Effect of gibberellin biosynthesis inhibitors on shoot regeneration from hypocotyl explants of Albizzia julibrissin

Summary Hypocotyl explants of Albizzia julibrissin were placed on Gamborg's B5 medium supplemented with various levels of paclobutrazol, uniconazole, prohexadione calcium, or GA₃. Callus formation was evident within one week after placement of the explants on the culture media. Green nodule-like structures protruded from the distal end of the explants within 10 days and developed into shoots within a month. These shoots readily formed adventitious roots when placed on fresh culture medium. All three of the gibberellin biosynthesis inhibitors increased shoot formation compared to the control. The number of shoots per explants was increased 107, 79, and 168% by 0.3–0.4 μM paclobutrazol, uniconazole, and prohexadione calcium, respectively. In contrast to the gibberellin biosynthesis inhibitors, GA₃ decreased shoot formation. These results indicate that modification of gibberellin status can have a strong impact on the number of shoots formed.     

Thermomorphogenic and Photomorphogenic Control of Stem Elongation in Fuchsia Is Not Mediated by Changes in Responsiveness to Gibberellins

Stem elongation in Fuchsia × hybrida was influenced by cultivation at different day and night temperatures or in different light qualities. Internode elongation of plants grown at a day (25°C) to night (15°C) temperature difference (DIF+10) in white light was almost twofold that of plants grown at the opposite temperature regime (DIF−10). Orange light resulted in a threefold stimulation of internode elongation compared with white light DIF−10. Surprisingly, internode elongation in orange light was similar for plants grown at DIF−10 and DIF+10. Flower development was accelerated at DIF−10 compared with DIF+10 in both white and orange light. To examine whether the effects of DIF and light quality on shoot elongation were related to changes in gibberellin metabolism or plant sensitivity to gibberellins (GAs), the stem elongation responses of paclobutrazol-treated plants to applied gibberellins were determined. In the absence of applied gibberellins paclobutrazol (>0.32 μmol plant⁻¹) strongly retarded shoot elongation. This inhibition was nullified by the application of about 10–32 nmol of GA₁, GA₄, GA₉, GA₁₅, GA₁₉, GA₂₀, GA₂₄, or GA₄₄. The results are discussed in relation to possible effects of DIF and light quality on endogenous gibberellin levels and gibberellin sensitivity of fuchsia and their effects on stem elongation. Received October 4, 1997; accepted December 17, 1997     

A study on floral biology of seedlings in vitro in Orychophragmus violaceus: Induction of flowers in seedlings of O. violaceus cultured in vitro

Seedlings of O. violaceus were cultured on MS media and treated at low temperature. Cold treatment at 5–7 °C for more than 7 days was needed for flower induction of seedlings in vitro originated from germinated seeds. When cultured on MS media supplemented with 2.5 mg l⁻¹ zeatin and 2 mg l⁻¹ gibberellin (GA₃), seedlings in vitro did initiate flowers without cold treatment. When MS media was used with a reduced amount of NH₄NO₃, flower induction of seedlings in vitro could be accelerated. This revised version was published online in June 2006 with corrections to the Cover Date.     

Gibberellin A1 dwarfism and shoot elongation in higher plants

Evidence is presented to support the generalization that gibberellin A₁ is the main ‘gibberellin hormone’ that is activeper se in the control of elongation growth in higher plants. The evidence is based on a combination of chemical and genetic studies using single gene mutants of maize, pea and rice.     

The effects of IAA and tetcyclacis on tuberization in potato (Solanum tuberosum L.) shoot cultures in vitro

Potato (Solanum tuberosum cv. Désirée) shoots grown in vitro in continuous darkness or in long days (LDs), were used to investigate indole-3-acetic acid (IAA) effects on stolon initiation and tuber formation, combining IAA with increased or decreased gibberellin levels. An increased gibberellin (GA) level was achieved by the applying 1 μM GA₃, while decreased gibberellin level was presumably realized by the adding 3 μM tetcyclacis (Tc). About 15% of potato shoots developed stolons both in LDs and in darkness. Stolon initiation was stimulated by GA₃ in darkness and by Tc in LDs. Tuber formation was strongly inhibited in LDs and by GA₃ both in light and darkness, but stimulated in darkness at low GA level. Exceptionally, tuber formation occurred in LDs at the highest Tc concentrations, in about 25% of explants. Indole-3-acetic acid alone stimulated stolon formation in LDs, both in the presence or absence of GA₃. IAA alone also stimulated tuber formation in dark-grown shoots, but could not overcome the inhibitory effect of LDs. Indications that, depending on their concentration ratio, IAA may interact with GA₃ in different tuberization phases, have been discussed. Radomir Konjević—Deceased in July 2006.     

