2. The role of plant growth regulators in promotion of flowering

Research on flowering in forest trees has pursued two main objectives: 1) developing methods for stimulating flowering of selected genotypes to accelerate progeny testing, and 2) hastening the production of genetically-improved seeds in seed orchards. Cultural techniques have been used to some extent in the past for these purposes, but the use of plant-growth regulators (PGRs), particularly the gibberellins (GAs), has been increasingly successful. GAs have been shown to induce flowering in most conifers, but GA application on an operational basis has been worked out for only a few species. GAs have been applied effectively by topical treatment, by stem or branch injections, and by foliar sprays. The timing and concentration of applied chemicals are important and vary from species to species. Another PGR, naphthalene-acetic acid, is synergistic with GAs in some species. Adjunct cultural treatments are often used in combination with treatment with GAs. The success of treatment with GAs is determined to some extent upon the stage of development of the treated plants. The endogenous PGRs regulating flowering are not completely known but it appears that the GAs, the cytokinins, and bascisic acid may all be changed by treatments which induce flowering. Improved techniques becoming available for measuring PGRs are beginning to stimulate research on the endogenous PGRs. The PGRs will continue to play a major role in stimulating flowering in forest trees.     

Does CK2 affect flowering time by modulating the autonomous pathway in Arabidopsis?

CK2 (Casein kinase II), a ubiquitous Ser/Thr kinase, affects multiple developmental and stress response pathways in Arabidopsis, including flowering time under both long- and short-day conditions through the photoperiod and autonomous pathways. CK2 phosphorylates central clock components, CCA1 and LHY, to modulate circadian clock that regulates flowering time through the photoperiod pathway. However, how CK2 regulates flowering time through the autonomous pathway is still unknown. Analyses of phosphorylation sites using several prediction softwares show that most of the autonomous pathway components have multiple CK2 phosphorylation sites. CK2 might phosphorylate any or all of these components to modulate their activity/stability resulting in altered expression of FLC that drives flowering time through the autonomous pathway.     

The impact of photoperiod insensitive Ppd-1a mutations on the photoperiod pathway across the three genomes of hexaploid wheat (Triticum aestivum)

Flowering time is a trait that has been extensively altered during wheat domestication, enabling it to be highly productive in diverse environments and providing a rich source of variation for studying adaptation mechanisms. Hexaploid wheat is ancestrally a long-day plant, but many environments require varieties with photoperiod insensitivity (PI) that can flower in short days. PI results from mutations in the Ppd-1 gene on the A, B or D genomes, with individual mutations conferring different degrees of earliness. The basis of this is poorly understood. Using a common genetic background, the effects of A, B and D genome PI mutations on genes of the circadian clock and photoperiod pathway were studied using genome-specific expression assays. Ppd-1 PI mutations did not affect the clock or immediate clock outputs, but affected TaCO1 and TaFT1, with a reduction in TaCO1 expression as TaFT1 expression increased. Therefore, although Ppd-1 is related to PRR genes of the Arabidopsis circadian clock, Ppd-1 affects flowering by an alternative route, most likely by upregulating TaFT1 with a feedback effect that reduces TaCO1 expression. Individual genes in the circadian clock and photoperiod pathway were predominantly expressed from one genome, and there was no genome specificity in Ppd-1 action. Lines combining PI mutations on two or three genomes had enhanced earliness with higher levels, but not earlier induction, of TaFT1, showing that there is a direct quantitative relationship between Ppd-1 mutations, TaFT1 expression and flowering.     

