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PubMed2005

Modulation of nitrate reductase activity in rice seedlings under aluminium toxicity and water stress: role of osmolytes as enzyme protectant.

Nitrate reductase (NR) activity in the presence of Mg2+ (NR act) representing the non-phosphorylated NR state and the activity in the presence of EDTA (NR max) representing maximum NR activity was measured in roots and shoots of 15 d grown aluminium and water stressed rice seedlings to examine changes in NR activation state due to these stresses. Seedlings subjected to a moderate water stress level of -0.5 MPa for 24 h or grown in presence of 80 microM Al3+showed decreased level of NR max but resulted in higher NR act and NR activation state. However, seedlings grown in presence of a higher level of 160 microM Al3+ showed a decline in NR act as well as NR max. With a higher water stress Level of -2.0MPa a marked decline in the levels of both NR act and NR max was observed, whereas NR activation state remained almost unaltered with severe water stress. NR activity appeared to be sensitive to H2O2, PEG-6000, NaCl and various metal salts. Incorporation of these components in the enzyme assay medium led to decreased affinity of enzyme towards its substrate with increase in Km and decrease in Vmax values. Addition of each of the osmolytes i.e. 1 mol/L proline, 1 mol/L glycine betaine or 1 mol/L sucrose in the enzyme assay medium caused a considerable protection to the enzyme against the damaging effects of stressful components. An enhanced level of proline and glycine betaine was observed in Al-stressed seedlings and sucrose in Al as well as water stressed seedlings. Results suggest that Al toxicity and water stress decrease total amount of functional NR in rice seedlings and the osmolytes proline, glycine betaine and sucrose appear to have a direct protective action on enzyme NR under stressful conditions

Aluminum↗
PubMed2005

Concentration and release level of momilactone B in the seedlings of eight rice cultivars.

The release levels of a growth inhibitor, momilactone B, from rice (Oryza sativa L.) seedlings of eight cultivars were compared with the endogenous concentrations of momilactone B in their seedlings. All rice cultivars contained momilactone B in the seedlings, and their concentrations differed between the cultivars. Momilactone B was also found in all culture solutions in which these rice seedlings were grown, and the concentrations differed between the cultivars. The momilactone B concentrations in the culture solutions were reflected in the momilactone B concentrations in the seedlings. These results suggest that all rice cultivars may produce momilactone B and release momilactone B into the culture solutions. In addition, the release level of momilactone B may depend on the production level of momilactone B in the seedlings, which may affect allelopathic potential of these rice cultivars because as a growth inhibitor, momilactone B is able to act as an allelochemical.

Diterpenes↗
PubMed2005

Reactive oxygen metabolism in mycorrhizal and non-mycorrhizal citrus (Poncirus trifoliata) seedlings subjected to water stress.

The effect of the arbuscular mycorrhizal (AM) fungus, Glomus versiforme, on growth and reactive oxygen metabolism of trifoliate orange (Poncirus trifoliata) seedlings was studied in potted plants under well-watered (WW) and water stressed (WS) conditions. Water stress significantly decreased root colonization. Shoot dry weight, plant height and stem diameter were higher in AM than in non-AM seedlings regardless of the water status. Inoculation with G. versiforme increased root dry weight and leaf number per plant of WW seedlings. There was less malondialdehyde (MDA) concentration in leaves and roots of AM seedlings, as well as lower hydrogen peroxide (H(2)O(2)) and superoxide anion radical (O(2)(-)) concentrations in AM roots under WW and WS conditions. AM inoculation did not affect the H(2)O(2) and O(2)(-) concentrations of WW leaves. Whether WS or not, AM symbiosis notably increased the guaiacol peroxidase (G-POD) activity of leaves, glutathione reductase (GR) activity of leaves and ascorbate peroxidase (APX) activity of roots. AM infection also markedly increased the APX activity of WS leaves. Soluble proteins and glutathione (GSH) in leaves and roots and ascorbate (ASC) in leaves were higher in WW AM than in WW non-AM seedlings. AM infection also enhanced the ASC and GSH contents of leaves and roots in WS seedlings. Cross-tolerance might occur in AM plants and be enhanced by AM symbiosis. Our results suggest that the increased concentrations of antioxidant enzymes and non-enzymatic antioxidants found in AM plants may serve to protect the organism against oxidative damage, enhancing drought tolerance.

