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The effect of carbon and nitrogen sources on auxins and gibberellin-like substances synthesis by bacteria isolated from the roots of pine seedlings (Pinus silvestris L).

It was found that in the presence of sodium succinate synthesis of auxins and gibberellin-like substances by microflora in the rhizosphere of pine seedlings was higher than in the presence of the other carbon sources. The smallest amounts of these substances were found in media with sodium pyruvate. The highest C-content (C:N = 60:1) was not optimal for the synthesis of plant growth regulators. Ammonia ions inhibited the production of gibberellin-like substances. The effect of different nitrogen sources and C:N ratio on the synthesis of auxins was different depending on bacterial strain used.

Ammonia↗

Production of gibberellin-like substances by bacteria and fungi isolated from the roots of pine seedlings (Pinus silvestris L.).

The production of gibberellin-like substances by bacteria and fungi was investigated. A considerable number of the microorganisms studied produced these substances (55 and 86% of bacterial and fungal isolates respectively). Most of the organisms tested synthesized biologically active substances located on the chromatograms with the solvent system benzene, acetic acid (10:3 v/v) at Rf 0.05--0.3. The ammount of gibberellin-like substances produced varied in the range 0.00007--0.00065 GA3 equivalents mug/ml in bacteria and 0.00014--0.00118 mug/ml in fungi.

Actinomycetales↗

Inhibition of gibberellin-induced elongation growth of rice by feruloyl oligosaccharides.

The biological activity of cell wall-derived feruloyl oligosaccharides was investigated using a modified micro-drop bioassay. A feruloyl arabinoxylan trisaccharide (FAXX) and a feruloyl arabinoxylan tetrasaccharide (FAXXX) were found to inhibit the gibberellin-induced elongation of dwarf rice (Oryza sativa L., cv, Tan-ginbozu) that had been treated with uniconazole (S-3307), an inhibitor of the biosynthesis of gibberellins. An arabinoxylan trisaccharide (AXX) was ineffective. The growth-inhibitory effect of feruloyl oligosaccharides depended on their feruloyl and glycosyl moieties. The amount of esterified diferulic acid residues in the cell walls of the second leaf sheath of rice seedlings that had been treated with FAXX was almost same as that of controls. Feruloyl oligosaccharides did not inhibit the incorporation of [14C]leucine into acid-precipitable proteins by suspension-cultured maize cells, whereas cinnamic acid and its derivatives strongly inhibited such incorporation.

Carbohydrate Sequence↗

Conditioning of Fasciation by Gibberellin and Genotype in Cotton (Gossypium hirsutum L.).

The round-leafed mutant cotton line L-501 developed fasciation of the upper stem when field grown in Central Asia. Fasciation co-segregated with the mutant gene for round leaves In.(l) Fasciation developed at the flowering stage, but removal of floral buds did not prevent fasciation. Fasciation in L-501 could be inhibited by the gibberellin (GA) biosynthesis inhibitor chlorocholine chloride or by fusicoccin. GA(3) application in the field induced fasciation in the mutant's parental line L-463, which has five-lobed leaves and does not normally develop fasciation. Fasciation did not develop in either line, even after GA(3) treatment, in UK glasshouse conditions.Key Words. Fasciation-Gibberellin-Gossypium-Leaf-shape-Mutanthttp://link.springer-ny.com/link/service/journals/00344/bibs/18n1p45.html

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Gibberellins and Subapical Cell Divisions in Relation to Bud Set and Bud Break in Salix pentandra.

In young plants of Salix pentandra, a temperate zone deciduous woody species, elongation growth ceases and a terminal bud is formed at day lengths shorter than a critical length. This is the first step in dormancy development, making survival under harsh winter conditions possible. Early studies strongly indicate that gibberellin is involved in the photoperiodic control of bud set and bud break. GA(1) action was studied by application under short days to plants where cessation of shoot elongation had occurred, followed by subsequent anatomic investigations of shoot tips. Under short days the frequency of cell division decreased rapidly along with the earlier observed decrease in GA(1) levels. Application of GA(1) to short-day-induced terminal buds rapidly stimulated cell division in apices several days before visible shoot elongation in response to this treatment was observed. One day after GA(1) application a fourfold increase in cell division frequency in apices was observed, increasing to a maximum of sevenfold 2 days after application. Long-day treatment leading to induction of bud break after about 4-6 days was followed by slowly increasing frequency of cell divisions. In earlier studies of this species, short days and gibberellins had no effect on cell elongation. These data show that increased GA(1) content, by application or long-day treatment, results in increased frequency of mitosis. This strongly indicates that GA(1) affects stem elongation in connection with bud set and bud break primarily by affecting cell divisions in subapical tissues.