Inhibitors of animal phospholipase A2 enzymes are selective inhibitors of auxin-dependent growth. Implications for auxin-induced signal transduction

Auxin and elicitors reportedly activate phospolipase A. A number of inhibitors known to inhibit animal phospholipase A₂ were tested for their ability to inhibit hormone and fusicoccin-induced growth. To this end, growth induced by indolyl-3-acetic acid and 2,4-dichlorophenoxyacetic acid in hypocotyl segments of etiolated zucchini (Cucurbita pepo L.) seedlings was determined in the presence of the inhibitors nordihydroguajaretic acid (NDGA), aristolochic acid, 5,8,11,14-eicosatetraynoic acid (ETYA), PB_x (a prostaglandin derivative), and oleylethyl phosphocholine. Each chemical proved inhibitory to auxin-induced growth, oleylethyl phosphocholine being the least effective. The effects of the first three inhibitors were investigated in more detail. Growth induced by 10 μM 2,4-dichlorophenoxyacetic acid or 1 μM indolyl-3-acetic acid was inhibited 50% by about 30–50 μM NDGA, by about 25 μM aristolochic acid, and by about 10–20 μM EYTA. Growth inhibition was reversible and became apparent 0.5–1 h after inhibitor addition. Growth induced by 0.5 or 1 μM fusicoccin was much less inhibited by NDGA and by ETYA, whereas aristolochic acid was only slightly less effective on fusicoccin-induced than on auxin-induced growth. These three inhibitors were also tested for their effects on gibberellin-induced growth in light-grown peas (Pisum sativum L.) and on cytokinin-induced expansion growth in excised cotyledons from radish (Raphanus sativum L.) seedlings. In both tests, aristolochic acid had toxic side-effects although gibberellin-induced growth was still apparent. In the gibberellin test, neither NDGA at up to 100 μM nor ETYA at 80 μM was inhibitory to hormone-induced growth. Moreover, 40 μM ETYA was not inhibitory to kinetin-induced growth. We hypothesize that the selectivity of phospholipase A₂ inhibitors for auxin-induced growth implies a different signal transduction pathway for each of the different signal substances tested, and that auxins might use fatty acid(s) and/or lysophospholipid(s) or their derivatives as the preferred second messengers. Received: 24 September 1996 / Accepted: 18 January 1997     

Comparison of the effects of temperature and water activity on growth rate of food spoilage moulds

The influence of temperature (T) and water activity (a _w) on the growth rate (μ) of seven moulds (Alternaria alternata, Aspergillus flavus, Cladosporium cladosporioides, Mucor racemosus, Penicillium chrysogenum, Rhizopus oryzae and Trichoderma harzianum) was assessed in suboptimal conditions. Firstly, the dependence of fungal growth on temperature, at a _w 0.99, was modelled through an approach described previously for bacteria. A dimensionless growth rate variable: μ _dimα=μ/μ _optα depended on the following normalised temperature: T _dim=(T−T _min)/(T _opt− T _min) according to a power function: μ _dimα=[T _dim] ^α , where α was an exponent to be estimated. Secondly, the same approach was used to describe the influence of a _w on fungal growth, at the respective optimum temperatures for each mould. Similarly, μ _dimβ=μ/μ _optβ depended on the following normalised water activity: a _wdim=(a _w−a _wmin)/(a _wopt−a _wmin) according to a power function: μ _dimβ=[a _wdim]^β. Results show: (i) for each mould, the α-value is significantly less than the β-value, confirming that water activity has a greater influence than temperature on fungal development; (ii) the α-values and the β-values depend on the mould; (iii) the α-value is less than 1 for the mesophilic mould A. flavus, whereas the other moulds are characterised by higher α-values ranging from 1.10 to 1.54; (iv) the mesophilic A. flavus exhibits a low β-value, 1.50, compared to the hydrophilic T. harzianum, β=2.44, while β-values are within the range (1.71–2.37) for the other moulds. Journal of Industrial Microbiology & Biotechnology (2002) 28, 311–315 DOI: 10.1038/sj/jim/7000248 Received 27 June 2001/ Accepted in revised form 04 February 2002     