Antioxidants↗
PubMed2006

Pretreatment of seed with H2O2 improves salt tolerance of wheat seedlings by alleviation of oxidative damage and expression of stress proteins.

Increased salinity is a stringent problem to crop production while seed pretreatment can effectively induce salt tolerance in plants. Hydrogen peroxide (H(2)O(2)), a stress signal molecule, was evaluated as seed treatment to produce the metabolic changes, which could lead to improved salt tolerance in wheat. Soaking in 1, 40, 80 and 120 microM H(2)O(2) revealed a low penetration, reaching maximum at 5h (2.58+/-0.23 micro mol g(-1) fresh seeds at 120 microM) and declining thereafter to the level of water control by 8h. This revealed the activation of antioxidants and H(2)O(2) scavenging in seed after 5h. Seeds treated with 1-120 microM H(2)O(2) for 8h and germinated in saline (150 mM NaCl) medium curtailed the mean germination time (MGT) being even less than water controls. Level of H(2)O(2) in seedlings arising from H(2)O(2)-treated seeds grown under salinity was markedly lower than salinized controls, suggesting the operation of antioxidant system in them. These seedlings exhibited better photosynthetic capacity, particularly the stomatal conductance (gs), thus improving the leaf gas exchange due to stomatal component of photosynthesis. Moreover, H(2)O(2) treatment improved leaf water relations and maintained turgor. Although Na(+) and Cl(-) content increased due to salinity, H(2)O(2)-treated seedlings displayed greater tissue K(+), Ca(2+), NO(3)(-) PO(4)(3-) levels and improved K(+):Na(+) ratio. H(2)O(2) treatment enhanced the membrane properties, as revealed from greatly reduced relative membrane permeability (RMP) and less altered ion leakage pattern (comparable to water controls). Seedlings exhibited the expression of two heat-stable (stress) proteins with apparent molecular masses of 32 and 52 kDa. Results suggest that H(2)O(2) signals the activation of antioxidants in seed, which persists in the seedlings to offset the ion-induced oxidative damage. These changes led to the expression of stress proteins and improved physiological attributes, which supported the seedling growth under salinity.

Adaptation, Physiological↗
PubMed2006

Photoprotection, photosynthesis and growth of tropical tree seedlings under near-ambient and strongly reduced solar ultraviolet-B radiation.

Seedlings of two late-successional tropical rainforest tree species, Tetragastris panamensis (Engler) O. Kuntze and Calophyllum longifolium (Willd.), were field grown for 3-4 months at an open site near Panama City (9 degrees N), Panama, under plastic films that either transmitted or excluded most solar UV-B radiation. Experiments were designed to test whether leaves developing under bright sunlight with strongly reduced UV-B are capable of acclimating to near-ambient UV-B conditions. Leaves of T. panamensis that developed under near-ambient UV-B contained higher amounts of UV-absorbing substances than leaves of seedlings grown under reduced UV-B. Photosynthetic pigment composition, content of alpha-tocopherol, CO(2) assimilation, potential photosystem II (PSII) efficiency (evaluated by F(v)/F(m) ratios) and growth of T. panamensis and C. longifolium did not differ between seedlings developed under near-ambient and reduced solar UV-B. When seedlings were transferred from the reduced UV-B treatment to the near-ambient UV-B treatment, a pronounced inhibition of photosynthetic capacity was observed initially in both species. UV-B-mediated inhibition of photosynthetic capacity nearly fully recovered within 1 week of the transfer in C. longifolium, whereas in T. panamensis an about 35% reduced capacity of CO(2) uptake was maintained. A marked increase in UV-absorbing substances was observed in foliage of transferred T. panamensis seedlings. Both species exhibited enhanced mid-day photoinhibition of PSII immediately after being transferred from the reduced UV-B to the near-ambient UV-B treatment. This effect was fully reversible within 1d in T. panamensis and within a few days in C. longifolium. The data show that leaves of these tropical tree seedlings, when developing in full-spectrum sunlight, are effectively protected against high solar UV-B radiation. In contrast, leaves developing under conditions of low UV-B lacked sufficient UV protection. They experienced a decline in photosynthetic competence when suddenly exposed to near-ambient UV-B levels, but exhibited pronounced acclimative responses.

