Carbohydrate Status of Tulip Bulbs during Cold-Induced Flower Stalk Elongation and Flowering

The effect of a cold treatment on the carbohydrate status of the scales and flower stalk of Tulipa gesneriana L. cv Apeldoorn bulbs during growth after planting was studied and compared with bulbs not given cold treatment. Bulbs were stored dry for 12 weeks at 5[deg]C (precooled) or 17[deg]C (noncooled). Only the 5[deg]C treatment led to rapid flower stalk elongation and flowering following planting at higher temperatures. Precooling enhanced mobilization of starch, fructans, and sucrose in the scales. The cold-stimulated starch breakdown was initially accompanied by increased [alpha]-amylase activity per scale. In noncooled bulbs, [alpha]-amylase activity slightly decreased or remained more or less constant. Cold-induced flower stalk elongation was partially accompanied by a decrease in the sucrose content and an increase in the glucose content and invertase activity per g dry weight. The starch content in internodes initially decreased and subsequently increased; [alpha]-amylase activity per g dry weight of the lowermost internode showed a peak pattern during starch breakdown and increased thereafter. The internodes of noncooled bulbs, on the contrary, accumulated sucrose. Their glucose content and invertase activity per g dry weight remained low. Starch breakdown was not found and [alpha]-amylase activity per g dry weight of the lowermost internode remained at a low level. Precooling of tulip bulbs thus favors reserve mobilization in the scales and flower stalk and glucose accumulation in the elongating internodes.     

Carbon allocation in developing spruce needles. Enzymes and intermediates of sucrose metabolism

In lyophilized needles of Norway spruce ( Picea abies [L.] Karsten) and starting from bud break, we determined enzyme activities (sucrose phosphate synthase [SPS; EC 2.4,1.14]. sucrose synthase [SS; EC 2.4,1.13]. acid invertase [AI; EC 3.2,1.26]) and intermediates (starch, sucrose, glucose, fructose; fructose 6-phosphate, fructose 2.6-bisphosphate [F26BP]) of carbohydrate metabolism together with needle weight, shoot length, chlorophyll and protein. For up to 110 days after bud break, samples were taken twice a week from about 25-year-old trees under field conditions. At least three periods can be distinguished during needle maturation. During the first period (up to 45 days after bud break) Al showed the highest extractable activity. This coincided with very high levels of F26BP (up to 11 pmol [mg dry weight]⁻¹) and a transient increase of starch in parallel to a decrease of sucrose. The interval between 45 and 70 days after bud break was characterized by high SS activity (ratio of fructose/glucose >1), much decreased levels of F26BP (down to below 1 pmol [mg dry weight]⁻¹), and a pronounced increase in the dry weight/fresh weight ratio. In parallel, starch declined and soluble carbohydrates increased. Finally, needle maturation was characterized by decreasing SS and continuously increasing SPS activities, so that the ratio of SPS/SS increased more than 6-fold. AI. however, did not decline with maturation. Changes in pool sizes of metabolites and enzyme activities (AI. SPS) are consistent with current concepts on sink/source transition. SS is obviously important with regard to the synthesis of structural polysaccharides.     

Invertase and sucrose synthase activity, carbohydrate status and endogenous IAA levels during Citrus leaf development

Levels of activity of the sucrose catabolizing enzymes, acid invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13), were measured during development of new leaves of Citrus sinensis (L.) Osbeck cv. Shamouti. Soluble acid invertase showed a peak activity of 32 nkat (g fresh weight)⁻¹ at ca 60% of full leaf expansion and rapidly declined toward and after full expansion. There was no concomitant increase in an insoluble form of the enzyme. Sucrose synthase activity, measured in the synthesis direction, declined from 33% of full leaf expansion [10 nkat (g fresh weight)⁻¹] 10, and following, full expansion. Highest sucrose synthase activity, measured in the cleavage direction, was 6 nkat (g fresh weight)⁻¹ and showed little change during development. Acid invertase has a K_m of 5 m M for sucrose, while sucrose synthase had a K_m of 118 m M for sucrose. Changes in acid invertase activity correlated with changes in the reducing sugar:sucrose ratio. These results suggest that soluble acid invertase activity is the primary enzyme responsible for sucrose catabolism in the expanding Citrus leaf. Changes in leaf expansion rate and invertase activity did not correlate positively with changes in endogenous free IAA level, as determined by enzyme linked immunoassay.     

