Narrow-band light regulation of ethylene and gibberellin levels in hydroponically-grown <Emphasis Type="Italic">Helianthus annuus</Emphasis> hypocotyls and roots

Both hypocotyl and root growth of sunflower (Helianthus annuus) were examined in response to a range of narrow-band width light treatments. Changes in two growth-regulating hormones, ethylene and gibberellins (GAs) were followed in an attempt to better understand the interaction of light and hormonal signaling in the growth of these two important plant organs. Hydroponically-grown 6-day-old sunflower seedlings had significantly elongated hypocotyls and primary roots when grown under far-red (FR) light produced by light emitting diodes (LEDs), compared to narrow-band red (R) and blue (B) light. However, hypocotyl and primary root lengths of seedlings given FR light were still shorter than was seen for dark-grown seedlings. Light treatment in general (compared to dark) increased lateral root formation and FR light induced massive lateral root formation, relative to treatment with R or B light. Levels of ethylene evolution (roots and hypocotyls) and concentrations of endogenous GAs (hypocotyls) were assessed from both 6-day-old sunflower plants either grown in the dark, or treated with FR, R or B light. Both R and B light had similar effects on hypocotyl and root growth as well as on ethylene and on hypocotyl GA levels. Dark treatment resulted in the highest ethylene levels, whereas FR treatment significantly reduced ethylene evolution for both hypocotyls and roots. R- and B-light treatments elevated ethylene evolution relative to FR light. Endogenous GA₅₃ and GA₁₉ levels in hypocotyls were significantly higher and GA₄₄, GA₂₀ and GA₁ levels significantly lower, for dark and FR light treatments compared to R and B light-treatments. The patterns seen for changes in GA concentrations indicate FR-, R- and B-light-mediated effects [differences] in the metabolism of the early C₂₀ GAs, GA₅₃ → GA₄₄ → GA_19. Surprisingly, GA₂₀, GA₁ and GA₈ levels in hypocotyls were very much reduced by treatment of the plants with FR light, relative to B and R-light treatments, e.g. the increased hypocotyl elongation induced by FR light was correlated with reduced levels of all three of the downstream C₁₉ GAs. The best explanation, albeit speculative, is that a more rapid metabolism, i.e. GA₂₀ → GA₁ → GA₈ → GA₈ conjugates occurs under FR light. Although this study provided no evidence that elevated ethylene evolution by roots or hypocotyls of sunflower is controlling growth via endogenous GA biosynthesis, there are differences between soil-grown and hydroponically-grown sunflower seedlings with regard to trends seen for hypocotyl GA concentrations and both root and hypocotyl ethylene evolution in response to narrow band width R and FR light signaling.     

Responses of neotropical mangrove seedlings grown in monoculture and mixed culture under treatments of hydroperiod and salinity

We investigated the combined effects of salinity and hydroperiod on seedlings of Rhizophora mangle and Laguncularia racemosa grown under experimental conditions of monoculture and mixed culture by using a simulated tidal system. The objective was to test hypotheses relative to species interactions to either tidal or permanent flooding at salinities of 10 or 40 g/l. Four-month-old seedlings were experimentally manipulated under these environmental conditions in two types of species interactions: (1) seedlings of the same species were grown separately in containers from September 2000 to August 2001 to evaluate intraspecific response and (2) seedlings of each species were mixed in containers to evaluate interspecific, competitive responses from August 2002 to April 2003. Overall, L. racemosa was strongly sensitive to treatment combinations while R. mangle showed little effect. Most plant responses of L. racemosa were affected by both salinity and hydroperiod, with hydroperiod inducing more effects than salinity. Compared to R. mangle, L. racemosa in all treatment combinations had higher relative growth rate, leaf area ratio, specific leaf area, stem elongation, total length of branches, net primary production, and stem height. Rhizophora mangle had higher biomass allocation to roots. Species growth differentiation was more pronounced at low salinity, with few species differences at high salinity under permanent flooding. These results suggest that under low to mild stress by hydroperiod and salinity, L. racemosa exhibits responses that favor its competitive dominance over R. mangle. This advantage, however, is strongly reduced as stress from salinity and hydroperiod increase.     