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Remarkable control of radical cyclization processes of cyclic enyne: total syntheses of (+/-)-methyl gummiferolate, (+/-)-methyl 7beta-hydroxykaurenoate, and (+/-)-methyl 7-oxokaurenoate and formal synthesis of (+/-)-gibberellin a(12) from a common synthetic precursor.

Total syntheses of (+/-)-methyl gummiferolate (13b), (+/-)-methyl 7beta-hydroxykaurenoate (14b), and (+/-)-methyl 7-oxokaurenoate (14d) and a formal synthesis of (+/-)-gibberellin A(12) (15) have been accomplished through the common synthetic precursor, (3aR,7aR)-3,3-dimethyl-7a-(2-propynyl)-3a,4,7,7a-tetrahydroisobenzofuranone (16). The homoallyl-homoallyl radical rearrangement reaction of the monocyclic enyne 25, derived from 16 in two steps, afforded the bicyclo[2.2.2]octane compound 26, which was converted to (+/-)-methyl gummiferolate (13b). In contrast, the radical cyclization of the bicyclic enyne 16 gave the tricyclic lactone 19, leading to (+/-)-methyl 7beta-hydroxykaurenoate (14b) and (+/-)-methyl 7-oxokaurenoate (14d). Transformation of 14d into lactone 20 was carried out in a single step under bromination conditions. This constitutes a formal total synthesis of gibberellin A(12) (15).

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Gibberellins in relation to flowering in Polianthes tuberosa.

Endogenous gibberellins (GAs) in corms of Polianthes tuberosa L. (cv. Double) were isolated and identified by high performance liquid chromatography, bioassay and combined capillary gas chromatography-mass spectrometry (GC-MS). Gibberellins A1, A19, A20 and A53 were quantified at the vegetative, early floral initiation and flower development stages. The identification of 13-hydroxylated GAs indicates the presence of the early 13-hydroxylation pathway in P. tuberosa corms. An increase in GA1 and GA20, and a decrease in GA19 levels, coincided with the transition from the vegetative phase to the stages of early floral initiation and flower development. GA53 stayed at constant levels at the 3 different growth stages. The absence of GA1 in vegetative corms and its presence in corms at early floral initiation and flower development stages suggest that GA1 is a causal factor in inducing floral initiation in P. tuberosa. When GA1, GA3, GA4, GA20 and GA32 were applied to corms at the vegetative stage (plants about 5 cm in height), floral initiation was promoted by all of the GAs used, GA32 being the most active. In contrast with the other GAs, GA32 had no effect on stem elongation. Therefore, it is suggested that hydroxylated C-19 GAs play an important role in flower induction in P. tuberosa.

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Hormonal regulation of a cysteine proteinase gene, EPB-1, in barley aleurone layers: cis- and trans-acting elements involved in the co-ordinated gene expression regulated by gibberellins and abscisic acid.

The synthesis of EPB, a cysteine proteinase responsible for the degradation of seed endosperm storage proteins in barley (Hordeum vulgare), is induced by gibberellins (GA) and repressed by abscisic acid (ABA). The EPB gene family consists of two very similar members, EPB-1 and EPB-2, with the former being more highly induced by GA. We have functionally characterized the cis-acting elements in the EPB-1 promoter and determined that a gibberellin response element (GARE), a pyrimidine box and an upstream element are necessary for GA induction. By comparison with the promoters of alpha-amylase genes, which are also induced by GA, we suggest that GARE is coupled with the upstream element and the pyrimidine box to form a GA response complex. In addition, we have shown that the 3'-untranslated/untranscribed region of the EPB-1 gene is required for a low background expression in the absence of GA. Constitutive expression of a transcription factor, GAMyb, in the absence of GA leads to the transactivation of EPB-1 expression in a dosage dependent manner with the highest level comparable to that in fully GA-induced tissue. Co-expression of a truncated version of GAMyb containing only the DNA binding domain blocks the GA-induction of EPB-1, further supporting the role of GAMyb in the regulation of gene expression. Although ABA is very effective in blocking the GA induction of EPB-1, it has no effect on the GAMyb-mediated expression of EPB-1. We suggest that ABA acts upstream of the formation of functional GAMyb which co-ordinates the hormonal regulation of a diverse group of genes in cereal aleurone layers, including those encoding EPB and alpha-amylases.