Content of low-molecular-weight thiols during the imbibition of Pea seeds

The metabolism of low-molecular-weight thiols was investigated in seeds of Pisum sativum L. cv. Kleine Rheinländerin during imbibition in water for 14 h. The amount of oxidized glutathione (GSSG) decreased from 319 nmol (g dry weight)⁻¹ in dry seeds to 38 nmol (g dry weight)⁻¹ within the first 14 h of imbibition. The decrease may have been due to the reduction of GSSG to reduced glutathione (GSH), catalyzed by the enzyme glutathione reductase (GR; EC 1.6.4.2). The enzyme activity was high in dry seeds [25 nkat (g dry weight)⁻¹] and decreased to 20 nkat (g dry weight)⁻¹ within 14 h of imbibition. The activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) decreased from 100 nkat (g dry weight)⁻¹ in dry seeds to 67 nkat (g dry weight)⁻¹ after 14 h of imbibition. Within 14 h the amount of γ-glutamyl-cysteine (γ-GC) decreased from 135 to 38 nmol (g dry weight)⁻¹, whereas the cysteine content rose from 81 nmol (g dry weight)⁻¹ in dry seeds to a maximum of 170 nmol (g dry weight)⁻¹ after 12 h of imbibition, which may be due to the degradation of γ-GC into cysteine.     

Metabolic Changes Associated with the Sink-Source Transition During Sprouting of the Axillary Buds on the Sugarcane Culm

Sucrose, glucose and fructose concentrations, and sucrolytic enzyme activities were measured in the developing shoots and internodes of sprouting sugarcane setts (Saccharum spp, variety N19). The most striking change during the sink-source transition of the internode and germination of the axillary bud is a more than five-fold induction of cell wall invertase in the germinating bud. In contrast, soluble acid invertase is the main sucrose hydrolytic activity induced in the internodal tissue. A cycle of breakdown and synthesis of sucrose was evident in both the internodes and the shoots. During shoot establishment, the sucrose content decreased and the hexose content increased in the internodal tissues while both sucrose and hexoses continuously accumulated in the shoots. Over the sprouting period internode, dry mass was reduced by 25 and 30 % in plants incubated in a dark/light cycle or total darkness, respectively. Sucrose accounted for 90 % of the dry mass loss. The most significant changes in SuSy activity are in the synthesis direction in the shoots resulting in a decrease in the breakdown/synthesis ratio. In contrast the SuSy activity in the internodal tissue decrease and more so in the synthesis activity resulting in an increase in the breakdown to synthesis ratio.     

Gibberellin-induced elongation and osmoregulation in internodes of floating rice

Internodal elongation in floating rice ( Oryza sativa L. cv. Habiganj Aman II) is known to be enhanced by treatment with ethylene or gibberellic acid (GA₃) at high relative humidity (RH). However, ethylene-induced internodal elongation is inhibited at low RH. while GA₃-induced internodal elongation is hardly affected by humidity. We examined the possible involvement of osmoregulation in the stimulation by GA₃ of the elongation of internodes at low RH. Submergence and treatment with ethylene or GA3₃ at 100% RH increased the osmotic potential in internodes of excised stem segments, while GA₃ at 20% RH maintained the osmotic potential at a low level. In internodes of stem segments that had been treated with GA₃ at 20% RH, the activity of invertase and the level of soluble sugars were almost 2- and 1.5-fold higher, respectively, than those in internodes that had been treated with GA₃ at 100% RH. These results indicate that one of the possible mechanisms by which GA₃ promotes elongation of internodes at low RH involves the osmoregulation that is achieved by promotion of the synthesis of invertase.     

Invertase Activity, Carbohydrate Metabolism and Cell Expansion in the Stem of Phaseolus vulgaris L.