Gibberellins and bud break,vegetative shoot growth and flowering in Metrosideros collina cv. Tahiti

Applications of the growth promotive gibberellins (GAs) GA₄ and 2,2-dimethyl GA₄, and of C-16,17 endo-dihydro GA₅, which is known to promote flowering while inhibiting stem growth in the long-day grass Lolium temulentum, were made to micropropagated plants of Metrosideros collina cv. Tahiti, a highly ornamental cultivar with an intermittent flowering pattern. Gibberellin A₄ and 2,2-dimethyl GA₄ stimulated vegetative growth both in elongating shoots, and internodes of shoots developing from buds that were quiescent at the time of GA application. Abscission of the apices of expanding shoots, a feature of mature Metrosideros plants, was inhibited by these GAs, the rejuvenation of micropropagated plantlets being enhanced. However, C-16,17 endo-dihydro GA₅ differed from GA₄ and 2,2-dimethyl GA₄ by having no promotive effects on vegetative growth, and no inhibition of apical abscission. Notwithstanding this contrasting effect on vegetative growth, high doses of GA₄ or C-16,17 endo-dihydro GA₅ similarly reduced flowering on shoots to which either GA was applied. Reduced flowering in response to applied GAs is common in many woody angiosperms, and in this instance was probably the combined result of abortion of developing floral structures in quiescent buds, and a preferential inhibition of bud break for floral buds relative to vegetative buds, particularly by GA₄. Finally, both C-16,17 endo-dihydro GA₅ and GA₄ strongly inhibited bud break in this woody angiosperm, although GA₄ could initially stimulate bud break when applied to vegetative buds close to the expansion stage. The above findings, in toto, highlight the sensitivity of Metrosideros to both classes of GA in a variety of growth and development processes.     

Gibberellin physiology of safflower: endogenous gibberellins and response to gibberellic acid

Endogenous gibberellins (GAs) were extracted from safflower (Carthamus tinctorius L.) stems and detected by capillary gas chromatography-mass spectrometry from which GA₁, GA₃, GA₁₉,, GA₂₀, GA₂₉, and probably, GA₄₄ were detected. The detection of these GAs suggests that the early 13-OH biosynthetic pathway is prevalent in safflower shoots. Deuterated GAs were used as internal standards and GA concentrations were determined in stems harvested at weekly intervals. GA₁ and GA₁₉ levels per stem increased but concentrations per gram dry weight decreased over time. GA₂₀ was only detected in young stem tissue.Gibberellic acid (GA₃) was also applied in field trials and both GA₃ and the GA biosynthetic inhibitor, paclobutrazol, were applied in growth chamber tests. GA₃ increased epidermal cell size, internode length, and increased internode cell number causing stem elongation. Conversely, paclobutrazol reduced stem height, internode and cell size, cell number and overall shoot weight. In field tests, GA₃ increased total stem weight, but decreased leaf weight, flower bud number and seed yield. Thus, GA₃ promoted vegetative growth at the expense of reproductive commitment. These studies collectively indicate a promotory role of GAs in the control of shoot growth in safflower, and are generally consistent with gibberellin studies of related crop plants. Author for correspondence     

Exophiala sp. LHL08 reprograms Cucumis sativus to higher growth under abiotic stresses

Endophytic fungi are potential sources of secondary metabolites; however, they are little known for phytohormones secretion and amelioration of plant growth under abiotic stresses. We isolated a novel endophyte from the roots of Cucumis sativus and identified it as a strain of Exophiala sp. by sequencing internal transcribed spacer/large subunit rDNA and phylogenetic analysis. Prior to identification, culture filtrate (CF) of Exophiala sp. has shown significant growth promotion of Waito‐C [a gibberellins (GAs)‐deficient mutant cultivar] and Dongjin‐byeo (normal GAs biosynthesis cultivar) rice seedlings. CF analysis of Exophiala sp. showed the presence of physiologically active GAs (GA₁, GA₃, GA₄ and GA₇) and inactive GAs (GA₅, GA₈, GA₉, GA₁₂ and GA₂₀). Exophiala sp. had higher GAs in its CF than wild‐type strain of Gibberella fujikuroi except GA₃. Influence of Exophiala sp. was assessed on cucumber plant's growth and endogenous abscisic acid (ABA), salicylic acid (SA) and bioactive GAs under salinity and drought stresses. Exophiala sp.‐treated plants have shown significantly higher growth and rescued the host plants from stress promulgated water deficit, osmotic and cellular damage. The altered levels of stress‐responsive ABA showed low level of stress confined to endophyte‐applied plants than control. Elevated levels of SA and bioactive GAs (GA₃ and GA₄) in endophyte‐associated plants suggest stress‐modulating response toward salinity and drought. In conclusion, symbiotic relations between Exophiala and cucumber have reprogrammed the host plant growth under abiotic stresses, thus indicating a possible threshold role of endophytic fungi in stress alleviation. This study could be extended for improving agricultural productivity under extreme environmental conditions.     