Biomass↗
PubMed2004

Light-responsive subtilisin-related protease in soybean seedling leaves.

Protease C1 (E.C. 3.4.21.25), the soybean (Glycine max L. Merrill) proteolytic enzyme responsible for initiating the degradation of soybean storage proteins in seedling cotyledons appears at even higher levels in seedling leaves. This was manifested at the mRNA level through northern blot analysis, at the protein level through western blot analysis, through determination of enzyme activity, and also through isolation and partial sequencing of active leaf enzyme. Comparison of cDNA and amino acid sequences, as well as characterization of enzyme activity, is consistent with the leaf enzyme being identical to or highly similar to the cotyledon enzyme. Protease C1 mRNA and protein are also present in stems of soybean seedlings, but is very low to absent in the roots. This presence in the aerial tissues is consistent with the higher steady state level of gene expression at both the mRNA and protein levels when the seedlings are grown in a 12-h light: 12-h dark photoperiod as compared to seedlings grown in continuous darkness. Transfer of dark-grown seedlings to light is followed by marked elevation in protease C1 protein as seen in western blots.

Amino Acid Sequence↗
PubMed2004

Effects of adding organic materials to an acid sulfate soil on the growth of cocoa (Theobroma cacao L.) seedlings.

Acid sulfate soils having a pH of less than 3.5 are widespread in Malaysia. Some of these soils are planted to cocoa, but the yield is reported to be low due to soil infertility related to Al toxicity. Cocoa growth is sensitive to the presence of Al in the soil. To a certain extent, Al toxicity in soils can be reduced by organic matter application and to a greater extent in iron-poor acid sulfate soil. A study was conducted to determine the efficacy of various types of organic materials easily available in the country to ameliorate acid sulfate soil infertility for growing cocoa seedlings. The treatments were control (nil), lime (3 t/ha), peat (10% w/w), peat plus green manure (10% w/w), peat plus rice straw (10% w/w), peat plus chicken dung (10% w/w) and peat plus POMS (Palm oil mill sludge) (10% w/w). The growth of cocoa seedlings was affected significantly by the presence of Al in the cocoa tissues. As the amount of Al in the leaves increased, the relative top dry weight of cocoa seedlings decreased. Likewise, the relative plant height was negatively correlated with Al in the leaves. Peat as well as peat in combination with green manure, rice straw, chicken dung or palm oil mill sludge was able to reduce Al toxicity in acid sulfate soil; the highest top dry weight of cocoa seedlings were obtained in the peat plus green manure treatment. The best cocoa seedlings root growth was found for the peat treatment alone. The relative top dry weight of cocoa seedlings was negatively correlated with Al(3+) as well as Al(3+)+Al(OH)(2+)+Al(OH)(2)(+) activity in the soil solution. The critical values for Al(3+) and the combination of Al(3+)+Al(OH)(2+)+Al(OH)(2)(+) activity in the soil solution were 10 microM and 15 microM, respectively.

Aluminum↗
PubMed2004

Release and activity of allelochemicals from allelopathic rice seedlings.