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Gibberellin 2-oxidation and the SLN gene of Pisum sativum.

Two cDNAs encoding gibberellin 2-oxidases were isolated from maturing pea seeds. The first, PsGA2ox1, was isolated by activity screening of a Lambda-ZAP cDNA library excised into phagemid form and expressed in Escherichia coli. The second, PsGA2ox2, was obtained initially as a PCR product using degenerate primers designed according to conserved regions of plant 2-oxoglutarate-dependent dioxygenases. E. coli heterologous expression products of PsGA2ox1 and PsGA2ox2 converted GA1 to GA8, as shown by HPLC-radiocounting, and gas chromatography-MS. PsGA2ox1 converted GA20 to GA29, but GA20 was a poor substrate for the PsGA2ox2 expression product. Furthermore, PsGA2ox1 converted GA29 to GA29-catabolite at a low level of efficiency while PsGA2ox2 did not catalyse this step. A cDNA of PsGA2ox1 isolated from plants of genotype sln contained a single base deletion which was predicted to produce a truncated protein and gibberellin 2-oxidase activity could not be demonstrated from this cDNA. A 10 bp size difference between the introns of the SLN and sln PsGA2ox1 genes was used to show co-segregation between the SLN and sln phenotypes and the size of the PCR products. PsGA2ox1 transcripts were more abundant in cotyledons than in shoots, while the reverse was the case for PsGA2ox2. The expression patterns of the genes, together with the effects of the sln mutation, indicate that PsGA2ox1 plays a major role in GA20 deactivation in both shoots and maturing seeds, while the PsGA2ox2 gene might be important for GA1 deactivation in the shoot.

Amino Acid Sequence↗

The dwarf-1 (dt) Mutant of Zea mays blocks three steps in the gibberellin-biosynthetic pathway.

In plants, gibberellin (GA)-responding mutants have been used as tools to identify the genes that control specific steps in the GA-biosynthetic pathway. They have also been used to determine which native GAs are active per se, i.e., further metabolism is not necessary for bioactivity. We present metabolic evidence that the D1 gene of maize (Zea mays L.) controls the three biosynthetic steps: GA20 to GA1, Ga20 to GA5, and GA5 to GA3. We also present evidence that three gibberellins, GA1, GA5, and GA3, have per se activity in stimulating shoot elongation in maize. The metabolic evidence comes from the injection of [17-13C,3H]GA20 and [17-13C,3H]GA5 into seedlings of d1 and controls (normal and d5), followed by isolation and identification of the 13C-labeled metabolites by full-scan GC-MS and Kovats retention index. For the controls, GA20 was metabolized to GA1,GA3, and GA5; GA5 was metabolized to GA3. For the d1 mutant, GA20 was not metabolized to GA1, GA3, or to GA5, and GA5 was not metabolized to GA3. The bioassay evidence is based on dosage response curves using d1 seedlings for assay. GA1, GA3, and GA5 had similar bioactivities, and they were 10-times more active than GA20.

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Rice gibberellin-insensitive dwarf mutant gene Dwarf 1 encodes the alpha-subunit of GTP-binding protein.

A rice Dwarf 1 gene was identified by using a map-based cloning strategy. Its recessive mutant allele confers a dwarf phenotype. Linkage analysis revealed that a cDNA encoding the alpha-subunit of GTP-binding protein cosegregated with d1 in 3,185 d1 segregants. Southern hybridization analysis with this cDNA as a probe showed different band patterns in several d1 mutant lines. In at least four independent d1 mutants, no gene transcript was observed by Northern hybridization analysis. Sequencing analysis revealed that an 833-bp deletion had occurred in one of the mutant alleles, which resulted in an inability to express GTP-binding protein. A transgenic d1 mutant with GTP-binding protein gene restored the normal phenotype. We conclude that the rice Dwarf 1 gene encodes GTP-binding protein and that the protein plays an important role in plant growth and development. Because the d1 mutant is classified as gibberellin-insensitive, we suggest that the GTP-binding protein might be associated with gibberellin signal transduction.

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Gibberellin A(4/7) promotes cone production on potted grafts of eastern white pine.

