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The transcription factor FUSCA3 controls developmental timing in Arabidopsis through the hormones gibberellin and abscisic acid.

Although plants continually produce different organs throughout their life cycle, little is known about the factors that regulate the timing of a given developmental program. Here we report that the restricted expression of FUS3 to the epidermis is sufficient to control foliar organ identity in Arabidopsis by regulating the synthesis of two hormones, abscisic acid and gibberellin. These hormones in turn regulate the rates of cell cycling during organ formation to determine whether an embryonic or adult leaf will emerge. We also show that FUS3 expression is influenced by the patterning hormone, auxin, and therefore acts as a nexus of hormone action during embryogenesis. The identification of lipophillic hormones downstream of a heterochronic regulator in Arabidopsis has parallels to mechanisms of developmental timing in animals and suggests a common logic for temporal control of developmental programs between these two kingdoms.

Abscisic Acid↗

A DELLAcate balance: the role of gibberellin in plant morphogenesis.

The importance of gibberellin (GA) in vegetative and reproductive development has been known for some time. Recent studies have uncovered new roles of GA in leaf differentiation, photomorphogenesis and pollen-tube growth. Significant contributions to our understanding of GA-regulated morphogenesis include the identification of upstream regulators of GA biosynthesis, the elucidation of the function of GA signaling components, and the isolation of downstream targets. In addition, the mechanisms of interactions between GA and other hormone pathways are beginning to be revealed at the molecular level.

Gene Expression Regulation, Developmental↗

Turning on gibberellin and abscisic acid signaling.

The phytohormones gibberellin (GA) and abscisic acid (ABA) play essential and often antagonistic roles in regulating plant growth, development and stress responses. The long-awaited identification of receptors for both GA and ABA has shed light upon the initial events that surround the perception of these two phytohormones. The discovery of these receptors also challenges conventional views of plant hormone signaling and raises intriguing questions regarding the nature of GA and ABA perception and the initiation of their signaling pathways. Moreover, recent advances in understanding GA and ABA signaling point to the existence of multiple, non-linear cell- and compartment-specific pathways that regulate genomic and non-genomic responses to these phytohormones.

Abscisic Acid↗

Kaurenolides and fujenoic acids are side products of the gibberellin P450-1 monooxygenase in Gibberella fujikuroi.

The steps involved in kaurenolide and fujenoic acids biosynthesis, from ent-kauradienoic acid and ent-6alpha,7alpha-dihydroxykaurenoic acid, respectively, are demonstrated in the gibberellin (GA)-deficient Gibberella fujikuroi mutant SG139, which lacks the entire GA-biosynthesis gene cluster, complemented with the P450-1 gene of GA biosynthesis (SG139-P450-1). ent-[2H]Kauradienoic acid was efficiently converted into 7beta-hydroxy[2H]kaurenolide and 7beta,18-dihydroxy[2H]kaurenolide by the cultures while 7beta-hydroxy[2H]kaurenolide was transformed into 7beta,18-dihydroxy[2H]kaurenolide. The limiting step was found to be hydroxylation at C-18. In addition, SG139-P450-1 transformed ent-6alpha,7alpha-dihydroxy[14C4]kaurenoic acid into [14C4]fujenoic acid and [14C4]fujenoic triacid. Fujenal was also converted into the same products but was demonstrated not to be an intermediate in this sequence. All the above reactions were absent in the mutant SG139 and were suppressed in the wild-type strain ACC917 by disruption of the P450-1 gene. Kaurenolide and fujenoic acids synthesis were associated with the microsomal fraction and showed an absolute requirement for NADPH or NADH, all properties of cytochrome P450 monooxygenases. Only 7beta-hydroxy[14C4]kaurenolide synthesis and not further 18-hydroxylation was detected in the microsomal fraction. The substrates for the P450-1 monooxygenase, ent-kaurenoic acid and [2H]GA12, efficiently inhibited kaurenolide synthesis with I50 values of 3 and 6 microM, respectively. Both substrates also inhibited ent-6alpha,7alpha-dihydroxy[14C4]kaurenoic acid metabolism by SG139-P450-1. Conversely, [14C4]GA14 synthesis from [14C4]GA12-aldehyde was inhibited by ent-[2H]kauradienoic acid and fujenal with I50 values of 10 and 30 microM, respectively. These results demonstrate that kaurenolides and seco-ring B kaurenoids are formed by the P450-1 monooxygenase (GA14 synthase) of G. fujikuroi and are thus side products that probably result from stabilization of radical intermediates involved in GA14 synthesis.