Floral initiation in sorghum hastened by gibberellic acid and far-red light

Combinations of far-red light (FR) (4 min) and gibberellic acid (GA₃), given at the beginning of a daily 12-h dark period in a growth room, were used to study floral induction in four maturity genotypes of the milo group of sorghum (Sorghum bicolor (L.) Moench). The 12-h dark period without GA₃ application or FR induced flowering in only the early genotype; FR hastened initiation in the early genotype, while GA₃ hastened floral initiation in the two intermidiate-flowering genotypes. GA₃ and FR together had a strong synergistic effect, hastening floral initiation by 30 to more than 80 d in the early and intermediate genotypes. Red light (R) did not hasten flowering; FR preceded by R gave the same effect as FR alone. GA₃ promoted stem elongation equally whether floral initiation occurred or not; thus, its effect on stem elongation was independent of floral initiation. The capacity of GA₃ to induce flowering in sorghum, a short-day plant, seems to be enhanced by phytochrome being in the P_R form at the beginning of the night when GA₃ was applied.Abbreviations FR far-red light - GA(s) gibberellin(s) - GA₃ gibberellic acid - R red light     

Promotive and inhibitory effects of uniconazole and prohexadione on the sprouting of bulbils of Chinese yam,Dioscorea opposita

Gibberellin A₁ (GA₁), GA₃ and GA₄ inhibited the sprouting of nondormant bulbils of Chinese yam, Dioscorea opposita, where the effectiveness of the GAs was as follows: GA₄>GA₁+GA₃. Uniconazole and prohexadione, plant growth retardants, promoted the sprouting of half-dormant bulbils. By contrast, these retardants inhibited the sprouting of nondormant bulbils. Gibberellin A₃ (GA₃) and A₄ (GA₄) which were applied to the stems of the sprouted bulbils, promoted stem elongation, but GAs applied to the bulbous parts inhibited this process. The effectiveness of the GAs on stem elongation was as follows: GA₃+GA₄ for the promotion and GA₄ > GA₃ for the inhibition. Uniconazole applied to the stem inhibited the stem elongation of the sprouted bulbils. These results suggest the possible involvement of endogenous GAs in the induction and maintenance of bulbil dormancy of D. opposita, as well as in the bulbil sprouting and subsequent stem elongation.     

The roles and interactions of ethylene with gibberellins in the far-red enriched light-mediated growth of <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> seedlings

Wild type (WT) tomato seedlings responded to a low red to far-red (R/FR) ratio with increased stem elongation, similar leaflet area expansion and lower shoot ethylene levels. The levels of endogenous growth-active GA₁ and its immediate precursor GA₂₀ were decreased by low R/FR ratio, whereas the levels of GA₁ catabolite, GA₈, increased. To examine the interaction of ethylene with GAs in regulating tomato shoot growth under low R/FR ratio, transgenic (T) seedlings bearing Le-ACS2 and Le-ACS4 antisense mRNA were utilized. Low R/FR ratio increased stem elongation and decreased ethylene levels in T tomato shoots, as it did in WT shoots. However, T stems were significantly taller than the WT stems under low R/FR ratio. Leaflet areas were significantly larger for T, than WT seedlings under both R/FR ratios. Low R/FR ratio did not decrease endogenous levels of GA₁ and GA₂₀ in T shoots, but did increase GA₈ levels, which were higher than in WT shoots. These results, and hormone/inhibitor application studies, showed that in tomato shoots subjected to low R/FR ratio, GAs play a growth-promotive role in stem elongation, whereas ethylene is growth-inhibitory. Further, these results may imply that decreasing ethylene production under low R/FR ratio causes increases in stem elongation and GA levels.     