3-Isopropyl-5-acetoxycyclohexene-2-one-1 (1), momilactone B (2), and 5,7,4'-trihydroxy-3',5'-dimethoxyflavone (3) were isolated and identified from an allelopathic rice accession PI312777. These three compounds at low concentrations could inhibit the growth of weeds Echinochloa crusgalli and Cyperus difformis associated with rice, especially mixtures of the compounds had stronger inhibitory activity than did individual compounds. Studies with hydroponic culture, continuous root exudates trapping system (CRETS), and direct resin adsorption methods showed that a total of 7.6 n moles 1, 2, and 3 were exuded from living roots of each seedling into the environment at 10 days after seedlings were transplanted. Furthermore, 1, 2, and 3 were found in the soil growing PI312777 seedlings at day 15 after seedlings emergence and reached a total of 39.5 microg/g soil at day 30. The results indicated that PI 312777 seedlings could release sufficient quantities of 1, 2, and 3 into the environment to act as allelochemicals inhibiting the growth of associated weeds. Investigations on the distribution of 1, 2, and 3 in PI 312777 plant, and its root exudates showed that the levels of 1, 2, and 3 were significantly higher in the shoots and root exudates than in the roots, and only trace 1 was observed in the roots. The results suggest that the roots of rice seedlings are not major site of synthesis or accumulation 1, 2, and 3, but a pathway for their release into the environment. The levels of 1, 2, and 3 in the root exudates were over 2-folds higher under direct resin adsorption than under hydroponic culture and CRETS, and hence, it is the preferred method to collect and identify active allelochemicals in rice exudates in future studies on rice allelopathy.

Herbicides↗
PubMed2003

Drought stress alters the concentration of wood terpenoids in Scots pine and Norway spruce seedlings.

Drought is known to have an impact on the resistance of conifers to various pests, for example, by affecting resin flow in trees. Little is known, however, about the quantitative and qualitative changes in resin when trees are growing in low moisture conditions. We exposed Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings to medium and severe drought stress for two growing seasons and analyzed the monoterpenes and resin acids in the main stem wood after two years of treatment. In addition to secondary chemistry, we measured the level of nutrients in the needles and the growth response of seedlings. After the first year of treatment, drought stress did not affect the growth of seedlings, but in the second year, shoot growth was retarded, especially in Scots pine. In both conifer species, severe drought increased the concentrations of several individual monoterpenes and resin acids. Total monoterpenes and resin acids were 39 and 32% higher in severe drought-treated Scots pine seedlings than in the controls, and 35 and 45% higher in Norway spruce seedlings. In Scots pine needles, the concentrations of nitrogen and phosphorus increased, while magnesium and calcium decreased compared to controls. In Norway spruce needles, nutrient concentrations were not affected. The results suggest that drought stress substantially affects both the growth of conifers and the chemical quality of the wood. We discuss the potential trade-off in growth and defense of small conifer seedlings.

Adaptation, Physiological↗
PubMed2003

Accumulation of LEA proteins in salt (NaCl) stressed young seedlings of rice (Oryza sativa L.) cultivar Bura Rata and their degradation during recovery from salinity stress.

Germination and subsequent hydroponic growth under salt stress (100 mmol/L NaCl) triggered an accumulation of six major stress proteins and resulted in a growth arrest of young seedlings of rice (Oryza sativa L.) cv. Bura Rata. Based on two-dimensional electrophoretic resolution, partial amino acid sequencing and immunodetection techniques, four of the salt stress-induced polypeptides were identified as LEA proteins. Under all experimental conditions wherein seedlings exhibited superior halotolerance, salt stress-induced LEA proteins were expressed at low levels. In contrast, accumulation of LEA proteins was found associated with growth arrest. When returned to non-saline media, seedlings stressed with salt for four days recovered immediately. Longer exposure to 100 mmol/L NaCl, however, progressively delayed recovery and reduced the number of seedlings which could recover from salt stress. Recovery from salt stress was consistently accompanied by degradation of the salt stress-induced LEA proteins. The results of this study show that LEA proteins accumulate during the salinity-triggered growth arrest of young Bura Rata seedlings and are mobilised during the recovery of seedlings from salinity stress.