Aldehydes↗

Effects of ABA, auxin, and gibberellin on the expression of genes for vacuolar H+ -inorganic pyrophosphatase, H+ -ATPase subunit A, and Na+/H+ antiporter in barley.

We analyzed the effects of ABA, auxin, and gibberellin on the expression of two genes (HVP1 and HVP10) for vacuolar H(+)-inorganic pyrophosphatase (EC 3.6.1.1) and one (HvVHA-A) for the catalytic subunit (subunit A) of vacuolar H(+)-ATPase (EC 3.6.1.3) by quantification of the transcript levels, to identify the hormones responsible for regulating the expression of these genes in barley (Hordeum vulgare L.) in response to environmental changes such as salt stress. ABA markedly induced the expression of HVP1 and slightly increased the expression of HVP10 and HvVHA-A. In contrast, 2,4-D only increased the expression of HVP1, and GA(3) had no significant effects on any gene. The maximum level of HVP1 transcripts in response to these hormones was also much higher than the levels of HVP10 and HvVHA-A transcripts. In addition, we also analyzed the expression of one gene (HvNHX1) for vacuolar Na(+)/H(+) antiporter, and HvNHX1 expression changed in a pattern similar to that of the HVP1 expression. Furthermore, treatment with ABA and 2,4-D increased Na(+)/H(+) antiport activity and proton-translocating activities by H(+)-PPase and H(+)-ATPase in tonoplast vesicles, and treatment with ABA also increased the amount of V-PPase protein of tonoplast vesicles. These results suggest that the hormones ABA and 2,4-D regulate the expression of the H(+)-pump and Na(+)/H(+) antiporter genes and are thus important effectors that regulate the expression of HVP1and HvNHX1.

2,4-Dichlorophenoxyacetic Acid↗

A role for the ubiquitin-26S-proteasome pathway in gibberellin signaling.

The gibberellin (GA) signaling pathway, like auxin and jasmonate signaling, uses the ubiquitin-proteasome pathway to control expression through protein degradation. A conserved F-box protein of an SCF E3 ubiquitin ligase is a positive regulator of GA signaling in Arabidopsis and rice. GA apparently stimulates stem elongation by causing this SCF complex to regulate negatively a family of negative regulators of GA response (the DELLA family of putative transcription factors). The DELLA family members AtRGA or (Repressor of ga1-3) and OsSLR1 (SLENDER RICE1) proteins both appear to be subject to GA-induced proteolysis. The need to have the F-box genes AtSLY1 and OsGID2 for this proteolysis suggests that GA causes proteolysis of AtRGA/OsSLR1 via the SCF(AtSLY1/OsGID2) ubiquitin ligase.

Amino Acid Sequence↗

Tall tales from sly dwarves: novel functions of gibberellins in plant development.

Gibberellins (GAs) are endogenous hormones controlling numerous aspects of plant growth and development. Our present understanding of GA physiology is based largely on genetic analysis in model plants such as Arabidopsis. In spite of the success of this approach, the discovery of additional physiological roles for GAs in seed development, pollen tube growth and meristem development indicates that the existing collection of GA-related mutants (identified partially or entirely on the basis of vegetative phenotypes) has failed to uncover all aspects of plant development that are controlled by GAs. The continued use of ever improving forward and reverse genetic techniques is expected to lead to the discovery of further novel roles for GAs in plant development.

Arabidopsis↗

Gibberellin biosynthesis: metabolic evidence for three steps in the early 13-hydroxylation pathway of rice.

[14C4]GA53, [14C4]GA44, and [2H2/14C4]GA19 were injected separately into seedlings of rice (Oryza sativa) using a dwarf mutant (d35) that has low levels of endogenous gibberellins (GAs). After 8 h incubation, the shoots were extracted and the labeled metabolites were identified by full-scan gas chromatography mass spectrometry (GC-MS) and Kovats retention indices (KRIs). Our results document the metabolic sequence, GA53-->GA44-->GA19-->GA20 and the presence of endogenous GA53, GA44, GA19, GA20 and GA1. Previous metabolic studies have shown the presence of the step, GA20-->GA1 in rice. Taken together, the data establish in vegetative shoots of rice the presence of the early 13-hydroxylation pathway, a pathway that originates from GA12 and leads to bioactive GA1.

Gas Chromatography-Mass Spectrometry↗

Gibberellins in shoots and developing capsules of Populus species.