In the stem of Phaseolus vulgaris L. the specific activity ofacid invertase was highest in the most rapidly elongating internode.Activity of the enzyme was very low in internodes which hadcompleted their elongation, in young internodes before the onsetof rapid elongation, and in the apical bud. From shortly afterits emergence from the apical bud the elongation of internode3 was attributable mainly to cell expansion. Total and specificactivities of acid invertase in this internode rose to a maximumat the time of most rapid elongation and then declined. Transferof plants to complete darkness, or treatment of plants withgibberellic acid (GA₃), increased the rate of internode elongationand final internode length by stimulating cell expansion. Bothtreatments rapidly increased the total and specific activitiesof acid invertase in the responding internodes; peak activitiesof the enzyme occurred at the time of most rapid cell expansion. In light-grown plants, including those treated with GA₃, rapidcell and internode elongation and high specific activities ofacid invertase were associated with high concentrations of hexosesugar and low concentrations of sucrose. As cell growth ratesand invertase activities declined, the concentration of hexosefell and that of sucrose rose. In plants transferred to darkness,stimulated cell elongation was accompanied by a rapid decreasein hexose concentration and the disappearance of sucrose, indicatingrapid utilization of hexose. No sucrose was detected in theapical tissues of light-grown plants. The results are discussed in relation to the role of acid invertasein the provision of carbon substrates for cell growth. Key words: Cell expansion, Acid invertase, Hexose, Sucrose, Phaseolus     

Long distance transport of potassium in cereals during grain filling in intact plants

Translocation of labeled potassium (K⁺) from the root to the ear and its distribution within the culm during 4, 8 and 12 h of uptake was studied in intact wheat plants ( Triticum aestivum L. cv. Kolibri) 3 and 5 weeks after anthesis at 0.5 and 5.0 m M K⁺ concentration in the uptake solution. Uptake of labeled K⁺ into the shoot was proportional to the K⁺ concentration applied. After 4 h of uptake about 2% and after 12 h about 7% of labeled K⁺ applied to the roots were taken up into the shoot at both K⁺ concentrations. After 12 h of uptake only 6% of the total label in the culm had reached the ear, while about 40% of the label was found in the upper three internodes. In spite of an increasing concentration of labeled K⁺ during 12 h in the uppermost internode (peduncle), translocation of K⁺ into the rachis was low. The low and uniform K⁺ content found generally in grain dry weight seems therefore to be due to a controlled K⁺ supply to the ear.     

The activity and interaction of brassinolide and gibberellic acid in mung bean epicotyls

The growth rate of mung bean epicotyls was used for evaluating the effect of brassinolide on cell elongation. Growth above that of control plants was observed at 10⁻¹⁰ M and above. Gibberellic acid showed an additivity relationship with low concentrations (10⁻⁹–10⁻⁸ M ) of brassinolide in this test system and the two growth promoters may therefore act independently at the cellular level. Because of relative ease of operation, great sensitivity and the short time required for assessing biological activity, this assay could be used in conjunction with the bean second internode bioassay for evaluating the activity of brassinolide and its analogs, as well as of other growth promotors.     

Photosynthesis and photoassimilation of glucose during photomixotrophic growth of Marchantia polymorpha cells in suspension culture

Even in the presence of glucose the growth of Marchantia polymorpha L. (cell line HYH-2F) requires light, and growth is more sensitive to 10⁻⁶ M 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea than to 10⁻⁴ Antimycin A. The inability of the cells to grow in the dark is due to the low level of respiration. The respiration rate under light increased to four times the dark value. The values of the compensation ratio (the photosyntehtic rate/the respiration rate) for the oxygen exchange were below 1.0 daring the growth period, although oxygen evolution was found. At the early exponential phase, oxygen evolution was 0.373 μmol (mg cell dry weight)⁻¹ h⁻¹ [61.7 μmol (mg chlorophyll)⁻¹ h⁻¹]. M. polymorpha cells are unable to grow anaerobically in the light without a supply of carbon dioxide. When 1% carbon dioxide in nitrogen is supplied, photochemically produced oxygen and energy are sufficient for sustained growth although at significantly reduced yields in both cell dry weight and chlorophyll. Photosyntehtic CO₂ assimilation rate was 0.13 μmol (mg cell dry weight)⁻¹ h⁻¹[11.3 μmol (mg chlorophyll)⁻¹ h⁻¹]. At least one-third of the carbon atoms in cellular constituents seem to be derived from atmospheric carbon dioxide, which indicates that M. polymorpha cells grow photomixotrophicaily.     