The Interaction of Gibberellins and Photoperiod in the Control of Potato Tuberization

Solanum tuberosum ssp. andigena plants require a short-day (SD) photoperiod for tuber formation, a process that is also affected by gibberellins (GAs). Grafting experiments have confirmed that the photoperiod is perceived in the leaves. Tuber formation, however, usually takes place in the underground stolons. In this review, photoperiod-dependent tuberization has been divided into five chronological events: SD photoperiod perception, short-term adaptive responses to SD conditions, generation and transport of tuber-inducing signal(s), tuber formation, and long-term adaptive responses to tuber growth. Within this frame of study, the interaction of GAs and photoperiod is revised. Similar to the flowering process in Arabidopsis, we suggest the existence of two independent pathways that control tuber formation: a photoperiod-dependent pathway and a GA-dependent pathway. Nevertheless, photoperiod-dependent tuber formation requires the action of GAs at specific stages to orchestrate this complex process of development.     

Effects of Interactions between Low-temperature Treatments, Gibberellin (GA3) and Photoperiod on Flowering and Stem Height of Spring Rape (Brassica napus var. annua)

Exogenous gibberellin A₃(GA₃) reduced the number of leaf nodesat flowering and time to flowering and increased the stem heightat flowering in three genotypes of spring rape (Brassica napusvar.annua L.). The responses to GA₃were similar to those forlong days (LD) and low-temperature treatments, suggesting thatthe effect of photoperiod and the vernalization response areprobably mediated through gibberellins. The response to exogenousGA₃was greatest in non-cold-treated plants in short days (SD)suggesting that endogenous GAs are limiting in these conditions.CCC, an inhibitor of gibberellin biosynthesis, caused a smallincrease in the number of leaf nodes at flowering and time toflowering and a small decrease in the stem height at flowering,but unexpectedly, its effect was hardly influenced by the applicationof exogenous GA₃. Genotypes that showed the clearest responsesto the treatments with regard to the number of leaf nodes atflowering and time to flowering did not show the clearest responseswith regard to the stem height at flowering; the pattern ofresponses of the number of leaf nodes at flowering and timeto flowering was distinct from that of stem height at flowering.This indicates that flower formation and stem elongation areseparable developmental processes which may be controlled bydifferent endogenous gibberellins, different levels of a specificendogenous gibberellin, or different responses to gibberellin.Copyright 1999 Annals of Botany Company Brassica napus var. annua, gibberellin, photoperiod, spring rape, vernalization.     

Quantitation of gibberellins and indoleacetic acid in Begonia leaves: Relationship with environment, regeneration and flowering

Indole-3-acetic acid (IAA) was purified by high performance liquid chromatography (HPLC) and identified by gas chromatography - mass spectrometry (GC-MS) in leaf extracts of Begonia × cheimantha Everett cv. Nova. The content of IAA and of gibberellins A4, A9, A19 and A20 (GAs) previously identified in this material, were quantified by GC-MS in leaves of Begonia plants grown under different temperature and daylength conditions, using deuterated compounds as internal standards. GA1, which was also identified, was present in too low quantities for reliable quantitation. Rapid and significant decreases (within 2–4 days) occurred in the content of both IAA and GAs when the plants were transferred from conditions which are non-inductive for adventitious bud formation and flowering (24°C/long day) to inductive conditions (24°C/short day, 15°C/long day or 15°C/short day). GA4 and GA9 were affected by photoperiod only, whereas IAA, GA19 and GA20 were affected by both photoperiod and temperature. The data suggest that biosynthesis of GA9 and GA4 are blocked in short days at a step located before GA9. Conversion of GA19 to GA20 seemed to be blocked by both short days and low temperature, while an additional block located before GA19 seemed to be imposed in 15°C/short day. The results confirm earlier results and support the hypothesis that photoperiod and temperature effects in Begonia are mediated by endogenous hormones.     