Amino Acid Sequence↗
PubMed2004

Physiological and ecological significance of sunflecks for dipterocarp seedlings.

Irradiance is highly dynamic in many plant canopies. Photosynthesis during sunflecks provides 10-90% of daily carbon gain. The survivorship of tree seedlings in the deeply shaded understorey of tropical rain forests is limited by their ability to maintain a positive carbon balance. Dipterocarp seedlings from the SE Asian rain forest were used as a model system to test novel aspects of the physiological and ecological significance of sunflecks. First, understorey seedlings experienced leaf temperatures up to 38 degrees C in association with sunflecks. Under controlled environment conditions, the inhibition of carbon gain at 38 degrees C, compared with 28 degrees C, was significantly greater during a sequence of sunflecks (-59%), than under uniform irradiance (-40%), providing the same total photosynthetic photon flux density (PPFD). Second, the relative enhancement effects of elevated [CO2] were greater under sunflecks (growth +60%, carbon gain +89%), compared with uniform irradiance (growth +25%, carbon gain +59%), supplying the same daily PPFD. Third, seedling growth rates in the forest understorey were 4-fold greater under a dynamic irradiance treatment characterized by long flecks, compared with a regime of short flecks. Therefore, stresses associated with dynamic irradiance may constrain photosynthetic carbon gain. Additionally, seedling photosynthesis and growth may be more responsive to interactions with abiotic factors, including future changes in climate, than previously estimated. The sensitivity of seedling growth to varying patterns of dynamic irradiance, and the increased likelihood of species-specific responses through interactions with environmental factors, indicates the potential for sunflecks to influence regeneration processes, and hence forest structure and composition.

Acclimatization↗
PubMed2002

Responses of transpiration and photosynthesis to reversible changes in photosynthetic foliage area in western red cedar (Thuja plicata) seedlings.

Experiments were conducted on 1-year-old western red cedar (Thuja plicata Donn.) seedlings to determine the response of illuminated foliage to reversible changes in total photosynthetic foliage area (L(A)). Reductions in L(A) were brought about by either shading the lower foliage or by reducing the ambient CO2 concentration (c(a)) of the air surrounding the lower part of the seedling. In the latter case, the vapor pressure was also changed so that transpiration rates (E) could be manipulated independently of photosynthetic rates (A). We hypothesized that following such treatments, short-term compensatory changes would occur in stomatal conductance (g(s)) and A of the remaining foliage. These changes would occur in response to hydraulic signals generated by changes in the water potential gradient rather than changes in the distribution of sources and sinks of carbon within the seedling. When a portion of the foliage was shaded, there was an immediate reduction in whole-seedling E and a concomitant increase in g(s), A and E in the remaining illuminated foliage. However, the intercellular CO2 concentration did not change. These compensatory effects were fully reversed after the shade was removed. When the lower foliage A was reduced to < 0 micromol m-2 s-1, by shading or lowering c(a), and E was either unchanged or increased (by adjusting the vapor pressure deficit), there was no significant increase in g(s) and A in the remaining foliage. We conclude that compensatory responses in illuminated foliage occur only when reductions in L(A) are accompanied by a reduction in whole-plant E. The relationship between the reduction in whole-seedling E and the increase in A is highly linear (r2 = 0.68) and confirms our hypothesis of the strong regulation of g(s) by hydraulic signals generated within the seedling. We suggest that the mechanism of the compensatory effects is a combination of both increased CO2 supply, resulting from increased g(s), and a response of the rate of carboxylation, possibly related to the activity of Rubisco.

Carbon Dioxide↗
PubMed2004

Growth, allocation and tissue chemistry of Picea abies seedlings affected by nutrient supply during the second growing season.