Endophytic fungal pre-treatments of seeds alleviates salinity stress effects in soybean plants

In the present study, four endophytic fungi (GM-1, GM-2, GM-3, and GM-4) were tested for their ability to improve soybean plant growth under salinity stress conditions. The seed germination and plant growth were higher in seeds pretreated with endophytic fungal cultures than their controls. The positive influence of fungi on plant growth was supported by gibberellins analysis of culture filtrate (CF), which showed wide diversity and various concentrations of GAs. Specifically, GA₄, GA₇, GA₈, GA₉, GA₁₂, and GA₂₀ were found in fungal CFs. Under salinity stress conditions, GM-1 significantly enhanced the length and fresh weight of soybean plants relative to other fungal treatments. GM-1 effectively mitigated the adverse effects of salinity by limiting lipid peroxidation and accumulating protein content. GM-2, GM-3, and GM-4 also counteracted the salinity induced oxidative stress in soybean plants through reduction of lipid peroxidation and enhancement of protein content, maintaining the length and fresh weight of shoots. The activities of the antioxidant enzymes catalase, superoxide dismutase and peroxidase were inhibited in salinity exposed plants, while GM-1 significantly enhanced these antioxidant enzyme activities in plants under salt stress. GM-1 treatment also showed lower levels of abscisic acid and elevated levels of salicylic acid in plants under salinity stress. Hence, GM-1 was identified as Fusarium verticillioides (teleomorph Gibberella moniliformis) isolate RK01 based on its DNA sequence homology. These results suggest that endophytic fungal (F. verticillioides) pre-treatment of soybean seeds would be an effective method to promote soybean plant growth under salinity stress conditions.     

Gibberellins in seeds of Arabidopsis thaliana: biological activities,identification and effects of light and chilling on endogenous levels

The activities of several gibberellins in stimulating germination of wild-type and GA-deficient gal seeds of Arabidopsis thaliana were compared. Of the six compounds tested GA₄ and GA₇-isolactone had the highest activity and GA₇ and GA₉ the lowest; activities of GA₁ and GA₃ were intermediate. Combined application of pure GAs presented no indications that more than one GA receptor is involved. Four GAs were identified in extracts from wild-type and GA-insensitive gai seeds by combined gas chromatography mass spectrometry: GA₁, GA₃, GA₄ and GA₉. Effects of light and chilling on levels of GA₁, GA₄ and GA₉ were studied using deuterated standards. Light increased both GA levels and germination in unchilled wild-type and gai seeds. As a result of irradiation GA levels in gai seeds were 7–10 times as high as in wild-type seeds. In the dark germination was 0%, in the light 14% of gai seeds and 95% of wild-type seeds germinated. A chilling pre-treatment of 7 days at 2°C was required to enhance further the germination of gai seeds in the light. Light did not increase GA levels of chilled seeds of either genotype; levels of GA₄ and GA₉ of chilled gai seeds, in the light were respectively 7 and 12 times lower than in non-chilled seeds, whereas the latter seeds germinated better. Slightly elevated levels of GA₄ were detected in darkness after chilling, but germination capacity was still 0%. These results strengthened the conclusion that GAs are required for germination of A. thaliana seeds, whereby GA₄ has intrinsic biological activity. However, it is unlikely that light and chilling stimulate germination primarily by increasing levels of GA. Instead GA sensitivity is a possible alternative.     

Accumulation of C19-gibberellins in the gibberellin-insensitive dwarf mutantgai ofArabidopsis thaliana (L.) Heynh