Diversified mechanisms for regulating flowering time in a short-day plant rice

Flowering in rice is influenced by not only endogenous factors that comprise an autonomous pathway, but also environmental effects, such as photoperiod, water availability, and temperature just before floral initiation. Recent molecular genetics studies have elucidated the functional roles of genes involved in the photoperiod pathway, e.g., photoreceptors, circadian clock components, and short-day (SD) promotion factors. Although these molecular players are well conserved between rice andArabidopsis, their actual genetic functions are distinct. This is exemplified byHd1 (aCO counterpart) and phytochromes, in particular, ricePHYA. Hd1 has a dual role in regulating flowering time and the expression ofHd3a (anFT counterpart) repression under long-day (LD) conditions while promotion under SDs. Models have been proposed to explain these photoperiod-dependent antagonistic activities. Some regulatory factors are present in only one of the model systems, e.g.,FLC inArabidopsis orEhd1 in rice. Furthermore, epistatic relationships vary among such flowering regulators asHd3a (FT), OsMADS50 (SOCT), andOsMADS14 (AP1). Further experiments to probe these differences will be essential to enlarging our understanding of the diversified flowering regulation mechanisms in rice.     

Photoperiod sensing of the circadian clock is controlled by EARLY FLOWERING 3 and GIGANTEA

ELF3 and GI are two important components of the Arabidopsis circadian clock. They are not only essential for the oscillator function but are also pivotal in mediating light inputs to the oscillator. Lack of either results in a defective oscillator causing severely compromised output pathways, such as photoperiodic flowering and hypocotyl elongation. Although single loss of function mutants of ELF3 and GI have been well studied, their genetic interaction remains unclear. We generated an elf3 gi double mutant to study their genetic relationship in clock‐controlled growth and phase transition phenotypes. We found that ELF3 and GI repress growth differentially during the night and the day, respectively. Circadian clock assays revealed that ELF3 and GI are essential that enable the oscillator to synchronize the endogenous cellular mechanisms to external environmental signals. In their absence, the circadian oscillator fails to synchronize to the light–dark cycles even under diurnal conditions. Consequently, clock‐mediated photoperiod‐responsive growth and development are completely lost in plants lacking both genes, suggesting that ELF3 and GI together convey photoperiod sensing to the central oscillator. Since ELF3 and GI are conserved across flowering plants and represent important breeding and domestication targets, our data highlight the possibility of developing photoperiod‐insensitive crops by adjusting the allelic combination of these two key genes.     

Gibberellins in apical shoot meristems of flowering and vegetative sugarcane

Gibberellins A₁, A₃, iso-A₃, A₄, A₁₉, A₂₀, and A₃₆ were identified by gas chromatography-selected ion monitoring in apices of sugarcane (Saccharum spp. hybrids). Flowering apices (i.e., 2–4 cm panicle) contained 8–9 times more (estimated by bioassay) endogenous gibberellins A and iso-GA₃ (ratio of 1:6:8, respectively; in total 51 ng g^–1 fresh weight) than vegetative apices (6.4 ng g^–1 fresh weight). Vegetative apices contained small but significant levels of GA₁₉, which could not be detected in flowering apices; vegetative apices also contained approximately four times more of a GA₃₆-like substance than flowering apices. Since the two apex types developed under the same photoperiod, the increased levels of GA and iso-GA₃ and the reduced levels of GA₁₉ and GA₃₆-like substances are correlated with the flowering state rather than with photoperiod or photoperiod changes per se. Since there were relatively high levels of C₁₉ GAs along with low levels of C₂₀ GAs in flowering apices, and since the converse is true in vegetative apices, metabolism of C₂₀ to C₁₉ GAs may be enhanced in flowering apices.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Flowering of the grass Lolium perenne: effects of vernalization and long days on gibberellin biosynthesis and signaling