Oxygen uptake in developing eggs and larvae of the cod, Gadus morhua L.

Unfertilised cod eggs showed a mean oxygen uptake rate at 5°C of 0.089 μl O₂, dry wt.⁻¹ h⁻¹; this gradually rose to 0.768 μl O₂ mg dry wt.⁻¹ h⁻¹ in eggs about to hatch. From hatching to complete yolk absorption larvae respired at 1.6 μl O₂, mg dry wt.⁻¹ h⁻¹. During starvation following yolk absorption, uptake fell significantly to 1.1 μl O₂, mg dry ⁻¹ h⁻¹. Much of this decrease in oxygen consumption was shown to be caused by reduction in activity. Loss of weight during the embryo and larval phases could not easily be reconciled with total oxygen consumption; it is suggested that cod embryos and larvae may not rely solely upon endogenous energy reserves during development.     

Spiral growth and cell wall properties of the gibberellin-treated first internodes in the seedlings of a wheat cultivar tolerant to deep-sowing conditions

The Hong Mang Mai wheat cultivar is tolerant to deep-sowing conditions because it has an elongated first internode that is sensitive to gibberellin (GA₃). The cells in the GA-treated first internodes were approximately 4.2 mm long, twice as long as the untreated Hong Mang Mai first internode cells. The elongation of the first internode of Hong Mang Mai, particularly when treated with GA₃, was accompanied by remarkable spiral growth. In contrast, the first internodes of the GA-insensitive cultivar Norin 10 did not exhibit GA₃-induced elongation or spiral growth. The walls of the first internode cells of GA₃-treated Hong Mang Mai seedlings showed increased extensibility and higher (1→3), (1→4)- β - d -glucanase activity, autolysis and glucan contents than the cell walls of untreated Hong Mang Mai first internodes. The changes in the cell wall extensibility due to GA₃ treatment correlated strongly with the GA₃-induced changes in cell wall glucan content, autolysis, and glucanase activity. GA₃-treated Hong Mang Mai seedlings showed elevated expression of Glucanase EI gene in the first internode compared to GA₃-treated Norin 10. Thus, GA aids first internode elongation in Hong Mang Mai by enhancing glucan turnover and thus increasing cell wall loosening. The spiral growth of the first internode also helps the plant elongate against soil resistance, thereby promoting the deep-sowing tolerance of this cultivar.     

Influence of spectral quality on the growth response of intact bean plants to brassinosteroid, a growth-promoting steroidal lactone. I. Stem elongation and morphogenesis

The growth response of bean ( Phaseolus vulgaris L. 'Pinto') plants treated with 5 μg of brassinosteroid (BR) in the bean second-internode assay was measured in a controlled environment under 3 radiation sources: cool white fluorescent (CWF), far-red (FR) fluorescent and incandescent (INC) lamps. Growth comparisons were made under equal levels (90 μmol s^-1 m^-2) of photosynthetic photon flux density provided by CWF or INC lamps and equal levels of far-red (28 W m^-2, 700–800 nm) radiation provided by the same INC or FR lamps. Treatment of the second internode with BR produced a sequential increase in elongation, curvature, and swelling under normal bioassay conditions (CWF lamps), as observed previously with brassins. In addition, BR induced marked splitting of the treated internode provided that ample photosynthate was available. Spectral quality had a differential effect on internodal elongation. Under CWF lamps, internodes, 6 days after BR-treatment, were 2–3 times longer than those of controls; under INC or FR lamps they were 30–60% shorter than those of controls. In all cases, BR-treatment greatly stimulated accumulation of photosynthate in the treated internode, as indicated by fresh and dry weights and stem diameter measurements. This suggests a possible mobilization role for BR in the intact plant. Brassinosteroid also partially overcame the natural inhibitory effects of CWF radiation on stem elongation.     