The endogenous gibberellins (GAs) from shoots of the GA-insensitive mutant,gai, ofArabidopsis thaliana were analyzed and compared with the GAs from the Landsberg erecta (Ler) line. Twenty GAs were identified in Ler plants by full-scan gas chromatography-mass spectrometry (GC-MS) and Kovats retention indices (KRI's). These GAs are members of the early-13-hydroxylation pathway (GA₅₃, GA₄₄, GA₁₉, GA₁₇, GA₂₀, GA₁, GA₂₉, and GA₈), the non-3,13-hydroxylation pathway (GA₁₂, GA₁₅, GA₂₄, GA₂₅, GA₉, and GA₅₁), and the early-3-hydroxylation pathway (GA₃₇, GA₂₇, GA₃₆, GA₁₃, GA₄, and GA₃₄). The same GAs, except GA₅₃, GA₄₄, GA₃₇, and GA₂₉ were detected in thegai mutant by the same methods. In addition, extracts fromgai plants contained GA₄₁ and GA₇₁. Both lines also contained several unknown GAs. In Ler plants these were mainly hydroxy-GA₁₂ derivatives, whereas in thegai mutant hydroxy-GA₂₄, hydroxy-GA₂₅, and hydroxy-GA₉ compounds were detected. Quantification of seven GAs by GC-selected ion monitoring (SIM), using internal standards, and comparisons of the ion intensities in the SIM chromatograms of the other thirteen GAs, demonstrated that thegai mutant had reduced levels of all C₂₀-dicarboxylic acids (GA₅₃, GA₄₄, GA₁₉, GA₁₂, GA₁₅, GA₂₄, GA₃₇, GA₂₇, and GA₃₆). In contrast,gai plants had increased levels of C₂₀-tricarboxylic acid GAs (GA₁₇, GA₂₅, and GA₄₁) and of all C₁₉-GAs (GA₂₀, GA₁, GA₈, GA₉, GA₅₁, GA₄, GA₃₄, and GA₇₁) except GA₂₉. The 3β-hydroxylated GAs, GA₁ and GA₄, and their respective 2β-hydroxylated derivatives, GA₈ and GA₃₄, were the most abundant GAs found in shoots of thegai mutant. Thus, thegai mutation inArabidopsis results in a phenotype that resembles GA-deficient mutants, is insensitive to both applied and endogenous GAs, and contains low levels of C₂₀-dicarboxylic acid GAs and high levels of C₁₉-GAs. This indicates that theGAI gene controls a step beyond the synthesis of an active GA. Thegai mutant is presumably a GA-receptor mutant or a mutant with a block in the transduction pathway between the receptor and stem elongation. We thank Dr. L.N. Mander, Australian National University, Canberra, for providing [²H]gibberellins, Dr. B.O. Phinney, University of California, Los Angeles, USA for [¹³C]GA₈, and Dr. D.A. Gage, MSU-NIH Mass Spectrometry Facility (grant No. DRR00480), for advice with mass spectrometry. This work was supported by a fellowship from the Spanish Ministry of Agriculture (I.N.I.A.) to M.T., by the U.S. Department of Energy under Contract DE-ACO2-76ERO-1338, and by U.S. Department of Agriculture grant No. 88-37261-3434 to J.A.D.Z.     

Gibberellin Metabolism and Transport During Germination and Young Seedling Growth of Pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

The role of gibberellins (GAs) during germination and early seedling growth is examined by following the metabolism and transport of radiolabeled GAs in cotyledon, shoot, and root tissues of pea (Pisum sativum L.) using an aseptic culture system. Mature pea seeds have significant endogenous GA₂₀ levels that fall during germination and early seedling growth, a period when the seedling develops the capacity to transport GA₂₀ from the cotyledon to the shoot and root of the seedling. Even though cotyledons at 0–2 days after imbibition have appreciable amounts of GA₂₀, the cotyledons retain the ability to metabolize labeled GA₁₉ to GA₂₀ and express significant levels of PsGA20ox2 message (which encodes a GA biosynthesis enzyme, GA 20-oxidase). The large pool of cotyledonary GA₂₀ likely provides substrate for GA₁ synthesis in the cotyledons during germination, as well as for shoots and roots during early seedling growth. The shoots and roots express GA metabolism genes (PsGA3ox genes which encode GA 3-oxidases for synthesis of bioactive GA₁, and PsGA2ox genes which encode GA 2-oxidases for deactivation of GAs to GA₂₉ and GA₈), and they develop the capacity to metabolize GAs as necessary for seedling establishment. Auxins also show an interesting pattern during early seedling growth, with higher levels of 4-chloro-indole-3-acetic acid (4-Cl-IAA) in mature seeds and higher levels of indole-3-acetic acid (IAA) in young root and shoot tissues. This suggests a changing role for auxins during early seedling development.     

