank登录号为KF113360。氨基酸序列比对表明:PdFt蛋白具有一个典型的PEBP结构域,属于PEBP家族;并且与GenBank中已克隆的Ft同源基因具有高度的同源性。实时荧光定量PCR分析结果表明,PdFt在紫牡丹根、茎、叶、芽中均有表达,芽中表达量最高,叶中表达量最低。紫牡丹开花物候期各过程PdFt的表达分析表明,在花芽膨大期表达量最高,随着花蕾的发育,PdFt的表达量逐渐降低。PdFt在不同光照温度条件下的表达分析表明,短日照和低温处理均抑制PdFt基因的表达。不同状态花蕾中PdFt的表达分析结果表明,败育花蕾PdFt的表达量低于正常花蕾。同时,GA3及去叶处理均能提高PdFt的表达量。所构建的原核表达载体,经IPtG诱导和SDS-PAGE电泳检测结果表明,表达蛋白与预期蛋白大小一致。【结论】紫牡丹PdFt与已克隆的Ft同源基因高度同源,属于Ft亚家族,在紫牡丹顶芽中表达量最高,可能调控牡丹开花,PdFt的克隆及表达分析为研究紫牡丹成花的分子机理奠定了基础。

关键词：紫牡丹 flowering locus t 实时定量PCR 原核表达

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