Effect of triterpenoid glycosides on α- and β-amylase activity and total protein content in wheat seedlings

Influence of the aleanolic acid glycosides from Silphium perfoliatum L. (silphioside B, C, E and G) and their progenins on the amylase activity and total protein content in wheat seedlings was studied. Treatment of the Triticum aestivum L. seeds with 1–10 μM water solutions of mono- and diglycosides (mono- and bisdesmosines) elevated the α-amylase and total amylase activities in seedlings. Silphioside E containing three glucose moieties in its molecule did not change α-amylase activity, but it did if bis-triglycoside acetylated carbohydrate (as in silphioside C). Effects of 5–10 μM solutions of the active glycosides was comparable with that of exogenous gibberellin A₃ and 6-benzylaminopurine.     

Evaluation of the maximum specific growth rate of a yeast indicating non-linear growth trends in batch culture

The maximum specific growth rate (μ_max) of an ethanolic D-xylose-fermenting yeast, Pichia stipitis, showing non-linear growth trends in batch culture, was calculated using the rate equation μ₂ = (1/Δt) ln(x ₂/x ₁). The absolute error Δμ, affecting μ₂, was derived using an equation given by Borzani (1994). Based on the assumption of linearity of growth curves between two closest time points, the relation between the two rate formulae, μ₁ = (1/xˉ)dx _t /dt and μ₂ = (1/Δt) ln(x ₂/x ₁) was established. In a particular condition, when μ₁ = μ₂, an equation has been developed, the roots of which are the specific growth rates at different time points. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ethylene-induced stem growth of deepwater rice is correlated with expression of gibberellin- and abscisic acid-biosynthetic genes

Ethylene decreases the content of endogenous abscisic acid (ABA) and increases the level of bioactive gibberellin A₁ (GA₁) in the submerged internodes of deepwater rice. During partial submergence, internodes of deepwater rice undergo rapid elongation as a result of ethylene accumulation in the internodal lacunae. In anin vitro experiment using stem sections from deepwater rice, treatment with 5 μL L^-1 ethylene promoted stem growth by up to 3.2-foId times over air treatment. Expression patterns were analyzed for genes that encode GA- and ABA-biosynthesis enzymes to determine any possible molecular basis for the changes observed in GA₁ and ABA contents as a result of ethylene action. Expression of theOsGA20ox2 andOsGA20ox4 genes, which encode GA 20-oxidase, and of theOsGA3ox2 gene, which encodes the enzyme that converts GA₂₀ to CA₁, was up-regulated, whereas that of three ABA-biosynthetic genes —OsNCED1, OsNCED2, andOsNCEDS-was down-regulated in the presence of ethylene. These results indicate that GA and ABA contribute equally to the submergence-or ethylene-induced stem elongation of deepwater rice via the coordinated and opposite regulation of biosynthesis.     

Gibberellin and ethylene control endoreduplication levels in the Arabidopsis thaliana hypocotyl

We have previously shown that endoreduplication levels in hypocotyls of Arabidopsis thaliana (L.) Heynh. are under negative control of phytochromes. In this study, the hormonal regulation of this process was analysed using a collection of A. thaliana mutants. The results show that two hormones in particular, gibberellin (GA) and ethylene, play distinct roles. Hypocotyl cells of the GA-deficient mutant ga1-11 grown in the dark did not elongate and showed a greatly reduced endoreduplication. Normal endoreduplication could be restored by supplying 10⁻⁹ M of the gibberellin GA₄₊₇, whereas the restoration of normal cell growth required 100-fold higher concentrations. The GA-insensitive mutant gai showed reduced cell elongation but normal ploidy levels. We conclude that (i) GA₄₊₇ has a global positive effect on endoreduplication and (ii) that endoreduplication is more sensitive to GA₄₊₇ than cell elongation. Ethylene had a completely different effect. It induced an extra round of endoreduplication both in light- and dark-grown seedlings and acted mainly on discrete steps rather than having a global effect on endoreduplication. The genes EIN2 and CTR1, components of the ethylene signal transduction pathway were both involved in this process. Received: 27 February 1999 / Accepted: 21 May 1999     