Over-expression of a gibberellin 2-oxidase gene from Phaseolus coccineus L. enhances gibberellin inactivation and induces dwarfism in Solanum species

Gibberellins (GAs) are endogenous hormones that play a predominant role in regulating plant stature by increasing cell division and elongation in stem internodes. The product of the GA 2-oxidase gene from Phaseolus coccineus (PcGA2ox1) inactivates C₁₉-GAs, including the bioactive GAs GA₁ and GA₄, by 2β-hydroxylation, reducing the availability of these GAs in plants. The PcGA2ox1 gene was introduced into Solanum melanocerasum and S. nigrum (Solanaceae) by Agrobacterium-mediated transformation with the aim of decreasing the amounts of bioactive GA in these plants and thereby reducing their stature. The transgenic plants exhibited a range of dwarf phenotypes associated with a severe reduction in the concentrations of the biologically active GA₁ and GA₄. Flowering and fruit development were unaffected. The transgenic plants contained greater concentrations of chlorophyll b (by 88%) and total chlorophyll (11%), although chlorophyll a and carotenoid contents were reduced by 8 and 50%, respectively. This approach may provide an alternative to the application of chemical growth retardants for reducing the stature of plants, particularly ornamentals, in view of concerns over the potential environmental and health hazards of such compounds. C. Dijkstra, E. Adams, A. Bhattacharya and A. F. Page contributed equally to this paper.     

Stem elongation and gibberellins in alpine and prairie ecotypes of Stellaria longipes

The potential for gibberellins (GAs) to control stem elongation and itsplasticity (range of phenotypic expression) was investigated inStellaria longipes grown in long warm days. Gibberellinmetabolism and sensitivity was compared between a slow-growing alpine dwarfwithlow stem elongation plasticity and a rapidly elongating, highly plastic prairieecotype. Both ecotypes elongated in response to exogenous GA₁,GA₄ or GA₉, but surprisingly, the alpine dwarf wasrelatively unresponsive to GA₃. Endogenous GA₁,GA₃, GA₄, GA₅, GA₈, GA₉and GA₂₀ were identified and quantified in stem tissue harvested atcommencement, middle and end of the period of most rapid elongation. Theconcentration of GAs which might be expected to promote shoot elongation washigher during rapid elongation than toward its end for both ecotypes. Whilethere was a trend for certain GAs (GA₃, GA₄,GA₉, GA₂₀) to be higher in stems of the alpine ecotypeduring rapid elongation, that result does not explain the slower growth of thealpine ecotype and the faster growth of the prairie ecotype under a range ofconditions. To determine if the two ecotypes metabolized GA₂₀differently, plants were fed [²H]- or[³H]-GA₂₀. The metabolic products identified included[²H₂]-GA₁, -GA₈, -GA₂₉,-GA₆₀, -3-epi-GA₁, GA₁₁₈(-1-epi-GA₆₀) and -GA₇₇. The concentration of[²H₂]-GA₁ also did not differ between the twoecotypes and metabolism of [²H₂]- or[³H]-GA₂₀ was also similar. In the same experiments thepresence of epi-GA₁, GA₂₉, GA₆₀,GA₁₁₈ and GA₇₇ was indicated, suggesting that these GAsmay also occur naturally in S. longipes, in addition tothose described above. Collectively, these results suggest that while stemelongation within ecotypes is likely regulated by GAs, differences in GAcontent, sensitivity to GAs (GA₃ excepted), or GA metabolism areunlikely to be the controlling factor in determining the differences seen ingrowth rate between the two ecotypes under the controlled environmentconditionsof this study. Nevertheless, further study is warranted especially underconditions where environmental factors may favour a GA:ethylene interaction.     

