Analysis of flowering pathway integrators in Arabidopsis

Flowering is regulated by an integrated network of several genetic pathways in Arabidopsis. The key genes integrating multiple flowering pathways are FT, SOC1 and LFY. To elucidate the interactions among these integrators, genetic analyses were performed. FT and SOC1 share the common upstream regulators CO, a key component in the long day pathway, and FLC, a flowering repressor integrating autonomous and vernalization pathways. However, the soc1 mutation further delayed the flowering time of long day pathway mutants including ft, demonstrating that SOC1 acts partially independently of FT. Although soc1 did not show an obvious defect in flower meristem determination on its own, it dramatically increased the number of coflorescences in a lfy mutant, which is indicative of a defect in floral initiation. Therefore, double mutant analysis shows that the three integrators have both overlapping and independent functions in the determination of flowering time and floral initiation. The expression analysis showed that FT regulates SOC1 expression, and SOC1 regulates LFY expression, but not vice versa, which is consistent with the fact that FT and LFY have the least overlapping functions among the three integrators. The triple mutation ft soc1 lfy did not block flowering completely under long days, indicating the presence of other integrators. Finally, vernalization accelerated flowering of flc ft soc1 and ft soc1 lfy triple mutants, which shows that the vernalization pathway also has targets other than FLC, FT, SOC1 and LFY. Our genetic analysis reveals the intricate nature of genetic networks for flowering.     

MicroRNA-target Interactions: Important Signaling Modules Regulating Flowering Time in Diverse Plant Species

Successful reproduction of flowering plants requires the appropriate timing of the floral transition, as triggered by environmental and internal cues and as regulated by multiple signaling modules. Among these modules, microRNAs (miRNAs), the evolutionarily conserved regulators, respond to environmental and internal cues and network with other integrators of flowering cues. Moreover, miRNA signaling modules affect the timing of flowering in many plant species. Here, we comprehensively review recent progress in understanding the function of miRNAs and their target genes in flowering time regulation in diverse plant species. We focus on the role of the miRNA-target gene modules in various flowering pathways and their conserved and divergent functions in flowering plants. We also examine, in depth, the crosstalk by sequential activity of miR156 and miR172, two of the most-studied and evolutionarily conserved miRNAs in both annual and perennial plants.     

The rhythmic expression of genes controlling flowering time in southern and northern populations of invasive Ambrosia artemisiifolia

Aims Flowering time has been suggested to be an important adaptive trait during the dispersal of invasive species, and identifying the molecular mechanisms underlying flowering time may provide insight into the local adaptation during the process of invasion. Here, we conducted a preliminary exploration on the genetic basis of the differentiation of flowering time in Ambrosia artemisiifolia .Methods Using relative real-time fluorescent quantitative polymerase chain reaction, we investigated the expression levels of eight flowering-related genes, including AP1, FT, SOC1, CRY2, FKF1, GI, CO2 and SPY, in leaves and flowers at different time points in individuals from northern Beijing and southern Wuhan populations that exhibit significant differences in flowering times to identify any rhythmic changes in gene expression and their association with differential flowering times.Important findings The differentiation of flowering time in the A. artemisiifolia populations was closely associated with five genes involved in flowering pathways. The floral pathway integrators FT and SOC1 and floral meristem identity gene AP1 exhibited increased expression during flowering. The photoreceptor CRY2 in the light-dependent pathway and the SPY gene in the gibberellin pathway displayed specific expression patterns over time. In earlier-flowering Beijing plants, CRY2 expression was lower and SPY expression was higher than in Wuhan plants. The expression patterns of these five genes suggest a molecular basis for the differentiation of flowering time in A. artemisiifolia .     

Flowering time regulation produces much fruit

Many of the molecular details regarding the promotion of flowering in response to prolonged exposure to cold temperatures (vernalization) and daylength have recently been elucidated in Arabidopsis. The daylength and vernalization pathway converge in the regulation of floral promoters referred to as floral integrators. In the meristem, vernalization promotes flowering through the epigenetic repression of the floral repressor FLOWERING LOCUS C. This allows for the induction of floral integrators by CONSTANS under inductive long days. In the vasculature of leaves, CONSTANS protein is produced only in long days where it acts to promote the expression of FLOWERING LOCUS T (FT). FT protein is then translocated to the meristem where it acts to promote floral induction. Thus a detailed molecular framework for the regulation of flowering time has now been established in Arabidopsis.     

