Integration of flowering signals in winter-annual Arabidopsis

Photoperiod is the primary environmental factor affecting flowering time in rapid-cycling accessions of Arabidopsis (Arabidopsis thaliana). Winter-annual Arabidopsis, in contrast, have both a photoperiod and a vernalization requirement for rapid flowering. In winter annuals, high levels of the floral inhibitor FLC (FLOWERING LOCUS C) suppress flowering prior to vernalization. FLC acts to delay flowering, in part, by suppressing expression of the floral promoter SOC1 (SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1). Vernalization leads to a permanent epigenetic suppression of FLC. To investigate how winter-annual accessions integrate signals from the photoperiod and vernalization pathways, we have examined activation-tagged alleles of FT and the FT homolog, TSF (TWIN SISTER OF FT), in a winter-annual background. Activation of FT or TSF strongly suppresses the FLC-mediated late-flowering phenotype of winter annuals; however, FT and TSF overexpression does not affect FLC mRNA levels. Rather, FT and TSF bypass the block to flowering created by FLC by activating SOC1 expression. We have also found that FLC acts as a dosage-dependent inhibitor of FT expression. Thus, the integration of flowering signals from the photoperiod and vernalization pathways occurs, at least in part, through the regulation of FT, TSF, and SOC1.     

FLC基因表达在植物春化过程中的作用

在对以往有关不同开花途径研究简要总结的基础上综述了FLC基因在春化过程中的作用。近期以拟南芥不同生态型和突变体为模式的研究结果表明基因FLC可能是春化反应的关键基因。研究发现 ,FLC的表达水平与植株低温处理的时间呈数量关系 ,低温处理时间越长 ,FLC的表达越弱 ,去甲基化也可能对FLC起负调控的作用。同时FLC也存在于自主开花途径中 ,与其他基因共同作用以调节植株开花时间。而FLC的表达对开花起抑制作用。一系列研究表明 ,春化的低温作用可能在于相关基因的去甲基化 ,消除了FLC对开花的抑制作用 ,从而解除赤霉素合成途径的封锁最终导致植株在一定时期开花。     

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.     

FLC基因表达在植物春化过程中的作用

在对以往有关不同开花途径研究简要总结的基础上综述了FLC基因在春化过程中 的作用。近期以拟南芥不同生态型和突变体为模式的研究结果表明基因FLC可能是春化反应的关键基因。研究发现,FLC的表达水平与植株低温处理的时间呈数量关系,低温处理时间越长,FLC的表达越弱,去甲基化也可能对FLC起负调控的作用。同时FLC也存在于自主开花途径中,与其他基因共同作用以调节植株开花时间。而FLC的表达对开花起抑制作用。一系列研究表明,春化的低温作用可能在于相关基因的去甲基化,消除了FLC对开花的抑制作用,从而解除赤霉素合成途径的封锁最终导致植株在一定时期开花。     

Quantitative effects of vernalization on FLC and SOC1 expression

Prolonged exposure to cold results in early flowering in Arabidopsis winter annual ecotypes, with longer exposures resulting in a greater promotion of flowering than shorter exposures. The promotion of flowering is mediated through an epigenetic down-regulation of the floral repressor FLOWERING LOCUS C (FLC). We present results that provide an insight into the quantitative regulation of FLC by vernalization. Analysis of the effect of seed or plant cold treatment on FLC expression indicates that the time-dependent nature of vernalization on FLC expression is mediated through the extent of the initial repression of FLC and not by affecting the ability to maintain the repressed state. In the over-expression mutant flc-11, the time-dependent repression of FLC correlates with the proportional deacetylation of histone H3. Our results indicate that sequences within intron 1 and the activities of both VERNALIZATION1 (VRN1) and VERNALIZATION2 (VRN2) are required for efficient establishment of FLC repression; however, VRN1 and VRN2 are not required for maintenance of the repressed state during growth after the cold exposure. SUPPRESSOR OF OVER-EXPRESSION OF CO 1 (SOC1), a downstream target of FLC, is quantitatively induced by vernalization in a reciprocal manner to FLC. In addition, we show that SOC1 undergoes an acute induction by both short and long cold exposures.     

SHB1 plays dual roles in photoperiodic and autonomous flowering

Flowering was initiated by the integration of environmental signals such as day-length with the internal development status in Arabidopsis, a facultative long-day plant. The photoperiodic flowering involves two key components, CONSTANS and FT, whereas the autonomous flowering is operated through a central quantitative floral repressor, FLC, and several other genes that act upstream of FLC. SOC1 acts downstream to integrate the flowering signals from the two pathways. Here, we report that SHB1 plays dual roles in both photoperiodic and autonomous flowering. shb1-D, a gain-of-function mutant, flowered early and shb1, a loss-of-function allele, flowered late under both long days and short days. The shb1-D mutation activated the expression of CO, FT, and SOC1 under both long and short days, and however, the co-2 mutation attenuated the shb1-D activated expression of FT and SOC1 only under long days but not short days. The shb1-D or shb1 mutations also reduced and increased, respectively, the expression of FLC under both long and short days. Transgenic remedy of FLC to wide-type level in shb1-D background also reverted shb1-D flowering and FT or SOC1 expression to wild type mostly under short days. Furthermore, the shb1-D suppression on FLC expression is likely operated through LD as ld-3 blocked this suppression and SHB1 appears to act upstream of LD. In summary, SHB1 represents signaling steps that regulate CO expression in leaves and LD or FLC expression in either leaves or shoot apical meristem, contributing to a threshold expression of SOC1 in shoot apical meristem for floral initiation.     

拟南芥开花时间调控的研究进展

调控开花时间是大多数植物由营养生长向生殖生长转化的一个重要生长发育过程.影响拟南芥开花时间的因素有很多,其中光照和温度是两个主要的外部因素,而赤霉素(GA)和一些自主性因子是主要的内部因素.目前,一般按照对以上因素的反应将晚花突变体归于四条开花调控途径:光周期途径、春化途径、自主途径和GA途径.在不断变化的外部环境条件和内部生理条件下,这些途径通过一些主要的整合基因如SOC1、FT、LFY等实现了对拟南芥开花时间的精确调控.     

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.     

Analysis of Flowering Time Control in Arabidopsis by Comparison of Double and Triple Mutants

Three genetic pathways promote flowering of Arabidopsis under long photoperiods. These pathways are represented by the genes CO, FCA, and GA1, which act in the long-day, autonomous, and gibberellin pathways, respectively. To test whether these are the only pathways that promote flowering under long photoperiods, the co-2 fca-1 ga1-3 triple mutant was constructed. These plants never flowered under long- or short-day conditions, indicating that the three pathways impaired by these mutations are absolutely required for flowering under these conditions. The triple mutant background represents a "vegetative ground state" enabling the roles of single pathways to be described in the corresponding double mutants. The phenotypes of plants carrying all eight combinations of wild-type and mutant alleles at the three loci were compared under long- and short-day conditions. This analysis demonstrated that under long photoperiods the long-day pathway promoted flowering most effectively, whereas under short photoperiods the gibberellin pathway had the strongest effect. The autonomous pathway had a weak effect when acting alone under either photoperiod but appeared to play an important role in facilitating the promotion of flowering by the other two pathways. The vegetative phenotype of the triple mutant could be overcome by vernalization, suggesting that a fourth pathway promoted flowering under these conditions. These observations are discussed in light of current models describing the regulation of flowering time in Arabidopsis.