Potted grafts of eastern white pine (Pinus strobus L.) growing in a heated greenhouse were sprayed weekly with 0, 250 or 500 mg l(-1) of gibberellin A(4/7) for four different 6-week periods between mid-May and mid-September. At different periods, gibberellin A(4/7) concentrations of 250 and 500 mg l(-1) were effective in promoting pollen- and seed-cone production. Spraying in May and June, during the period of rapid terminal shoot elongation, significantly increased pollen-cone production, whereas spraying in August and September, about a month after terminal shoot elongation, significantly increased seed-cone production.

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Metabolism of deuterium- and tritium-labeled gibberellins in cambial region tissues of Eucalyptus globulus stems.

Deuterium- and tritium-labeled gibberellins (GAs) were applied to stems of 3-year-old Eucalyptus globulus Labill. saplings and 9-month-old potted seedlings. Cambial region tissues surrounding the application point were collected 6, 24 or 48 h later. Twenty-four hours after application of 5 &mgr;g of [(2)H(2)]GA(20), 7% of the cambial region GA(20) pool, 7% of the GA(1) pool and 58% of the GA(29) pool were labeled with deuterium based on selected ion monitoring of purified extracts subjected to gas chromatography-mass spectrometry. The relatively low percent dilution of endogenous GAs by [(2)H(2)]GAs suggests that the exogenous application of [(2)H(2)]GA(20) did not result in substrate overloading, indicating that these conversions probably occur naturally within cambial region tissues. Extracts from similar cambial region tissues fed tritium-labeled GAs were sequentially fractionated by SiO(2) partition chromatography, C(18) reversed phase HPLC and N(CH(3))(2) HPLC. The radioactivity profiles indicated metabolism of GA(20) to GA(1) and GA(29), GA(1) conversion to GA(8), GA(4) to GA(34) and GA(9) to GA(51). Gibberellins GA(34), GA(51) and GA(29) are C-2beta-hydroxylated catabolites of low biological activity, whereas GA(1) and GA(4) are probably effectors of growth in the Eucalyptus stem and shoot. Evidence for C-13 hydroxylation of GA(4) to GA(1), GA(9) to GA(4) or GA(9) to GA(20) in the stem was inconclusive. Thus, although GA(4) and GA(9) are native to cambial region tissues, GA(1) is probably not produced from them in significant quantities. We conclude that the early C-13-hydroxylation pathway; i.e., conversion of GA(19) to GA(20) to GA(1), is the major pathway of GA(1) biosynthesis.

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Influence of water table decline on growth allocation and endogenous gibberellins in black cottonwood.

Cottonwoods occur in riparian areas where water table depth generally varies with the elevation of the adjacent river. Plant adaptation to the riparian zone requires the coordination of root elongation to maintain contact with the water table during the summer decline. We investigated the effects of rate of water decline on growth allocation and concentrations of endogenous gibberellins (GAs) in black cottonwood (Populus trichocarpa Torr. & A. Gray ex Hook.) saplings. Rhizopods were used to achieve water decline rates of 0, 2 and 4 cm day(-1). Root elongation approximately doubled in response to the 2 cm day(-1) treatment, whereas leaf area was reduced. A water decline rate of 4 cm day(-1) led to water stress, as evidenced by reduced growth, increased leaf diffusive resistance, decreased water potential, and leaf senescence and abscission. Endogenous GAs were extracted, purified and analyzed by gas chromatography-selected ion monitoring with internal [(2)H(2)]GA standards. Across the sampled plant organs, GAs were generally highest in shoot tips and sequentially lower in basal stems, root tips, leaves and upper roots; GAs were thus abundant in rapidly growing tissues. Of the GAs measured, GA(1) tended to predominate, followed sequentially by GA(3), GA(8), GA(19), GA(20), GA(29) and GA(4). There was little relationship between GA concentration and growth allocation across the water table decline treatments, although GA(8) was consistently reduced in plants experiencing water table decline. Because GA(8) is the final gibberellin in the metabolic sequence, it might be useful for assessing historic patterns of GAs and growth rate. This study demonstrated changes in growth allocation in response to water table decline, but provided little evidence that endogenous GAs play a primary role in the regulation of root elongation in response to water table decline.

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Inhibition of Gibberellin Production in the Fungi Gibberella fujikuroi and Sphaceloma manihoticola by Plant Growth Retardants.

