Lisha Shen

A Repressor Complex Governs the Integration of Flowering Signals in Arabidopsis

Multiple genetic pathways act in response to developmental cues and environmental signals to promote the floral transition, by regulating several floral pathway integrators. These include FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1). We show that the flowering repressor SHORT VEGETATIVE PHASE (SVP) is controlled by the autonomous, thermosensory, and gibberellin pathways, and directly represses SOC1 transcription in the shoot apex and leaf. Moreover, FT expression in the leaf is also modulated by SVP. SVP protein associates with the promoter regions of SOC1 and FT, where another potent repressor FLOWERING LOCUS C (FLC) binds. SVP consistently interacts with FLC in vivo during vegetative growth and their function is mutually dependent. Our findings suggest that SVP is another central regulator of the flowering regulatory network, and that the interaction between SVP and FLC mediated by various flowering genetic pathways governs the integration of flowering signals.

Regulation of Floral Patterning by Flowering Time Genes

Floral patterning in Arabidopsis requires activation of floral homeotic genes by the floral meristem identity gene, LEAFY (LFY). Here we show that precise activation of expression of class B and C homeotic genes in floral meristems is regulated by three flowering time genes, SHORT VEGETATIVE PHASE (SVP), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1), and AGAMOUS-LIKE 24 (AGL24), through direct control of a LFY coregulator, SEPALLATA3 (SEP3). Orchestrated repression of SEP3 by SVP, AGL24, and SOC1 is mediated by recruiting two interacting chromatin regulators, TERMINAL FLOWER 2/LIKE HETEROCHROMATIN PROTEIN 1 and SAP18, a member of SIN3 histone deacetylase complex. Our finding of coordinated regulation of SEP3 by flowering time genes reveals a hitherto unknown genetic pathway that prevents premature differentiation of floral meristems and determines the appropriate timing of floral organ patterning.

FTIP1 Is an Essential Regulator Required for Florigen Transport

FT-INTERACTING PROTEIN 1 is a novel protein that is involved in transporting florigen, a long-known mobile signal that induces flowering in plants in response to day length, from companion cells to sieve elements in the phloem of Arabidopsis.

Genome‐wide identification of SOC1 and SVP targets during the floral transition in Arabidopsis

Floral transition in Arabidopsis is tightly controlled by complex genetic regulatory networks in response to endogenous and environmental flowering signals. SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and SHORT VEGETATIVE PHASE (SVP), two key MADS-domain transcription factors, perceive these signals and function as antagonistic flowering regulators. To understand how these factors mediate floral transition, we mapped in vivo binding sites of SOC1 and SVP using chromatin immunoprecipitation followed by hybridization to whole-genome tiling arrays (ChIP-chip). Genes that encoded proteins with transcription regulator activity and transcription factor activity were the most enriched groups of genes of those bound by SOC1 and SVP, which indicates their central roles in flowering regulatory networks. In combination with gene expression microarray studies, we further identified the genes whose expression was controlled directly by SOC1 or SVP. Among the common direct targets identified, APETALA2 (AP2)-like genes that repress FT and SOC1 expression were down-regulated by SOC1, but up-regulated by SVP, revealing a complex feedback regulation among the key genes that determine the integration of flowering signals. SOC1 regulatory regions were also accessed by SOC1 itself and SVP, suggesting that self-activation and repression by SVP contribute to the control of SOC1 expression. In addition, ChIP-chip analysis demonstrated that miR156e and miR172a, which are involved in the regulation of AP2-like genes, were direct targets of SOC1 and SVP, respectively. Taken together, these findings revealed that feedback regulatory loops mediated by SOC1 and SVP are essential components of the gene regulatory networks that underpin the integration of flowering signals during floral transition.

Nuclear factor Y-mediated H3K27me3 demethylation of the SOC1 locus orchestrates flowering responses of Arabidopsis

Nuclear factor Y (NF-Y) is a conserved heterotrimeric transcription factor complex that binds to the CCAAT motifs within the promoter region of many genes. In plants, a large number of genes code for variants of each NF-YA, B or C subunit that can assemble in a combinatorial fashion. Here, we report the discovery of an Arabidopsis NF-Y complex that exerts epigenetic control over flowering time by integrating environmental and developmental signals. We show that NF-Y interacts with CONSTANS in the photoperiod pathway and DELLAs in the gibberellin pathway, to directly regulate the transcription of SOC1, a major floral pathway integrator. This NF-Y complex binds to a unique cis-element within the SOC1 promoter to modulate trimethylated H3K27 levels, partly through a H3K27 demethylase REF6. Our findings establish NF-Y complexes as critical mediators of epigenetic marks that regulate the response to environmental or intrinsic signals in plants.