Extracts of stems of growing shoots of Populus deltoides and P. trichocarpa, and developing capsules of P. deltoides were analysed for gibberellins (GAs) by gas chromatography-mass spectrometry. The following known GAs were identified by comparison of their Kovats retention indices (KRIs) and mass spectra with those of standards: GA1, GA8, GA9, GA19, GA20, 16 beta,17-dihydro-17-hydroxy GA20, GA23, GA28, GA29, GA34, GA44, and GA97. Several of these have not been previously reported from Populus. In addition, two new GAs were identified as 12 beta-hydroxy GA53 (GA127) and 16 beta,17-dihydro-17-hydroxy GA53 and their structures were confirmed by partial synthesis. Evidence was found of 16,17-dihydro-16,17-dihydroxy GA9, 16,17-dihydro-16,17-dihydroxy GA12, 12-hydroxy GA14, and GA34-catabolite by comparison of mass spectra and KRIs with published data. Several putative GAs (hydroxy- and dihydroxy-GA12-like) were also found. The catabolites of active GAs or of key precursors, hydroxylated at C-2 in stems and either C-2, C-12, C-17, or C-16,17 in capsules, were the major proportion of the GAs.

Gas Chromatography-Mass Spectrometry↗

3 beta,13-dihydroxylated C20 gibberellins from inflorescences of Rumex acetosa L.

Using full scan GC-MS a wide range of gibberellins (GAs) was identified in the young inflorescences of the dioecious species Rumex acetosa L., consistent with the ubiquitous early 13-hydroxylation pathway in both male and female plants. In addition, R. acetosa is the first species in which all three 3beta,13-dihydroxylated C(20)-GAs-GA(18), GA(38) and GA(23)-have been identified in the same organism, suggesting an early 3beta,13-dihydroxylation biosynthesis pathway in this species. Authentic GA(18), GA(38) and GA(23) were synthesized and their effects and that of GA(1), a GA common to both pathways, on the time to inflorescence emergence was investigated. GA(1) accelerated the emergence of inflorescences in both male and female plants. In addition some evidence for biological activity per se of the C(20)-GA(38) was obtained.

Flowers↗

Synthesis and confirmation of structure for a new gibberellin, 2 beta-hydroxy-GA12 (GA110), from spinach and oil palm.

The identity of a new gibberellin (GA) in spinach and oil palm sap has been confirmed as 2 beta-hydroxy-GA12 (GA110) by comparisons of GC-mass spectral data obtained for the trimethylsilyl ether methyl ester derivatives with those of a synthetic sample prepared by means of a 24 step sequence from gibberellic acid; 2 beta-hydroxy-GA24 was also prepared. Experimental details for the latter part of the syntheses are described.

Gas Chromatography-Mass Spectrometry↗

Gibberellins in seedlings and flowering trees of Prunus avium L.

Extracts of acids from mature seeds, germinating seeds, first, second and third year seedlings as well as mature, flowering trees of sweet cherry (Prunus avium L. cv. Stella) were analysed by gas chromatography-mass spectrometry. The presence of the known gibberellins (GAs) GA1 (1), GA3 (4), GA5 (7), GA8 (11), GA19 (14), GA20 (12), GA29 (13), GA32 (5), GA85 (2), GA86 (3) and GA87 (6) was confirmed by comparison of their mass spectra and Kovats retention indices with those of standards or literature values. In addition, 16alpha,17-dihydrodihydroxy GA25 (16) was identified and its stereochemistry confirmed by rational synthesis. The 12alpha,13-dihydroxy GAs, GA32 (5), GA86 (2), GA86 (3) and GA87 (6), were detected in mature seeds, germinating seeds and young seedlings, but not in flowering plants. The 13-hydroxy GAs, GA1 (1) and GA3 (4), were present in germinating seeds and, in addition to these, GA5 (7), GA8 (11), GA19 (14), GA20 (12) and GA29 (13) were detected in seedlings and mature flowering plants. In germinating seeds and seedlings (while the plants were growing actively), concentrations of the 12alpha,13-dihydroxy GAs, measured by bioassay, declined and those of the 13-hydroxy GAs increased. The results are discussed with reference to the known and predicted effects of the GAs on the vegetative growth and flowering of P. avium plants.

Chromatography, High Pressure Liquid↗

Gibberellin metabolism and signaling.

Gibberellins (GAs) are a family of plant hormones controlling many aspects of plant growth and development including stem elongation, germination, and the transition from vegetative growth to flowering. Cloning of the genes encoding GA biosynthetic and inactivating enzymes has led to numerous insights into the developmental regulation of GA hormone accumulation that is subject to both positive and negative feedback regulation. Genetic and biochemical analysis of GA-signaling genes has revealed that posttranslational regulation of DELLA protein accumulation is a key control point in GA response. The highly conserved DELLA proteins are a family of negative regulators of GA signaling that appear subject to GA-stimulated degradation through the ubiquitin-26S proteasome pathway. This review discusses the regulation of GA hormone accumulation and signaling in the context of its role in plant growth and development.