Integration of floral inductive signals by flowering locus T and suppressor of overexpression of Constans 1

Genetic analyses have identified four major genetic pathways which control the timing of floral transitions in Arabidopsis . The floral promotion or inhibitory signals from these distinct pathways ultimately converge into a subset of genes, commonly known as the floral integrators, such that the plants are compelled either to switch into a flowering phase or to remain in the vegetative stage. In this review, we have discussed the recent findings regarding cross-talk between floral promotion pathways and flowering locus T and suppressor of overexpression of Constans 1 , the major floral integrators within Arabidopsis . The spatial regulation of these integrators has also been discussed, as these spatial patterns may help us to understand the manner in which floral inductive signals are transmitted. We have also discussed conservation and divergence with regard to the regulation of floral integrators between Arabidopsis and rice. Finally, we have provided a series of insights into the complex signaling network which coordinates plant development.     

Heat shock protein 101 (HSP101) promotes flowering under nonstress conditions

Heat shock proteins (HSPs) are stress-responsive proteins that are conserved across all organisms. Heat shock protein 101 (HSP101) has an important role in thermotolerance owing to its chaperone activity. However, if and how it functions in development under nonstress conditions is not yet known. By using physiological, molecular, and genetic methods, we investigated the role of HSP101 in the control of flowering in Arabidopsis (Arabidopsis thaliana (L.) Heynh.) under nonstress conditions. Knockout and overexpression of HSP101 cause late and early flowering, respectively. Late flowering can be restored by rescue of HSP101. HSP101 regulates the expression of genes involved in the six known flowering pathways; the most negatively regulated genes are FLOWERING LOCUS C (FLC) and SHORT VEGETATIVE PHASE (SVP); downstream integrators of the flowering pathways are positively regulated. The late-flowering phenotype of loss-of-HSP101 mutants is suppressed by both the mutations of FLC and SVP. The responses of flowering time to exogenous signals do not change in HSP101 mutants. HSP101 is also found in nonspecific regions according to subcellular localization. We found that HSP101 promotes flowering under nonstress conditions and that this promotion depends on FLC and SVP. Our data suggest that this promotion could occur through a multiple gene regulation mechanism.

Heat shock protein 101 promotes flowering under nonstress conditions in Arabidopsis (Arabidopsis thaliana (L.) Heynh.).     

光周期途径核心因子CO的开花调控机制

CONSTANS（CO）基因是生物钟和开花时间基因之间监测日照长度的重要元件,在光周期途径中发挥核心功能。CO可以整合光信号和生物钟信号,诱导开花途径整合子FLOWERINGLOCUST（F即和SUPPRESSOROF OVEREXPRESSION OF CONSTANS 1（SOC1）的表达,进而促进植株开花。本文综述CO基因的开花调控机制,并结合CO基因的研究现状展望了其未来的研究方向。     

UGT87A2, an Arabidopsis glycosyltransferase, regulates flowering time via FLOWERING LOCUS C

? Family 1 glycosyltransferases comprise the greatest number of glycosyltransferases found in plants. The widespread occurrence and diversity of glycosides throughout the plant kingdom underscore the importance of these glycosyltransferases. ? Here, we describe the identification and characterization of a late-flowering Arabidopsis (Arabidopsis thaliana) mutant, in which a putative family 1 glycosyltransferase gene, UGT87A2, was disrupted. The role and possible mechanism of UGT87A2 in the regulation of flowering were analyzed by molecular, genetic and cellular approaches. ? The ugt87a2 mutant exhibited late flowering in both long and short days, and its flowering was promoted by vernalization and gibberellin. Furthermore, the mutant flowering phenotype was rescued by the wild-type UGT87A2 gene in complementation lines. Interestingly, the expression of the flowering repressor FLOWERING LOCUS C was increased substantially in the mutant, but decreased to the wild-type level in complementation lines, with corresponding changes in the expression levels of the floral integrators and floral meristem identity genes. The expression of UGT87A2 was developmentally regulated and its protein products were distributed in both cytoplasm and nucleus. ? Our findings imply that UGT87A2 regulates flowering time via the flowering repressor FLOWERING LOCUS C. These data highlight an important role for the family 1 glycosyltransferases in the regulation of plant flower development.     

Dissection of floral induction pathways using global expression analysis

Flowering of the reference plant Arabidopsis thaliana is controlled by several signaling pathways, which converge on a small set of genes that function as pathway integrators. We have analyzed the genomic response to one type of floral inductive signal, photoperiod, to dissect the function of several genes transducing this stimulus, including CONSTANS, thought to be the major output of the photoperiod pathway. Comparing the effects of CONSTANS with those of FLOWERING LOCUS T, which integrates inputs from CONSTANS and other floral inductive pathways, we find that expression profiles of shoot apices from plants with mutations in either gene are very similar. In contrast, a mutation in LEAFY, which also acts downstream of CONSTANS, has much more limited effects. Another pathway integrator, SUPPRESSOR OF OVEREXPRESSION OF CO 1, is responsive to acute induction by photoperiod even in the presence of the floral repressor encoded by FLOWERING LOCUS C. We have discovered a large group of potential floral repressors that are down-regulated upon photoperiodic induction. These include two AP2 domain-encoding genes that can repress flowering. The two paralogous genes, SCHLAFMUTZE and SCHNARCHZAPFEN, share a signature with partial complementarity to the miR172 microRNA, whose precursor we show to be induced upon flowering. These and related findings on SPL genes suggest that microRNAs play an important role in the regulation of flowering.     