Dimensionless analysis of the microbial growth rate dependence on sub-optimal temperatures

The influence of sub-optimal temperatures (T) on the microbial growth rate (μ) has been assessed by means of dimensionless variables: T_dim = [T−T_min]/[T_opt−T_min] and μ_dim = μ/μ_opt. T_min represents the temperature at which there is no growth, T_opt the optimum temperature at which the growth rate, μ_opt, is maximum. Data sets, growth rate vs temperature, have been taken from the literature for 12 organisms (psychrotrophs, mesophiles and thermophiles). In order to compare these organisms, the power law function has been used: [μ_dim] = [T_dim]^α. The parameters μ_opt and T_opt are determined from direct readings whereas T_min and αare estimated by means of a non-linear regression. An accurate estimation of T_min is obtained providing low growth rate data are available. A wide range of optimal temperatures where the growth rate almost equals μ_opt prevents one from obtaining a narrow confidence interval forα. On the basis of the analysis hereafter developed, thermophiles are characterized by values of the power α less than mesophiles and psychrotrophs. Almost all of these values are significantly different from two, previously determined for Staphylococcus xylosus and widely used for predicting the microbial growth in foods. Received 15 May 1998/ Accepted in revised form 25 September 1998     

The Effects of Gibberellin4+7 on the Vase Life and Flower Quality of Alstroemeria Cut Flowers

Leaf yellowing is a major problem in Alstroemeria and absence of leaf senescence symptoms is an important quality attribute. Two Alstroemeria cultivars ‘Yellow King’ and ‘Marina’ were sourced from a commercial farm and harvested when sepals began to reflex. Stems were re-cut under water and kept in vase solutions of gibberellin A₄₊₇ (0, 2.5, 5.0, 7.5, 10.0, 12.5 or 15.0 mg l⁻¹ [Provide^r]). Treatments and cultivars were combined in a factorial fashion and arranged in a completely randomised design. Application of GA₄₊₇ in the holding solution at 2.5–10.0 mg l⁻¹ significantly delayed the onset of leaf senescence by around 7 days and significantly increased days to 50% petal fall by ca. 2 days. Additionally, these GA₄₊₇ concentrations resulted in higher retention of leaf nitrogen, leaf chlorophyll and also increased leaf water content, while reducing leaf dry weight, all relative to untreated controls. Cultivar ‘Yellow King’ had significantly longer vase life and a better retention of leaf quality than ‘Marina’. Our results suggest that a concentration of 10 mg l⁻¹ GA₄₊₇ can be used to prolong vase life, delay leaf senescence and enhance post-harvest quality of Alstroemeria cut flowers during their transport to market.     

Dependence of cell yield and specific growth rate ofCandida boidinii on temperature during continuous cultivation

Investigation of the dependence of cell yield ofCandida boidinii in a chemostat and specific growth rate in a turbidostat on temperature was carried out using methanol as substrate. Each of the curves obtained had only one maximum, the optimum temperature for yield (Y _s )and for specific growth rate (μ) being somewhat different. With deviation of temperature from the corresponding optimum the μ value decreases more sharply than that ofY _s . The shape of two-factor dependencesY _s (μ, T) and μ(S, T) is analyzed on the basis of these data and the known dependences ofY _s on μ and μ on the limiting substrate concentrationS. It is shown that the decelerating action of methanol on the growth rate increases at elevated and lower temperatures.     