Transport and metabolism of exogenously applied gibberellins to Malus domestica Borkh. cv. Jonagold

The radio-labeled gibberellins GA₁, GA₃,GA₄, and GA₇ were applied to intact developing applefruits (Malus domestica Borkh. cv. Jonagold) during theperiod when GAs are suggested to inhibit flower bud induction for the followingyear. Radioactivity from these compounds was found to be transported intoadjacent tissues as there are pedicels and bourses (4%). Application topedicels, after removal of the fruits, enhanced the transport into adjacentbourses up to 11%. The bud-carrying lateral bourse shoots contained onlyminor amounts of radioactivity on average 0.4% in both cases. Theseexport rates were identical, 1 or 5 days after application.After application of the corresponding deuterium-labeled GAs and analyses bymass spectrometry the specific metabolization of GA₁ toGA₁ 13-O-glucoside and of GA₃ to GA₃13-O-glucoside was demonstrated. Additional metabolites of GA₁ andGA₃ were not detected. After fruit application of GA₃ theratio of GA₃ to GA₃ 13-O-glucoside was found to be 1:2 inthe fruit. Pedicel application led to ratios of 1:4 and 1:5, respectively, inthe pedicel and in the adjacent bourse. After the application of GA₄and GA₇, neither glucosylation products nor other GA-like metabolitescould be identified.This is the first report of the metabolism of GAs to GA 13-O-glucosides indeveloping apple fruits. The possible function of the GAs as a signal in flowerbud formation for the following year is discussed.     

Endogenous Gibberellins in the Developing Liquid Endosperm of Tea

Changes in the kind and level of endogenous gibberellins (GAs) in the developing liquid endosperm of tea (Camellia sinensis L.) were investigated. Gibberellin A₁ (GA₁), GA₈, GA₁₉, GA₂₀, and GA₄₄ were identified by GC-MS or GC-SIM. Besides these early C-13 hydroxylated GAs, GA₃, iso-GA₃, and GA₃₈ were also identified. Of these GAs, GA₁ and GA₃ were the major gibberellins. The levels of these GAs were at a maximum in the globular embryo stage and then decreased rapidly during embryo maturation.     

Interaction of Brassinosteroids with Light Quality and Plant Hormones in Regulating Shoot Growth of Young Sunflower and <Emphasis Type="Italic">Arabidopsis</Emphasis> Seedlings

Sunflower hypocotyls elongate as light quality changes from the normal red to far-red (R/FR) ratio of sunlight to a lower R/FR ratio. This low R/FR ratio-induced elongation significantly increases endogenous concentrations of indole-3-acetic acid (IAA) and also of three gibberellins (GAs): GA₂₀, GA₁, and GA₈. Of these, it is likely GA₁ that drives low R/FR-induced growth. Brassinosteroids are also involved in shoot growth. Here we tested three R/FR ratios: high, normal, and low. Significant hypocotyl elongation occurred with this stepwise reduction in R/FR ratio, but endogenous castasterone concentrations in the hypocotyls remained unchanged. Brassinolide was also applied to the seedlings and significantly increased hypocotyl growth, though one that was uniform across all three R/FR ratios. Applied brassinolide increased hypocotyl elongation while significantly reducing (usually) levels of IAA, GA₂₀, and GA₈, but not that of GA₁, which remained constant. Given the above, we conclude that endogenous castasterone does not mediate the hypocotyl growth that is induced by enriching FR light, relative to R light. Similarly, we conclude that the hypocotyl growth that is induced by applied brassinolide does not result from an interaction of brassinolide with changes in light quality. The ability of applied brassinolide to influence IAA, GA₂₀, and GA₈ content, yet have no significant effect on GA₁, is hard to explain. One speculative hypothesis, though, could involve the brassinolide-induced reductions that occurred for endogenous IAA, given IAA’s known ability to differentially influence the expression levels of GA20ox, GA3ox, and GA2ox, key genes in GA biosynthesis.     

