The role of cryptochrome 2 in flowering in Arabidopsis

We have investigated the genetic interactions between cry2 and the various flowering pathways in relation to the regulation of flowering by photoperiod and vernalization. For this, we combined three alleles of CRY2, the wild-type CRY2-Landsberg erecta (Ler), a cry2 loss-of-function null allele, and the gain-of-function CRY2-Cape Verde Islands (Cvi), with mutants representing the various photoreceptors and flowering pathways. The analysis of CRY2 alleles combined with photoreceptor mutants showed that CRY2-Cvi could compensate the loss of phyA and cry1, also indicating that cry2 does not require functional phyA or cry1. The analysis of mutants of the photoperiod pathway showed epistasis of co and gi to the CRY2 alleles, indicating that cry2 needs the product of CO and GI genes to promote flowering. All double mutants of this pathway showed a photoperiod response very much reduced compared with Ler. In contrast, mutations in the autonomous pathway genes were additive to the CRY2 alleles, partially overcoming the effects of CRY2-Cvi and restoring day length responsiveness. The three CRY2 alleles were day length sensitive when combined with FRI-Sf2 and/or FLC-Sf2 genes, which could be reverted when the delay of flowering caused by FRI-Sf2 and FLC-Sf2 alleles was removed by vernalization. In addition, we looked at the expression of FLC and CRY2 genes and showed that CRY2 is negatively regulated by FLC. These results indicate an interaction between the photoperiod and the FLC-dependent pathways upstream to the common downstream targets of both pathways, SOC1 and FT.     

A role of flowering genes in the tolerance of Arabidopsis thaliana to cucumber mosaic virus

The genetic basis of plant tolerance to parasites is poorly understood. We have previously shown that tolerance of Arabidopsis thaliana to its pathogen cucumber mosaic virus is achieved through changes in host life-history traits on infection that result in delaying flowering and reallocating resources from vegetative growth to reproduction. In this system we analyse here genetic determinants of tolerance using a recombinant inbred line family derived from a cross of two accessions with extreme phenotypes. Three major quantitative trait loci for tolerance were identified, which co-located with three flowering repressor genes, FLC, FRI, and HUA2. The role of these genes in tolerance was further examined in genotypes carrying functional or nonfunctional alleles. Functional alleles of FLC together with FRI and/or HUA2 were required for both tolerance and resource reallocation from growth to reproduction. Analyses of FLC alleles from wild accessions that differentially modulate flowering time showed that they ranked differently for their effects on tolerance and flowering. These results pinpoint a role of FLC in A. thaliana tolerance to cucmber mosaic virus, which is a novel major finding, as FLC has not been recognized previously to be involved in plant defence. Although tolerance is associated with a delay in flowering that allows resource reallocation, our results indicate that FLC regulates tolerance and flowering initiation by different mechanisms. Thus, we open a new avenue of research on the interplay between defence and development in plants.     

early bolting in short days: an Arabidopsis mutation that causes early flowering and partially suppresses the floral phenotype of leafy

The time of flowering in Arabidopsis is controlled by multiple endogenous and environmental signals. Some of these signals promote the onset of flowering, whereas others repress it. We describe here the isolation and characterization of two allelic mutations that cause early flowering and define a new locus, EARLY BOLTING IN SHORT DAYS (EBS). Acceleration of flowering time in the ebs mutants is especially conspicuous under short-day photoperiods and results from a reduction of the adult vegetative phase of the plants. In addition to the early flowering phenotype, ebs mutants show a reduction in seed dormancy, plant size, and fertility. Double mutant analysis with gibberellin-deficient mutants indicates that both the early-flowering and the precocious-germination phenotypes require gibberellin biosynthesis. Analysis of the genetic interactions among ebs and several mutations causing late flowering shows that the ft mutant phenotype is epistatic over the early flowering of ebs mutants, suggesting that the precocious flowering of ebs requires the FT gene product. Finally, the ebs mutation causes an increase in the level of expression of the floral homeotic genes APETALA3 (AP3), PISTILLATA (PI), and AGAMOUS (AG) and partially rescues the mutant floral phenotype of leafy-6 (lfy-6) mutants. These results suggest that EBS participates as a negative regulator in developmental processes such as germination, flowering induction, and flower organ specification.     