The effect of growth conditions on the biodegradation of tributyl phosphate and potential for the remediation of acid mine drainage waters by a naturally-occurring mixed microbial culture

The biodegradation of tributyl phosphate (Bu₃-P, TBP), releasing phosphate at a high enough concentration locally to precipitate uranium from solution, was demonstrated by a mixed culture consisting primarily of pseudomonads. The effect of various parameters on Bu₃-P biodegradation by growing cells is described. Growth at the expense of Bu₃-P as the carbon and phosphorus source occurred over a pH range from 6.5 to 8, and optimally at pH 7. Bu₃-P biodegradation was optimal at 30 °C, reduced at 20 °C and negligible at 4 °C and 37 °C. Incorporation of Cu or Cd inhibited, and Ni, Co and Mn reduced its degradation. Inorganic phosphate (above 10 mM) and kerosene (up to 1 g/l) reduced Bu₃-P biodegradation significantly, but nitrate had no effect. Sulphate (10–100 mM) was inhibitory. When pregrown biomass was used the fastest rates of tributyl and dibutyl phosphate biodegradation were 25 μmol h⁻¹ mg protein⁻¹ and 37 μmol h⁻¹ mg protein⁻¹ respectively. Microcarrier-immobilised biomass decontaminated uranium-bearing acid mine waste water by uranium phosphate precipitation at the expense of Bu₃-P hydrolysis in the presence of 35 mM SO₄ ²⁻. At pH 4.5, 79% of the UO₂ ²⁺ was removed at a flow rate of 1.4 ml/h on a 7-ml test column. Received: 2 June 1997 / Received revision: 15 September 1997 / Accepted: 19 September 1997     

A novel method for characterisation of microbial growth kinetics on volatile organic compounds

A novel method for the determination of microbial growth kinetics on hydrophobic volatile organic compounds (VOC) has been developed. A stirred tank reactor was operated as a fed-batch system to which the VOC was continuously fed via the gas phase, assuring a constant VOC concentration in the mineral medium. A flow of air was saturated with the VOC, and then mixed with a further flow of air, to obtain a predetermined VOC concentration. Thus, different VOC concentrations in the mineral medium could be obtained by altering the VOC concentration in the feed gas. The growth kinetics of Xanthobacter autotrophicus GJ10 on 1,2-dichloroethane (DCE) and of Pseudomonas sp. strain JS150 on MonoChloroBenzene (MCB) were assessed using this method. The growth of strain JS150 was strongly inhibited at MCB concentrations higher than 160 mg l⁻¹, and the results were fitted using a piecewise function. The growth kinetics of strain GJ10 were described by the Luong model where maximum growth rate μ_max = 0.12 h⁻¹, substrate saturation constant K _S = 7.8 mg l⁻¹, and maximum substrate concentration S _m (above which growth is completely inhibited) = 1080 mg l⁻¹. Varying nitrogen and oxygen flows enabled the effect of oxygen concentration on the growth kinetics of Pseudomonas JS150 to be determined. Received: 30 November 1998 / Received revision: 19 March 1999 / Accepted: 20 March 1999     

In vitro propagation of caper (Capparis spinosa L.)

Summary A twenty fold multiplication per twenty days of caper was achieved by culturing nodal shoot segments in the presence of BAP (4 μM) plus IAA (0.3 μM) and GA₃ (0.3 μM). The use of a modified MS medium facilitated this response. Plantlet regeneration was induced on single shoots taken from proliferating clusters subcultured for 20 days on a reduced BAP (2 μM) without auxin and gibberellin Higher rooting responses (70%) were obtained after a 20-day incubation period in darkness on solid half-strength MS1 medium plus IAA (30 μM), followed by a subsequent 20 day culture period on half-strength MSI basal medium. Proliferation was mainly due to axillary shoot-bud development as revealed by histological studies. The extensive meristematic activities observed indicated the enormous morphological potential of this species.     