Almost 50 years ago, it was shown that gibberellin (GA) applications caused flowering in species normally responding to cold (vernalization) and long day (LD). The implication that GAs are involved with vernalization and LD responses is examined here with the grass Lolium perenne. This species has an obligatory requirement for exposure to both vernalization and LD for its flowering (inflorescence initiation). Specific effects of vernalization or LD on GA synthesis, content, and action have been documented using four treatment pairs: nonvernalized or vernalized plants exposed to short days (SDs) or LDs. Irrespective of vernalization status, exposure to two LDs increased expression of L. perenne GA 20-oxidase-1 (LpGA20ox1), a critical GA biosynthetic gene, with endogenous GAs increasing by up to 5-fold in leaf and shoot. In parallel, LD led to degradation of a DELLA protein, SLENDER (within 48 h of LD or within 2 h of GA application). There was no effect on GA catabolism or abscisic acid content. Loss of SLENDER, which is a repressor of GA signaling, confirms the physiological relevance of increased GA content in LD. For flowering, applied GA replaced the need for LD but not that for vernalization. Thus, GAs may be an LD, leaf-sourced hormonal signal for flowering of L. perenne. By contrast, vernalization had little impact on GA or SLENDER levels or on SLENDER degradation following GA application. Thus, although vernalization and GA are both required for flowering of L. perenne, GA signaling is independent of vernalization that apparently impacts on unrelated processes.     

NADP-Dependent Phenylacetaldehyde Dehydrogenase for Degradation of Phenylethylamine in Arthrobacter globiformis

The role of endogenous gibberellin (GA) in the flowering of the short-day plant, Pharbitis nil, was investigated by using uniconazole, which is a specific inhibitor of GA biosynthesis. Both the endogenous GA level and flowering response decreased with increasing concentration of uniconazole applied via the roots. The strongest inhibition of flowering was observed when uniconazole was applied one day before a 15-h dark treatment. The inhibition by uniconazole was overcome by an application of GAs to the plumules, the order of effectiveness of the endogenous GAs in P. nil being GA₁ ≧GA₂₀>GA₁₉≧GA₄₄>GA₅₃»GA_H. This is the first report of the correlation between the endogenous GA level and flowering response in P. nil. It was found that endogenous GAs were required for the flowering of P. nil during or just after the dark period.     

Timing of Photoperiodic Flowering:Light Perception and Circadian Clock

Flowering symbolizes the transition of a plant from vegetative phase to reproductive phase and is controlled by fairly complex and highly coordinated regulatory pathways. Over the last decade, genetic studies in Arabidopsis have aided the discovery of many signaling components involved in these pathways. In this review, we discuss how the timing of flowering is regulated by photoperiod and the involvement of light perception and the circadian clock in this process. The specific regulatory mechanisms on CONSTANS expression and CONSTANS stability by the circadian clock and photoreceptors are described in detail. In addition, the roles of CONSTANS, FLOWERING LOCUS T, and several other light signaling and circadiandependent components in photoperiodic flowering are also highlighted.     

Timing of Photoperiodic Flowering：Light Perception and Circadian Clock

Flowering symbolizes the transition of s plant from vegetative phase to reproductive phase and is controlled by fairly complex and highly coordinated regulatory pathways. Over the last decade, genetic studies in Arabidopsis have aided the discovery of many signaling components involved in these pathways. In this review, we discuss how the timing of flowering is regulated by photoperiod and the involvement of light perception and the circadian clock in this process. The specific regulatory mechanisms on CONSTANS expression and CONSTANS stability by the circadian clock and photoreceptors are described in detail. In addition, the roles of CONSTANS, FLOWERING LOCUS T, and several other light signaling and circadian-dependent components in photoperiodic flowering are also highlighted.     

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.     

水稻开花期调控的分子机理研究进展

水稻准确地感知外部环境信号,通过内部复杂的基因网络做出反应,在一年中最适合的时候开花繁殖。与长日促进长日模式植物拟南芥开花相反,短日促进短日模式植物水稻开花。通过对水稻和拟南芥的开花期调控机理的对比分析,发现水稻和拟南芥有着一些相对保守的开花期控制基因,其调控机理也是相似的。另外,水稻也有一些独特的开花期控制基因和开花途径。本文着重从光周期对水稻开花期的调控途径和作用机理角度进行了阐述,并对水稻开花期的自然变异与其育种应用、生物钟关联基因、光中断现象和临界日长现象以及开花期与产量的关系进行了总结。