变温调控对苦苣温度敏感期及其抽薹开花的影响

将苦苣幼苗在温室暖环境(17℃)和冷环境(平均10.5℃)下每周进行双向互换,研究了苦苣对温度的敏感时期及其在不同温度条件下的生长发育规律.结果表明:苦苣幼苗在暖环境和冷环境下的生长速度差异显著;在2～17℃的设定温度范围内,苦苣没有一个明显的抽薹温度敏感时期,均能感受外界温度从而进行春化诱导,且低温有利于花芽的分化.苦苣在暖环境下和冷环境下均能完成花芽分化和抽薹,冷环境下花芽分化的时间为24 d,暖环境下为60 d.在暖环境下苦苣抽薹比冷环境下抽薹晚7d,暖环境和冷环境下苦苣从播种到抽薹的平均时间分别为111和104 d;开花诱导完成后从暖环境转到冷环境下,苦苣的抽薹速率明显高于从冷环境转到暖环境,在这段时间环境温度越高,抽薹速率越快.     

Characterization of the spermidine synthase-related gene family in Arabidopsis thaliana

The Arabidopsis genome contains four genes that encode proteins similar to both spermidine synthase and spermine synthase of other organisms. Our previous study revealed that one of these genes, designated ACAULIS5 (ACL5), encodes spermine synthase and that its null mutation results in a severe defect in the elongation of stem internodes. Here we report the characterization of the other three genes, designated SPDS1, SPDS2 and SPDS3. Our results showed that SPDS1 and SPDS2 possess spermidine synthase activity in yeast spermidine synthase-deficient mutants, but the enzyme activity of SPDS3 remained to be determined. RNA gel blot analysis revealed that all of these genes are expressed in all plant organs but show different responses to exogenous plant hormones, suggesting that they are involved in different aspects of growth by modulating the contents of polyamines in plant cells.     

FLC: A key regulator of flowering time in Arabidopsis

The timing of floral transition has significant consequences for reproductive success in plants. The molecular genetic dissection of flowering time control in Arabidopsis identified an integrated network of pathways that quantitatively control this developmental switch. A central player in this process is the FLOWERING LOCUS C gene (FLC), which blocks flowering by inhibiting the genes required to switch the meristem from vegetative to floral development. Three systems (the FRIGIDA gene, vernalization, and the autonomous pathway) all influence the state of FLC. Last years many new genes have been identified that regulate FLC expression, and most of them are involved in the modification of FLC chromatin. This review focuses on recent insights in FLC regulation.     

Cadmium induces early flowering in Arabidopsis

We found that cadmium promoted flowering in Arabidopsis and suppressed nitric oxide accumulation in leaves. Supplementation with NO donor SNP delayed flowering, whereas application of NO scavenger cPTIO further promoted the transition from vegetative to reproductive stage under Cd stress. Semi-quantitative RT-PCR showed that Cd treatment up-regulated the expression of CONSTANS and FLOWERING LOCUS T, whereas down-regulated the expression of FLOWERING LOCUS C.     

Analysis of the role of the late-flowering locus,GI, in the flowering of Arabidopsis thaliana

The mutation gigantea (gi) is recessive and belongs to the late-flowering mutations in Arabidopsis thaliana. The late-flowering mutations result in a pronounced delay in flowering due to a prolonged phase of vegetative growth, which is manifested by an increased number of primary foliage leaves in the rosette (i.e. vegetative nodes). To examine the nature of the gi mutation, detailed phenotypic analysis was carried out for three representative mutant alleles. The results indicate that gi mutants have a defect in the promotion of the floral induction process by long-day photoperiods and not in the flowering process per se. Temperature-shift experiments using a partially conditional allele were employed to determine the timing of the functional requirement for the product of the GI locus. The end of the deduced functional period corresponds to the period at which transition of the apical meristem from the vegetative to the reproductive phase occurs. Such timing is in good agreement with the postulated role of the GI locus. These results demonstrate that the GI locus is involved in the promotion of floral initiation (entrance of the meristem into the transitional stage) by long-day photoperiods.     

A gain-of-function mutation of Arabidopsis cryptochrome1 promotes flowering

Plants use different classes of photoreceptors to collect information about their light environment. Cryptochromes are blue light photoreceptors that control deetiolation, entrain the circadian clock, and are involved in flowering time control. Here, we describe the cry1-L407F allele of Arabidopsis (Arabidopsis thaliana), which encodes a hypersensitive cryptochrome1 (cry1) protein. Plants carrying the cry1-L407F point mutation have elevated expression of CONSTANS and FLOWERING LOCUS T under short-day conditions, leading to very early flowering. These results demonstrate that not only the well-studied cry2, with an unequivocal role in flowering promotion, but also cry1 can function as an activator of the floral transition. The cry1-L407F mutants are also hypersensitive toward blue, red, and far-red light in hypocotyl growth inhibition. In addition, cry1-L407F seeds are hypersensitive to germination-inducing red light pulses, but the far-red reversibility of this response is not compromised. This demonstrates that the cry1-L407F photoreceptor can increase the sensitivity of phytochrome signaling cascades. Molecular dynamics simulation of wild-type and mutant cry1 proteins indicated that the L407F mutation considerably reduces the structural flexibility of two solvent-exposed regions of the protein, suggesting that the hypersensitivity might result from a reduced entropic penalty of binding events during downstream signal transduction. Other nonmutually exclusive potential reasons for the cry1-L407F gain of function are the location of phenylalanine-407 close to three conserved tryptophans, which could change cry1's photochemical properties, and stabilization of ATP binding, which could extend the lifetime of the signaling state of cry1.     