Effects of exogenous gibberellins and paclobutrazol on floral stalk growth of tulip sprouts isolated from cooled and non-cooled tulip bulbs

The involvement of gibberellins (GAs) in the regulation of floral stalk elongation and flower development has been studied in tulip. The biological activity of GA₄ and GA₉, both endogenous in tulip bulb sprouts, and GA₁, was tested in vitro on sprouts of cooled and non-cooled tulip bulbs ( Tulipa gesneriana L. cv. Apeldoorn), in the presence or absence of the GA biosynthesis inhibitor paclobutrazol. At early starting dates of incubation, floral stalks from both cooled and non-cooled bulbs hardly showed any elongation in the absence of exogenous GA. Paclobutrazol had no effect on floral stalk elongation, and the response to GAs of sprouts from cooled bulbs was greater than that of sprouts from non-cooled bulbs. At later starts of incubation, considerable floral stalk elongation occurred without GA application. Paclobutrazol inhibited this floral stalk elongation, and the growth of sprouts from both cooled and non-cooled bulbs was stimulated by GA application. The effect of paclobutrazol was reversed by simultaneous application of GA₄ or GA₉. Application of GA with and without paclobutrazol resulted in the same elongation of the floral stalk, indicating the absence of substantial side effects of the inhibitor. The isolated sprouts did not develop a full-grown flower without the addition of GA. GA₄ was more effective than GA₉ in stimulating this flower development. The results demonstrate that both sprouts from cooled and non-cooled bulbs are responsive to exogenous GAs in vitro, and may be a site of GA biosynthesis.     

Sucrose‐cleaving enzymes and carbohydrate pools in Lilium longiflorum floral organs

The activities of soluble invertase (EC 3.2.1.26), cell wall invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13) were determined in Easter lily ( Lilium longiflorum Thunb. cv. Nellie White) floral organs during flower development. These enzyme activities were correlated with dry weight gains and carbohydrate pools to investigate the importance of their expression in maintaining sink strength of floral organs. In the early stages of flower bud development, anthers exhibited the highest rates of dry weight gain and activity of sucrolytic enzymes. Once anther growth was completed, the dry weight gain of tepal, filament, stigma and style increased with a concomitant increase in hexose concentrations and invertase activity. Although all three enzymes capable of catalyzing sucrose cleavage were present in every flower organ of L. longiflorum , soluble invertase was the predominant enzyme in all flower organs except stigma where cell wall invertase dominated. Soluble invertase activity was highly correlated with dry weight gain in most of the flower organs.     

Early effects of excess cadmium uptake in Phaseolus vulgaris

Abstract. Seedlings of Phaseolus vulgaris were exposed to solutions containing Cd²⁺ in the range 0 to 1 molm⁻³. Ethylene formation started following 3 h of exposure to 10⁻², 10⁻¹ and 1 mol m⁻³ Cd²⁺, peaked at 18 h and returned to a relatively low rate after 24 h. Cadmium-induced ethylene formation depended on the formation of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminoethoxyvinylglycine (AVG, 0.1 mol m⁻³) inhibited ACC accumulation and ethylene production during exposure to 0.2 mol m⁻³ Cd²⁺.
Activity of soluble and ionically-bound peroxidase increased after 18 h of exposure to Cd²⁺ concentrations above 10⁻³ mol m⁻³ due to an increase in activity of cathodic isoperoxidases. Stimulation of soluble and ionically-bound peroxidase by 0.2 mol m⁻³ Cd²⁺ was reduced in the presence of 0.1 mol m⁻³ AVG.
Accumulation of soluble and insoluble ('ligninlike') phenolics was found in plants exposed to Cd²⁺ (10⁻² mol m⁻³ or above) in the presence or absence of AVG. Deposition of insoluble (autofluorescing) material occurred in cell walls around vessels and was associated with reduced expansion and water content of leaves.     

Photosynthetic characteristics of the submerged macrophyte Ceratophyllum demersum

The photosynthetic and growth characteristics of Ceratophyllum demersum L. were investigated under laboratory conditions which simulated those encountered in the plants' normal environment. The carbon fixation rate of C. demersum measured with ¹⁴C at light and carbon saturation at pH 8.0 was 4.48 mg C (g ash-free dry weight)⁻¹ h⁻¹. It was lower at pH 6.5 than at pH 8.0. The light use efficiencies in quiescent plants and actively growing plants were 6.3 and 8.7 × 10⁻⁹ kg CO₂ J⁻¹, respectively, with corresponding maximum photosynthetic rates of 2.67 and 4.36 mg C (g ash-free dry weight)⁻¹ h⁻¹. Photorespiration in actively growing plants consumed 24% of the carbon fixed. Incubation with DCMU demonstrated that about one-third was refixed. The optimum temperature for carbon fixation was 25°C. The C₃-photosynthetic pathway was the main operational route as indicated by the early photosynthetic products (largely C₃-acids) and the absence of Krantz anatomy and the chlorophyll a:b ratio (2.7). The maximum relative growth rates ranged from 0.025 to 0.041 g ash-free dry weight (g ash-free dry weight)⁻¹ day⁻¹ in the field (Lake Vechten, 1 to 3 m depth classes).     