Gibberellins↗

Identification of Dof proteins with implication in the gibberellin-regulated expression of a peptidase gene following the germination of rice grains.

Type III carboxypeptidase (CPD3) is one of the hydrolytic enzymes whose expression is up-regulated by gibberellins (GA) in the aleurones of germinated cereal grains. A number of pyrimidine boxes and a sequence resembling the gibberellic acid response element (GARE) are observed in the region upstream of the transcription initiation site of the CPD3 gene, showing a characteristic of cereal GA-responsive genes. Transient gene expression assays in germinated rice aleurone demonstrated that the CPD3 promoter was able to confer hormonally responses on the expression of the reporter gene. By southwestern screening, several cDNAs encoding the Dof class proteins were isolated from a rice aleurone library. Each mRNA accumulation for five novel members of Dof proteins (OsDof1--5) occurs with a different time course and in a tissue-specific manner following the germination of grains. Of these, the expression of the OsDof3 gene is abundant in aleurones where it precedes that of the CPD3 gene, implying that this is an early response gene of GA. The OsDof3 protein, expressed in Escherichia coli, selectively bound AAAG motifs of the pyrimidine boxes through the DNA-binding activity of its Dof domain. Co-expression experiments in aleurones suggested that the OsDof3 protein should play a regulatory role in the expression of the CPD3 gene under the control of GA.

Amino Acid Sequence↗

The gibberellin pathway mediates KNOTTED1-type homeobox function in plants with different body plans.

BACKGROUND: The shoot apical meristem (SAM) is an indeterminate structure that gives rise to the aerial parts of higher plants. Leaves arise from the differentiation of cells at the flanks of the SAM. Current evidence suggests that the precise regulation of KNOTTED1-like homeobox (KNOX) transcription factors is central to the acquisition of leaf versus meristem identity in a wide spectrum of plant species. Factors required to repress KNOX gene expression in leaves have recently been identified. Additional factors such as the CHD3 chromatin remodeling factor PICKLE (PKL) act to restrict meristematic activity in Arabidopsis leaves without repressing KNOX gene expression. Less is known regarding downstream targets of KNOX function. Recent evidence, however, has suggested that growth regulators may mediate KNOX activity in a variety of plant species. RESULTS: Here we show that reduced activity of the gibberellin (GA) growth regulator pathway promotes meristematic activity, both in the natural context of KNOX function in the SAM and upon ectopic KNOX expression in Arabidopsis leaves. We show that constitutive signaling through the GA pathway is detrimental to meristem maintenance. Furthermore, we provide evidence that one of the functions of the KNOX protein SHOOTMERISTEMLESS (STM) is to exclude transcription of the GA-biosynthesis gene AtGA20ox1 from the SAM. We also demonstrate that AtGA20ox1 transcript is reduced in the pkl mutant in a KNOX-independent manner. Moreover, we show a similar interaction between KNOX proteins and GA-biosynthesis gene expression in the tomato leaf and implicate this interaction in regulation of the dissected leaf form. CONCLUSIONS: We suggest that repression of GA activity by KNOX transcription factors is a key component of meristem function. Transfer of the KNOX/GA regulatory module from the meristem to the leaf may have contributed to the generation of the diverse leaf morphologies observed in higher plants.

Arabidopsis↗

Gibberellin signal transduction.

Recent studies using biochemical and genetic approaches have identified a number of components, including several negative regulators, of the gibberellin (GA) signal transduction pathway in higher plants. The basal state of GA signaling is likely to be repressive, and the GA signal seems to activate the pathway by de-repression to allow GA-stimulated growth and development.

Arabidopsis↗

Gibberellin signalling pathway.

Recent molecular biological and genetical studies have identified several positive and negative regulators of gibberellin (GA) signalling pathways in higher plants. The DELLA protein functions as a negative regulator of GA signalling; its degradation through the ubiquitin/proteasome pathway is a key event in the regulation of GA-stimulated processes.

Gene Expression Regulation, Developmental↗

Regulation of gibberellin biosynthesis by light.

Phytochromes regulate transcript levels of gibberellin biosynthesis enzymes, GA 20-oxidases and/or GA 3beta-hydroxylases, in germinating lettuce and Arabidopsis seeds and in de-etiolating pea seedlings. Feedback regulation of GA biosynthesis by active GA is well established, but other mechanisms for regulation of these biosynthetic genes also exist, as this feedback does not operate on a GA 3beta-hydroxylase gene of Arabidopsis during seed germination.

Arabidopsis↗