One-year-old Norway spruce (Picea abies (L.) Karst.) seedlings were grown hydroponically in a growth chamber to investigate the effects of low and high nutrient availability (LN; 0.25 mM N and HN; 2.50 mM N) on growth, biomass allocation and chemical composition of needles, stem and roots during the second growing season. Climatic conditions in the growth chamber simulated the mean growing season from May to early October in Flakaliden, northern Sweden. In the latter half of the growing season, biomass allocation changed in response to nutrient availability: increased root growth and decreased shoot growth led to higher root/shoot ratios in LN seedlings than in HN seedlings. At high nutrient availability, total biomass, especially stem biomass, increased, as did total nonstructural carbohydrate and nitrogen contents per seedling. Responses of stem chemistry to nutrient addition differed from those of adult trees of the same provenance. In HN seedlings, concentrations of alpha-cellulose, hemicellulose and lignin decreased in the secondary xylem. Our results illustrate the significance of retranslocation of stored nutrients to support new growth early in the season when root growth and nutrient uptake are still low. We conclude that nutrient availability alters allocation patterns, thereby influencing the success of 2-year-old Norway spruce seedlings at forest planting sites.

Biomass↗
PubMed2004

Water stress decreases the transfer conductance of Douglas-fir (Pseudotsuga menziesii) seedlings.

We tested the hypothesis that transfer conductance (gi) of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings is reduced by water stress. Seedlings were irrigated with a solution of 25% polyethylene glycol so as to impose water stress rapidly, thereby limiting acclimatory responses. Transfer conductance was measured pre-treatment and post-treatment by two methods. Water stress reduced net photosynthesis by 20-50%. The initial slope of the rate of photosynthesis (A) over the intercellular carbon dioxide (CO2) concentration (Ci) response was reduced by water stress, indicating that reduced photosynthesis was not wholly accounted for by reduced stomatal conductance. The carbon isotope and chlorophyll fluorescence methods both indicated that water stress decreased gi. From isotopic measurements with 1% O2, gi was 0.076 +/- 0.009 (mean +/- SE) mol m(-2) s(-1) in well-watered seedlings and 0.044 +/- 0.004 mol m(-2) s(-1) in water-stressed seedlings. Fluorescence estimates of gi were 0.08 +/- 0.01 mol m(-2) s(-1) in well-watered seedlings and 0.044 +/- 0.004 mol m(-2) s(-1) in water-stressed seedlings. The drought-induced reduction in gi was responsible for the reduction in slope of the A/Ci response, and thus there was no difference in the slope of the A over the chloroplastic CO2 concentration (Cc) response between treatments and no indication of impaired mesophyll metabolism. These data illustrate that impairments of mesophyll metabolism can be revealed only from analysis of the A/Cc response.

Dehydration↗
PubMed2005

Effects of soil temperature on shoot and root growth and nutrient uptake of 5-year-old Norway spruce seedlings.

Soil temperature is a main factor limiting root growth in the boreal forest. To simulate the possible soil-warming effect of future climate change, 5-year-old Norway spruce (Picea abies (L.) Karst.) seedlings were subjected to three simulated growing seasons in controlled environment rooms. The seedlings were acclimated to a soil temperature of 16 degrees C during the first (GS I) and third growing seasons (GS III), but were assigned to random soil-temperature treatments of 9, 13, 18 and 21 degrees C during the second growing season (GS II). In GS II, shoot diameter growth was lowest in the 21 degrees C treatment and root growth was lowest in the 9 degrees C treatment. In GS III, shoot height and root length growth improved in seedlings that had been kept at 9 degrees C during GS II, indicating compensatory growth in response to increased soil temperature. The temporary decrease in soil temperature had no long-lasting significant effect on seedling biomass or total nutrient uptake. At the end of GS III, fine roots of seedlings exposed to a soil temperature of 21 degrees C in GS II were distributed more evenly between the organic and mineral soil layers than roots of seedlings in the other treatments. During GS II and GS III, root growth started earlier than shoot growth, decreased during the rapid shoot elongation phase and increased again as shoot growth decreased.