The effect of different types of plant growth retardants on fungal gibberellin (GA) formation has been studied in cultures of Gibberella fujikuroi and Sphaceloma manihoticola. Quaternary ammonium compounds (chlormequat chloride, mepiquat chloride, Amo-1618), triazoles (uniconazole and several experimental compounds), and the norbornanodiazetine tetcyclacis inhibited GA biosynthesis in both fungal species. Concentrations between 2 x 10(-4) and 10(-9)m were required for a 50% inhibition of the production of gibberellin A(3) in Gibberella fujikuroi and of giberellin A(4) in Sphaceloma manihoticola. The formation of other prominent GAs was affected at a similar degree of intensity. Tetcyclacis was the most active compound in both fungi. Compared to the growth retardants mentioned above, the biological activity of chlorphonium chloride was low. The acylcyclohexanediones prohexadione and LAB 198 999 had virtually no activity. Most likely, this lack of activity is due to a rapid metabolism of the compounds in the cultures. For the triazole-type compounds and tetcyclacis, a relatively distinct correlation exists in their ability to inhibit GA formation in fungal cultures, to block ent-kaurene oxygenase in a cell-free system, and to reduce shoot growth of rice seedlings. Due to differences in their metabolic fate and species specificities, such conclusions cannot be made for the other compounds.

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Use of an Acylcyclohexanedione Growth Retardant, LAB 198 999, to Determine Whether Gibberellin A(20) Has Biological Activity per se in Dark-Grown Dwarf (le) Seedlings of Pisum sativum.

Dwarf (le(5839)) seedlings of Pisum sativum respond to gibberellin A(20) (GA(20)) in the dark, although the same dosage of GA(20) applied to light-grown le(5839) seedlings elicits no growth response. The acylcyclohexanedione growth retardant, LAB 198 999, which is known to inhibit gibberellin oxidation and in particular 3beta-hydroxylation such as the conversion of GA(20) to GA(1), also inhibits the growth response of dark-grown dwarf (le(5839)) seedlings to GA(20). Thus, the biological activity of GA(20) in the dark appears to be a consequence of its conversion to GA(1), even though it is known from studies with light-grown seedlings that the le mutation reduces the conversion of GA(20) to GA(1).

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Regulation of key enzymes of sucrose biosynthesis in soybean leaves : effect of dark and light conditions and role of gibberellins and abscisic Acid.

An important part in the understanding of the regulation of carbon partitioning within the leaf is to investigate the endogenous variations of parameters related to carbon metabolism. This study of diurnal changes in the activities of sucrose-synthesizing enzymes and levels of nonstructural carbohydrates in intact leaves of field-grown soybean plants (Glycine max [L.]) showed pronounced diurnal fluctuations in sucrose phosphate synthase (SPS) activity. However, there was no distinct diurnal change in the activity of fructose-1,6-bisphosphatase (F1,6BPase). SPS activity in leaves from plants grown in controlled environments presented two peaks during the light period. In contrast to field-grown plants, F1,6BPase activity in leaves from growth chamber-grown plants manifested one peak during the first half of the light period. In plants grown under both conditions, sucrose and starch accumulation rates were highest during early hours of the light period. By the end of the dark period, most of the starch was depleted. A pattern of diurnal fluctuations of abscisic acid (ABA) levels in leaves was also observed under all growing conditions. Either imposition of water stress or exogenous applications of ABA inhibited F1,6BPase activity. However, SPS-extractable activity increased following water deficit but did not change in response to ABA treatment. Gibberellin application to intact soybean leaves increased levels of both starch and sucrose. Both gibberellic acid (10(-6)m) and gibberellins 4 and 7 (10(-5)m) increased the activity of SPS but had an inconsistent effect on F1,6BPase. Correlation studies between the activities of SPS and F1,6BPase suggest that these two enzymes are coordinated in their function, but the factors that regulate them may be distinct because they respond differently to certain environmental and physiological changes.

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Gibberellin-like Substances from the Developing Banana Fruit.

The occurrence of 2 gibberellin-like substances was demonstrated in the developing banana fruit, Musa sapientum, Linn. Chemical and biological evidence led to the tentative identification of the 2 compounds as GA(7) and GA(x) (previously isolated from citrus fruits). Support for such identification was obtained from thin layer chromatography, gradient elution column chromatography, spectrofluorometry, the dwarf maize test, and the cucumber hypocotyl test. Significance of the GA(x) -designated compound increased since it is believed to occur in the fungus Fusarium moniliforme, Sheld. in addition to 2 different species of higher plants. It does not resemble any of the known gibberellins as far as chromatography is concerned.

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