Day-length perception and the photoperiodic regulation of flowering in Arabidopsis

The flowering of Arabidopsis plants is accelerated by long-day photoperiods, and recent genetic studies have identified elements of the photoperiodic timing mechanism. These elements comprise genes that regulate the function of the circadian clock, photoreceptors, and downstream components of light signaling pathways. These results provide evidence for the role of the circadian clock in photoperiodic time measurement and suggest that photoperiod perception may follow Pittendrigh's external coincidence model. T-cycle experiments indicated that changes in the timing of circadian rhythms, relative to dawn and dusk, correlated with altered flowering time. Thus, the perception of photoperiod maybe mediated by adjustments in the phase of the circadian cycle that arise upon re-entrainment to a different light-dark cycle. The nature of the rhythm underlying the floral response is not known, but candidate molecules have been identified.     

A MADS domain gene involved in the transition to flowering in Arabidopsis

Flowering time in many plants is triggered by environmental factors that lead to uniform flowering in plant populations, ensuring higher reproductive success. So far, several genes have been identified that are involved in flowering time control. AGL20 (AGAMOUS LIKE 20) is a MADS domain gene from Arabidopsis that is activated in shoot apical meristems during the transition to flowering. By transposon tagging we have identified late flowering agl20 mutants, showing that AGL20 is involved in flowering time control. In previously described late flowering mutants of the long-day and constitutive pathways of floral induction the expression of AGL20 is down-regulated, demonstrating that AGL20 acts downstream to the mutated genes. Moreover, we can show that AGL20 is also regulated by the gibberellin (GA) pathway, indicating that AGL20 integrates signals of different pathways of floral induction and might be a central component for the induction of flowering. In addition, the constitutive expression of AGL20 in Arabidopsis is sufficient for photoperiod independent flowering and the over-expression of the orthologous gene from mustard, MADSA, in the classical short-day tobacco Maryland Mammoth bypasses the strict photoperiodic control of flowering.     

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.     

Effects of antimetabolites on flowering in Arabidopsis

Effects of five kinds of antimetabolites on flowering in Arabidopsiswere studied. 8-Azaguanine, acriflavine, ethidium bromide andstreptomycin accelerated flowering but cycloheximide did not.The accelerating effect of 8-azaguanine was reversed by guanine. (Received April 30, 1971; )     

cDNA-AFLP analysis on bolting or flowering of flowering Chinese cabbage and molecular characteristics of BrcuDFR-like/BrcuAXS gene

The molecular basis of flower bud differentiation in flowering Chinese cabbage (Brassica rapa L. ssp. Chinensis var. utilis Tsen et Lee) was studied in this work. Samples were taken from two varieties, the early-blooming "Youqin 49" and the late-blooming "Youqingtiancaixin 80", at five different developmental stages and studied via cDNA-AFLP. Nineteen expression sequence tags (ESTs) associated with bolting or flowering were isolated and cloned. Blast results indicated that 15 ESTs were involved in the synthesis of anthocayanins, photosynthesis, signal transduction, and phytochrome production. Two ESTs had high similarity to hypothetical proteins with unknown function. Two other ESTs shared no similarity to any sequence in the NCBI database and potentially may be newly identified genes. The deduced amino acid sequences of EST amplified by primer A6T4 or A8T4 had high similarity to both dihydroflavonol reductase (DFR) and UDP-D: -apiose/UDP-D: -xylose synthase (AXS), thus was named BrcuDFR-like/BrcuAXS. Using the cDNA sequence, a putative BrcuDFR-like/BrcuAXS gene was cloned and characterized from flowering Chinese cabbage via rapid amplification of cDNA ends (RACE). The full-length cDNA has 1332?bp with an open frame of 919?bp which codes for a polypeptide of 313 amino acids. The corresponding genome sequence is 2,046?bp. Comparison of cDNA and its corresponding genomic sequence indicates that BrcuDFR-like/BrcuAXS contains 9 exons and 8 introns. The temporal expression patterns indicated the gene is more likely to encode the DFR protein, which catalyzes the synthesis of anthocayanins, than UDP-D: -apiose/UDP-D: -xylose synthase (AXS), which catalyzes the conversion of UDP-D: -glucuronate to a mixture of UDP-D: -apiose and UDP-D: -xylose. Further work is needed to determine what role BrcuDFR-like/BrcuAXS plays during floral organ development.