Regulation of Medicago sativa L. somatic embryos regeneration by gibberellin A3 and abscisic acid in relation to starch content and α-amylase activity

Somatic embryo quality is an important factor decisive for the efficiency of somatic embryogenesis. Addition gibberellic acid (GA₃) at a concentration of 1 μM to germination medium improved the regeneration of alfalfa somatic embryos. Inhibitory effect of ancymidol, an inhibitor of gibberellin biosynthesis, on germination and conversion may indicate that those processes require endogenous GAs. Since fluridone, an ABA biosynthetic inhibitor, at a concentration of 1 μM, had a slight stimulatory effect on germination of somatic embryos, it may be presumed that embryos contain a too high level of residual ABA after maturation phase (20 μM ABA is used at that phase). The observed improvement of regeneration of somatic embryos by GA₃ was correlated with acceleration of starch hydrolysis through α-amylase activity enhancement by GA₃. In contrast, the inhibitory effect of ABA on the above processes was probably related to inhibition of α-amylase activity and, in consequence, to delayed starch hydrolysis. It is suggested that α-amylase activity can be considered a good marker of the quality of Medicago sativa L. somatic embryos.     

The effects of low concentrations of aluminium on the growth and uptake of nitrate-N by white clover

Summary The effects of aluminium (Al³⁺) at concentrations of 0, 25, 50 and 100 μM on the growth of white clover, dependent upon N supplied as NO ₃ ⁻ , were examined in flowing solution culture. Plants were established with a normal nutrient supply for 7 weeks and then grown with carefully controlled pH (at 4.5) and P concentrations, and with 0, 25, 50 or 100 μM Al³⁺ for a further three weeks. There were rapid visual effects (i.e. symptoms of P deficiency and reduction in root extension) and the dry weights of shoots and roots were reduced at 50 and 100 μM. Less than 10% of Al absorbed from solution was transported to the shoots. The uptake of P, and its transport between roots and shoots, were reduced in plants grown with Al. The uptake of NO ₃ ⁻ stopped immediately after the introduction of 50 or 100 μM Al, and was significantly reduced at 25 μM after three weeks. During a second phase of the experiment, plants previously grown at 0, 25, 50 and 100 μM Al, were grown for a further 2 weeks either with NO ₃ ⁻ (with and without 50 μM Al³⁺) or without NO ₃ ⁻ but with inoculation by Rhizobia (and with or without 50 μM Al³⁺). The effects of the previous treatments with Al on N uptake were small during the second phase, but uptake by all plants was restricted when Al was present. Inoculation did not result in nodulation in the second phase when Al³⁺ was present in the solution, but Al already in the plant from the first phase did not prevent nodulation in the absence of Al during the second phase.     

Gibberellins in developing fruits of Pisum sativum cv. Alaska: Studies on their role in pod growth and seed development

Gibberellins A₁, A₈, A₂₀ and A₂₉ were identified by capillary gas chromatography-mass spectrometry in the pods and seeds from 5-d-old pollinated ovaries of pea (Pisum sativum cv. Alaska). These gibberellins were also identified in 4-d-old non-developing, parthenocarpic and pollinated ovaries. The level of gibberellin A₁ within these ovary types was correlated with pod size. Gibberellin A₁, applied to emasculated ovaries cultured in vitro, was three to five times more active than gibberellin A₂₀. Using pollinated ovary explants cultured in vitro, the effects of inhibitors of gibberellin biosynthesis on pod growth and seed development were examined. The inhibitors retarded pod growth during the first 7 d after anthesis, and this inhibition was reversed by simultaneous application of gibberellin A₃. In contrast, the inhibitors, when supplied to 4-d-old pollinated ovaries for 16 d, had little effect on seed fresh weight although they reduced the levels of endogenous gibberellins A₂₀ and A₂₉ in the enlarging seeds to almost zero. Paclobutrazol, which was one of the inhibitors used, is xylem-mobile and it efficiently reduced the level of seed gibberellins without being taken up into the seed. In intact fruits the pod may therefore be a source of precursors for gibberellin biosynthesis in the seed. Overall, the results indicate that gibberellin A₁, present in parthenocarpic and pollinated fruits early in development, regulates pod growth. In contrast the high levels of gibberellins A₂₀ and A₂₉, which accumulate during seed enlargement, appear to be unnecessary for normal seed development or for subsequent germination.Abbreviations GA_(a) gibberellin A_n - GC-MS combined gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - PFK perfluorokerosene - PVP polyvinylpyrrolidone