PbCOL8 is a clock-regulated flowering time repressor in pear

The floral transition is controlled by diverse endogenous and exogenous cues. In many species, COL (CONSTANS-like) genes integrate light and circadian clock signals to regulate flowering time. However, little is known about COLs in perennial woody plants. Here, we identified 15 PbCOLs in pear (Pyrus bretschneideri). PbCOLs were classified into three groups by phylogenetic tree analysis using protein sequences. Multiple sequence alignment analysis revealed conserved B-box and CCT (CO, CO-like, and TOC1) domains in all PbCOL members. This result suggested that PbCOLs might possess conserved functions as other species. Six PbCOLs were found to be regulated by both circadian clock and photoperiod. Here, we showed that PbCOL8, a member of group 2, suppressed the flowering signal integrators FT and SOC1 and could repress flowering time. These findings will contribute to elucidation of the mechanism of floral initiation in pear.     

Understanding the genetic and epigenetic architecture in complex network of rice flowering pathways

Although the molecular basis of flowering time control is well dissected in the long day (LD) plant Arabidopsis, it is still largely unknown in the short day (SD) plant rice. Rice flowering time (heading date) is an important agronomic trait for season adaption and grain yield, which is affected by both genetic and environmental factors. During the last decade, as the nature of florigen was identified, notable progress has been made on exploration how florigen gene expression is genetically controlled. In Arabidopsis expression of certain key flowering integrators such as FLOWERING LOCUS C (FLC) and FLOWERING LOCUS T (FT) are also epigenetically regulated by various chromatin modifications, however, very little is known in rice on this aspect until very recently. This review summarized the advances of both genetic networks and chromatin modifications in rice flowering time control, attempting to give a complete view of the genetic and epigenetic architecture in complex network of rice flowering pathways.     

Specification of Arabidopsis floral meristem identity by repression of flowering time genes

Flowering plants produce floral meristems in response to intrinsic and extrinsic flowering inductive signals. In Arabidopsis, the floral meristem identity genes LEAFY (LFY) and APETALA1 (AP1) are activated to play a pivotal role in specifying floral meristems during floral transition. We show here that the emerging floral meristems require AP1 to partly specify their floral identities by directly repressing a group of flowering time genes, including SHORT VEGETATIVE PHASE (SVP), AGAMOUS-LIKE 24 (AGL24) and SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1). In wild-type plants, these flowering time genes are normally downregulated in emerging floral meristems. In the absence of AP1, these genes are ectopically expressed, transforming floral meristems into shoot meristems. By post-translational activation of an AP1-GR fusion protein and chromatin immunoprecipitation assays, we further demonstrate the repression of these flowering time genes by induced AP1 activity and in vivo AP1 binding to the cis-regulatory regions of these genes. These findings indicate that once AP1 is activated during the floral transition, it acts partly as a master repressor in floral meristems by directly suppressing the expression of flowering time genes, thus preventing the continuation of the shoot developmental program.     

Differential expression of genes important for adaptation in Capsella bursa-pastoris (Brassicaceae)

Understanding the genetic basis of natural variation is of primary interest for evolutionary studies of adaptation. In Capsella bursa-pastoris, a close relative of Arabidopsis (Arabidopsis thaliana), variation in flowering time is correlated with latitude, suggestive of an adaptation to photoperiod. To identify pathways regulating natural flowering time variation in C. bursa-pastoris, we have studied gene expression differences between two pairs of early- and late-flowering C. bursa-pastoris accessions and compared their response to vernalization. Using Arabidopsis microarrays, we found a large number of significant differences in gene expression between flowering ecotypes. The key flowering time gene FLOWERING LOCUS C (FLC) was not differentially expressed prior to vernalization. This result is in contrast to those in Arabidopsis, where most natural flowering time variation acts through FLC. However, the gibberellin and photoperiodic flowering pathways were significantly enriched for gene expression differences between early- and late-flowering C. bursa-pastoris. Gibberellin biosynthesis genes were down-regulated in late-flowering accessions, whereas circadian core genes in the photoperiodic pathway were differentially expressed between early- and late-flowering accessions. Detailed time-series experiments clearly demonstrated that the diurnal rhythm of CIRCADIAN CLOCK-ASSOCIATED1 (CCA1) and TIMING OF CAB EXPRESSION1 (TOC1) expression differed between flowering ecotypes, both under constant light and long-day conditions. Differential expression of flowering time genes was biologically validated in an independent pair of flowering ecotypes, suggesting a shared genetic basis or parallel evolution of similar regulatory differences. We conclude that genes involved in regulation of the circadian clock, such as CCA1 and TOC1, are strong candidates for the evolution of adaptive flowering time variation in C. bursa-pastoris.     