Global Identification of DELLA Target Genes during Arabidopsis Flower Development

Gibberellin (GA) plays important roles in regulating many aspects of plant development. GA derepresses its signaling pathway by promoting the degradation of DELLA proteins, a family of nuclear growth repressors. Although the floral organ identity is established in flowers of the GA-deficient mutant ga1-3, the growth of all floral organs is severely retarded. In particular, abortive anther development in ga1-3 results in male sterility. Genetic analysis has revealed that various combinations of null mutants of DELLA proteins could gradually rescue floral organ defects in ga1-3 and that RGA is the most important DELLA protein involved in floral organ development. To elucidate the early molecular events controlled by RGA during flower development, we performed whole-genome microarray analysis to identify genes in response to the steroid-inducible activation of RGA in ga1-3 rgl2 rga 35S:RGA-GR. Although DELLA proteins were suggested as transcriptional repressors, similar numbers of genes were down-regulated or up-regulated by RGA during floral organ development. More than one-third of RGA down-regulated genes were specifically or predominantly expressed in stamens. A significant number of RGA-regulated genes are involved in phytohormone signaling or stress response. Further expression analysis through activation of RGA by steroid induction combined with cycloheximide identified eight genes as immediate targets of RGA. In situ hybridization and transgenic studies further showed that the expression pattern and function of several selected genes were consistent with the predictions from microarray analysis. These results suggest that DELLA regulation of floral organ development is modulated by multiple phytohormones and stress signaling pathways.

The J-Domain Protein J3 Mediates the Integration of Flowering Signals in Arabidopsis

This study demonstrates that Arabidopsis DNAJ HOMOLOG 3 (J3) responds to various flowering signals and mediates the transcriptional regulation of two major floral pathway integrators, FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1. J3 mediates this regulation through its interaction with a key flowering repressor, SHORT VEGETATIVE PHASE. The timing of the switch from vegetative to reproductive development in Arabidopsis thaliana is controlled by an intricate network of flowering pathways, which converge on the transcriptional regulation of two floral pathway integrators, FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1). SHORT VEGETATIVE PHASE (SVP) acts as a key flowering regulator that represses the expression of FT and SOC1. Here, we report the identification of another potent flowering promoter, Arabidopsis DNAJ HOMOLOG 3 (J3), which mediates the integration of flowering signals through its interaction with SVP. J3 encodes a type I J-domain protein and is ubiquitously expressed in various plant tissues. J3 expression is regulated by multiple flowering pathways. Loss of function of J3 results in a significant late-flowering phenotype, which is partly due to decreased expression of SOC1 and FT. We further show that J3 interacts directly with SVP in the nucleus and prevents in vivo SVP binding to SOC1 and FT regulatory sequences. Our results suggest a flowering mechanism by which J3 integrates flowering signals from several genetic pathways and acts as a transcriptional regulator to upregulate SOC1 and FT through directly attenuating SVP binding to their regulatory sequences during the floral transition.

The putative PRC1 RING-finger protein AtRING1A regulates flowering through repressing MADS AFFECTING FLOWERING genes in Arabidopsis

Polycomb group proteins play essential roles in the epigenetic control of gene expression in plants and animals. Although some components of Polycomb repressive complex 1 (PRC1)-like complexes have recently been reported in the model plant Arabidopsis, how they contribute to gene repression remains largely unknown. Here we show that a putative PRC1 RING-finger protein, AtRING1A, plays a hitherto unknown role in mediating the transition from vegetative to reproductive development in Arabidopsis. Loss of function of AtRING1A results in the late-flowering phenotype, which is attributed to derepression of two floral repressors, MADS AFFECTING FLOWERING 4/5 (MAF4/5), which in turn downregulate two floral pathway integrators, FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1. Levels of the H3K27me3 repressive mark at MAF4 and MAF5 loci, which is deposited by CURLY LEAF (CLF)-containing PRC2-like complexes and bound by LIKE HETEROCHROMATIN PROTEIN 1 (LHP1), are affected by AtRING1A, which interacts with both CLF and LHP1. Levels of the H3K4me3 activation mark correlate inversely with H3K27me3 levels at MAF4 and MAF5 loci. Our results suggest that AtRING1A suppresses the expression of MAF4 and MAF5 through affecting H3K27me3 levels at these loci to regulate the floral transition in Arabidopsis.

Emerging insights into florigen transport.

The photoperiodic control of flowering in plants begins with the perception of seasonal changes in day length and consequential induction of a mobile floral stimulus in leaves. This stimulus called florigen is transported from leaves to the shoot apical meristem to provoke the initiation of floral meristems. Decades of efforts have identified that the proteins encoded by FLOWERING LOCUS T (FT) in Arabidopsis and its orthologs in other plant species are part of the long-sought florigen. Emerging evidence suggests that long-distance transport of FT towards the shoot apical meristem occurs through the phloem in a regulated manner. This review summarizes the recent advances in understanding florigen transport and discusses the proven and potential regulators required for this process.

NaKR1 regulates long-distance movement of FLOWERING LOCUS T in Arabidopsis.

Flowering plants perceive photoperiodic signals in leaves to generate mobile stimuli required for the induction of flower formation at shoot apices. Although FLOWERING LOCUS T (FT) has been identified as part of the mobile floral stimuli in Arabidopsis thaliana, the mechanisms underlying long-distance movement of FT from leaves to shoot apices remain largely unclear. Here we show that a heavy-metal-associated (HMA) domain-containing protein, SODIUM POTASSIUM ROOT DEFECTIVE 1 (NaKR1), is activated by CONSTANS (CO) under long-day conditions and regulates long-distance movement of FT in Arabidopsis. Loss of function of NaKR1 compromises FT transport to shoot apices through sieve elements, causing late flowering under long-day conditions. NaKR1 and FT share similar expression patterns and subcellular localization, and interact with each other in vivo. Grafting experiments demonstrate that NaKR1 promotes flowering through mediating FT translocation from leaves to shoot apices. Thus, photoperiodic control of floral induction requires NaKR1-mediated long-distance delivery of florigenic signals.