Gibberellins and the photoperiodic control of leaf growth in Poa pratensis

The role of gibberellins (GAs) in photoperiodic control of leaf elongation in Poa pratensis was studied by both application of exogenous GAs and analysis of endogenous GAs. Leaf elongation was strongly increased under long day (LD, 24 h) conditions at both 9 and 21°C, leaf length at 9°C LD being similar to that in plants grown in short days (SD, 8 h) at 21°C. However, even at 21°C leaf elongation was enhanced by LD. Exogenous GA₁ could completely compensate for LD at both 9 and 21°C. Gibberellins A₂₀, A₁₉ and A₄₄ could also partly replace LD, but they were significantly less active than GA₁, GA₅₃ was inactive when applied to plants grown at 9°C in SD and exhibited only marginal activity at 9°C LD and 21°C SD. The total level of GAs of the early 13-hydroxylation pathway (A₅₃, A₄₄, A₁₉, A₂₀ and A₁) increased rapidly when plants were transferred from SD to LD at 9°C. After transfer from 9 to 21°C, there was an increase in GA levels at both LD and SD, followed by a decrease under LD conditions. In all cases, GA₁₉ was the predominant GA, accounting for 60 to 80% of the analysed GAs. Levels of the bioactive GA₁ were low and increased transiently by LD four days after transfer from SD to LD. At both temperatures, the ratio GA₁₉ to GA₂₀ and GA₂₀ to GA₁ at 9°C was enhanced by LD compared with SD. Taken together, these results support the hypothesis that photoperiodic regulation of leaf elongation in Poa pratensis is GA-mediated, and they indicate a photoperiodic control of oxidation of GA₅₃ to GA₄₄ and GA₁₉ to GA₂₀, and perhaps also of 3β-hydroxylation of GA₂₀ to GA₁.     

Environmental and hormonal regulation of seed dormancy and germination in Cape fynbos Leucospermum R.Br. (Proteaceae) species

The endogenous levels of abscisic acid (ABA), cytokinins (CKs) and gibberellins (GA₁/GA₃ combined) in Leucospermum glabrum embryos were monitored in axes and cotyledons separately during normal germination. Plant growth substance changes were correlated with known morphological, structural and ultrastructural events in the embryo of Proteaceae. The effect of exogenous application of 6-benzyladenine (BA) and GA₄₊₇ under three known dormancy-enforcing environmental conditions were studied in L. glabrum and L. cordifolium. The endogenous levels of the hormone classes GAs and CKs changed phasically during normal germination under a single alternating temperature regime. GA₁/GA₃ levels increased in cotyledons within 3 d of hydration while at the same time initial CK levels decreased. Following this transient peak GAs fell to a low level throughout the germinative period. Subsequently the CKs, Z and ZR, and to a lesser extent their dihydro-derivatives, appeared in both the axes and the cotyledons as fluctuating, transient peaks. Early increases in GAs are thought to control the induction of the germination process. The CK pattern suggests that CKs control at least three major processes of germination sensu stricto following induction: 1) early mobilization of protein and lipid reserves in the axis and later in cotyledons, 2) cotyledon expansion which causes the endotesta to split permitting radicle protrusion and 3) later, radicle growth.Our results indicate that dormancy in intact Leucospermum seeds is enforced by embryo anoxia, regulated by the impermeable exotesta. In addition synthesis of or tissue sensitizing to both hormone classes GAs and CKs depends on moderately low temperature as the primary environmental requirement. For GA synthesis a secondary, daily pulse of high temperature is required. Inhibitory hormones, specifically ABA, appear not to play a role.Abbreviations ABA Abscisic acid - BA 6-benzyladenine - CK Cytokinin - DHZ Dihydrozeatin - DHZR Dihydrozeatin riboside - GA Gibberellin - HPLC High performance liquid chromatography - iP Isopentenyladenine - IPA Isopentenyladenosine - PGS Plant growth substance - RIA Radioimmunoassay - Z Zeatin - ZR Zeatin riboside     

Endogenous gibberellins in flushing buds of three deciduous trees: Alder,aspen, and birch

Endogenous gibberellins (GAs) were extracted from flushing (expanding) vegetative buds of river alder (Alnus tenuifolia), European white birch (Betula pendula), and aspen (Populus tremuloides) and identified by gas chromatography-mass spectrometry with full scans and/or selected ion monitoring. Five 13-hydroxylated GAs were detected from the three trees: GA_{1, 8}, and ₂₀ from alder, GA_{1, 8, 19} and ₂₀ from aspen and GA_{1, 8, 19, 20}, and ₂₉ from birch. Thirteen other GAs previously detected in Salix or common in other plants were specifically investigated but not detected. The presence of GA₁, its probable precursors GA₁₉ and GA₂₀, and its probable metabolite, GA₈, suggests that the early 13-hydroxylated GA biosynthetic pathway is dominant in vegetative buds of these trees. Abundant endogenous GAs of these trees are similar to the principal GAs of willows (various Salix spp.) and poplars (various Populus spp.). This suggests similarities in the GA physiology and is consistent with a common role of GA₁ as a regulator of shoot growth in woody angiosperms.