西红花FT同源基因的表达及功能分析

FT（FLOWERING LOCUS T）及其同源基因作为三大开花途径整合子之一,被认为是调控植物开花的重要基因。为了深入研究FT同源基因的功能以及西红花（Crocus sativus L.）开花的分子机理,对已报道的3个西红花FT同源基因（CsatFT1、CsatFT2和CsatFT3）进行分离及分析。gDNA包含长度分别为835、1 642和1 132 bp的完整开放阅读框（ORF）,均具有4个外显子和3个内含子;cDNA包含长度分别为528、525和540 bp的ORF,分别编码175、174和179个氨基酸;系统进化分析表明,CsatFT1、CsatFT2、CsatFT3分别和同为单子叶植物的水仙（Narcissus chinensis）NtFT、麝香百合（Lilium longiflorum）LlFT和洋葱（Allium cepa）AcFT1表现出较近的遗传距离。qRT-PCR分析结果显示,小球茎膨大阶段前期,CsatFT1、CsatFT2、CsatFT3在叶片中表达水平最高,侧根中次之,子球茎、主根中极低几乎检测不到;小球茎膨大阶段后期,CsatFT1、CsatFT2、CsatFT3都在子球茎中表达水平较高,在顶芽中几乎检测不到;室内储藏开花阶段,CsatFT1、CsatFT2、CsatFT3在柱头中表达水平最高,叶中次之,花瓣和花药中较低几乎检测不到。通过观测转基因烟草（Nicotiana tabacum）和转基因拟南芥（Arabidopsis thaliana）植株表型发现,CsatFT1,CsatFT2和CsatFT3均具有促进植物提早开花的功能。     

Adaptation to climate through flowering phenology: a case study in Medicago truncatula

Local climatic conditions likely constitute an important selective pressure on genes underlying important fitness‐related traits such as flowering time, and in many species, flowering phenology and climatic gradients strongly covary. To test whether climate shapes the genetic variation on flowering time genes and to identify candidate flowering genes involved in the adaptation to environmental heterogeneity, we used a large Medicago truncatula core collection to examine the association between nucleotide polymorphisms at 224 candidate genes and both climate variables and flowering phenotypes. Unlike genome‐wide studies, candidate gene approaches are expected to enrich for the number of meaningful trait associations because they specifically target genes that are known to affect the trait of interest. We found that flowering time mediates adaptation to climatic conditions mainly by variation at genes located upstream in the flowering pathways, close to the environmental stimuli. Variables related to the annual precipitation regime reflected selective constraints on flowering time genes better than the other variables tested (temperature, altitude, latitude or longitude). By comparing phenotype and climate associations, we identified 12 flowering genes as the most promising candidates responsible for phenological adaptation to climate. Four of these genes were located in the known flowering time QTL region on chromosome 7. However, climate and flowering associations also highlighted largely distinct gene sets, suggesting different genetic architectures for adaptation to climate and flowering onset.     

Flowering time genes Heading date 1 and Early heading date 1 together control panicle development in rice

Although flowering time is often associated with plant size, little is known about how flowering time genes affect plant architecture. We grew four rice lines having different flowering time genotypes (hd1 ehd1, hd1 Ehd1, Hd1 ehd1 and Hd1 Ehd1) under distinct photoperiod conditions. By using genotype-treatment combinations that resulted in similar flowering times, we were able to compare the effects of flowering time genes on traits related to plant architecture. The results revealed that the combination of Heading-date 1 (Hd1) and Early heading date 1 (Ehd1) can reduce the number of primary branches in a panicle, resulting in smaller spikelet numbers per panicle; this occurs independently of the control of flowering time. In addition, expression of the Hd3a and Rice Flowering-locus T 1 (RFT1) florigen genes was up-regulated in leaves of the Hd1 Ehd1 line at the time of the floral transition. We further revealed that Hd1 and/or Ehd1 caused up-regulation of Terminal Flower 1-like genes and precocious expression of panicle formation-related genes at shoot apical meristems during panicle development. Therefore, two key flowering time genes, Hd1 and Ehd1, can control panicle development in rice; this may affect crop yields in the field through florigen expression in leaf.