Picea↗
PubMed2006

Spacing of silver birch seedlings grown in containers of equal size affects their morphology and its variability.

Silver birch (Betula pendula Roth) seedlings were grown individually in containers arranged in rows radiating from a central point (Nelder plot) at densities spanning the range from 207 to 891 plants m(-2). Height of one set of seedlings was measured at weekly intervals and additional seedlings were harvested each week for dry mass and leaf area measurements. Height and shoot:root dry mass ratio increased with plant density. Seedling-to-seedling variability in dry mass, but not height, increased with increasing plant density. The red to far-red (R:FR) photon ratio in horizontally propagated radiation decreased with increasing density, even when plant densities and leaf area index values were low. In a separate experiment, elongating internodes of seedlings were irradiated locally by red and far-red light emitting diodes and stem elongation measured. Far-red light markedly increased stem elongation, suggesting that changes in light quality sensed by growing internodes are involved in the observed responses to growth density.

Betula↗
PubMed2006

Cotyledon damage at the seedling stage affects growth and flowering potential in mature plants.

Seedling herbivory is an important selective filter influencing patterns of plant community composition. Nevertheless, while many of the mechanisms governing seedling selection by herbivores are well established, the effects of tissue loss at the seedling stage on subsequent plant development are poorly understood. Here we examined how the removal of 50 or approximately 100% of cotyledon area from 7-d-old chalk grassland seedlings affected subsequent plant growth and flowering over a 100-d period. Cotyledon damage had a significant effect on growth during the establishment phase for six of the nine species. For two species, significant effects on plant growth were manifest in 100-d-old plants. Of the five species that flowered, three developed fewer inflorescences or flowered later as a consequence of cotyledon damage suffered as a seedling. Our results show that, in addition to the direct effect of herbivory on seedling mortality, more subtle sublethal effects may also influence plant establishment. Reduced growth as a result of cotyledon damage may have implications for plant competition during the establishment phase, and on subsequent reproductive success at maturity.

Asteraceae↗
PubMed2005

Similarities and differences between the characteristics of gibberellin-binding protein and gibberellin 2-oxidases in adzuki bean (Vigna angularis) seedlings.

Gibberellin-binding proteins (GBPs) were purified ca. 230,000 fold. The characteristics of adzuki GBP were examined and compared with those of a recombinant gibberellin 2-oxidase (rVaGA2oxA1) that was fused with glutathione S-transferase (GST). VaGA2oxA1 was most abundantly expressed in etiolated adzuki bean seedlings, and VaGA2oxA1 and GBPs from adzuki bean seedlings showed gibberellin-binding activity when incubated with 2-oxoglutarate and Co2+. Both rVaGA2oxA1 and partially purified GBPs from adzuki bean seedlings showed very similar selectivity to gibberellins in binding assays, where biologically active gibberellins such as GA4, GA3, GA7, and GA1 showed higher binding affinity than biologically inactive gibberellins such as GA8, GA34, and 3-epi-GA4. The polyclonal antibody raised against rVaGA2oxA1 cross-reacted with all rVaGA2oxs (rVaGA2oxA1, rVaGA2oxA2, rVaGA2oxB1, rVaGA2oxB2, and rVaGA2oxB3) whose cDNAs were cloned from adzuki bean seedlings. Treated with the antibody, the recombinants that originally showed gibberellin-binding activity lost both binding activity and enzymatic activity. In contrast to the recombinants, the gibberellin-binding activity of GBPs from adzuki bean seedlings was hardly affected by the antibody treatment. The GBPs showed very weak gibberellin 2-oxidase-like activity, and it was not affected by the antibody treatment either. These observations suggest that a major component that showed GA-binding activity was apparently different from any gibberellin 2-oxidase cloned from the seedlings.

Carrier Proteins↗