Seed dimorphism in Salicornia europaea: Nutrient reserves

Median and lateral seeds of Salicornia europaea L. were separately analysed for their sizes and nutrient reserves. The mean air-dry weight of a single median and lateral seed was 0.31 and 0.25 mg, respectively. The composition as well as the concentration of the nutrient reserves were similar in both seed types. The bulk of the cations was derived from K⁺, followed by Mg²⁺, Na⁺ and Ca²⁺. The chloride content was somewhat higher than the sodium content, and phosphate was equalled by acid soluble Ca²⁺ and Mg²⁺. Starchy compounds and sucrose were present in equal amounts, each of them accounted for about 50% of the carbohydrates. Glucose and fructose were less than 1%. Protein-nitrogen (ethanol-insoluble N) was about 34 g (kg dry seeds)⁻¹. About 7 g (kg dry seeds)⁻¹ was ethanol-soluble nitrogen, of which 10% was derived from amino acids. The total lipid content was more than 290 g (kg dry seeds)⁻¹, 65% were calculated to be glycerides. More than 90% of the fatty acids consisted of linoleic and oleic acids, the majority (72%) of which was linoleic acid.     

Changes in diffusible indole-3-acetic acid from various parts of tulip plant during rapid elongation of the flower stalk

In order to chemically identify the putative indole-3-acetic acid (IAA) and to confirm the native source of auxins account for rapid elongation of the floral stalk of tulip, we examined diffusible IAA from various parts of tulip plant during rapid elongation of the flower stalk. IAA was identified in the diffusates collected from the leaves, internodes, and floral organs with gas chromatography (GC)–mass spectrometry. The amount of diffusible IAA from different plant organs followed the order of that the internodes > flower organs > leaves during the period of rapid elongation of the floral stalk. The diffusible IAA from internodes reached its peak amount at different time than did diffusible IAA from the flower. The results obtained indicated that the top internode is probably the major source of auxins account for rapid elongation of the flower stalk.     

Respiratory metabolism during cold storage of apple fruit. II. Alternative oxidase is induced at the climacteric

Rubbing applied to a young tomato internode inhibited the elongation of this internode and increased soluble peroxidase activity. These morphological and biochemical changes were observed both at the site of rubbing (local response) and in the neighbouring internode (systemic response). The cellular, biochemical, and molecular mechanisms leading to inhibition of internode elongation are not fully understood. It was proposed that mechanical stimuli increased the oxidation of IAA, via the induction of specific peroxidases and stimulated the lignification processes. In order to gain more information about the role of these enzymes, analysis of changes in peroxidase activities were performed. Qualitative analysis of isoperoxidases, by means of native cathodic PAGE, showed four induced isoforms termed C₁, C₂, C₃, and C₄. The major isoform (C₂) was purified to homogeneity and partially characterized. This isoform is probably unglycosylated, with a molecular mass of 36 kDa and a neutral pI of 7.1. The effects of pH and temperature on the activity were determined with guaiacol as electron donor. Optima were obtained at pH 5 and at a temperature of 55°C. The activity of the purified enzyme was not affected by Ca²⁺, Mg²⁺ and Mn²⁺ as was reported for some basic peroxidases. Analysis of substrate specificity revealed that this isoperoxidase acted on ABTS, o -dianisidine, pyrogallol, guaiacol, coniferyl alcohol (monolignol) and IAA but not on syringaldazine. Activitiy of C₂ isoperoxidase on coniferyl alcohol and IAA suggests a possible role of peroxidase C₂ in inhibition of internode elongation, observed in rubbed plants, probably via an increase in lignification processes and regulation of IAA levels in internode tissues.