Resumption of cellular activity induced by cytokinin and gibberellin treatments in tomato flowers targeted for abortion in unfavorable light conditions

Mitotic activity and nuclear DNA synthesis in tomato ( Lycopersicon esculentum Mill., cv. King plus) flowers targeted for abortion under unfavorable light conditions are completely stopped 6 days after macroscopic appearance of the inflorescence. Ovular cells are arrested at the G₁ (80%) and G₂ (20%) stages of the cell cycle. Exogenous applications of a mixture of N⁶-benzyladenine (BA) and gibberellins A₄₊₇ (GA) directly on the inflorescence may prevent its failure. Nuclear DNA synthesis and mitoses resume in ovules of the flower 16 to 20 h after the BA+GA treatment. When applied alone, BA and GA are able to mimic the effect of the mixture upon the progression of ovular cells through their cycle. Sporogenesis processes are also set in motion by the exogenous plant growth regulators. The mechanism of action of cytokinins and gibberellins in the control of floral development is discussed.     

[14C]-Assimilate partitioning in photoperiodically induced seedlings of Pharbitis nil. The effect of benzyladenine

Partitioning of [¹⁴C]-labeled assimilates was studied in relation to photoperiodic floral induction and evocation in one-week-old Pharbitis nil Choisy cv. 'Violet' seedlings. In plants kept under 16 h photoperiods, one 15 h night induced 100% axillary flowering whereas a 24 h night induced both terminal and axillary flowering. A 15 min night break of red light given 8 h after the beginning of the dark period inhibited flowering. Total [¹⁴C]-assimilate distribution among major sinks (plumules + epicotyl and roots + hypocotyl) from a single source cotyledon was unchanged by one inductive night; however, import of [¹⁴C]-assimilates into shoot apices was increased in induced plants compared to vegegative controls. This increase was several-fold in plants subjected to a 24 h night. N⁶-Benzyladenine (BA) application to cotyledons or plumules under non-saturating night lengths increased the number of floral buds per plant without affecting the position of the first floral bud (i.e. the speed of induction). The same treatment caused increased label accumulation in induced apices, while it only slightly affected non-induced ones. The mode of action of BA on flowering through growth stimulation and resulting assimilate mobilization is discussed.     

Retranslocation of boron in broccoli and lupin during early reproductive growth

The objective of the present study was to determine if boron (B) retranslocation depends on plant-B status and external-B supply. The stable ¹⁰B isotope was supplied to the root system of broccoli ( Brassica oleracea var. italica Plenck cv. Commander) and lupin ( Lupinus albus L. cv. Ultra) plants to provide a quantitative picture of B distribution during early reproductive development. Regardless of the B regime (i.e. continuous supply with luxury, sufficient or deficient B; transfer at inflorescence emergence from either a luxury- or sufficient-B supply to a deficient one) and whether ¹⁰B was acquired before or during inflorescence development, a significant proportion of the B recovered in broccoli florets and lupin fruit was ¹⁰B enriched. B acquired during inflorescence development was an important source of B for reproductive structures, but the relative importance of B acquired before and after inflorescence emergence appeared to be species dependent. The occurrence of B retranslocation was not dependent upon the induction of B deficiency. The concentrations of B in phloem exudates (0.38 to 0.03 mM) were 4- to 23-fold those in xylem sap, and more similar to the concentrations in the reproductive structures (0.86 to 0.07 mM) than those in source leaves (2.4 to 0.19 mM). The decreasing acropetal gradient of tissue-B concentrations with luxury-B supply declined dramatically or was reversed in plants grown with sufficient or deficient B. The data are consistent with B being a phloem-mobile element, and suggest that newly acquired B is particularly important during the early reproductive growth of plants.     

Stimulation of shoot elongation in Salix pentandra by gibberellin A9; activity appears to be dependent upon hydroxylation to GAl via GA20

Cessation of shoot elongation in seedlings of Salix pentandra L. is induced by short photoperiod. Gibbereliin A₉ (GA₉) applied either to the apical bud or injected into a mature leaf, induced shoot elongation under a short photoperiod of 12 h, and GA₉ could completely substitute for a transfer to a long photoperiod. When [³H]GA₉ or [²H₂]GA₉ was injected into a leaf, no [³H]GA₉ was detected in the elongating apex and only traces of [³H]GA₉ were found in the shoot above the treated leaf. By the use of gas chromatography-mass spectrometry (GC-MS), [²H₂]GA₂₀ was identified as the main metabolite of [²H₂]GA₉ in both the shoot and the treated leaf. In addition, [²H₂]GA₁ and [²H₂]GA₂₉ were also identified as metabolites of [²H₂]GA₉. These results are consistent with the hypothesis that exogenous GA, promotes shoot elongation in Salix through its metabolism to GA₂₀ and GA,.     

Translocation of labelled assimilates following photosynthesis of 14CO2 by the field bean, Vicia faba

When whole plants were exposed to ¹⁴CO₂, almost the same amount of radioactivity was taken up initially by each leaf regardless of its position on the stem and of the presence of beans at that node. Thus, although developing beans are a powerful sink for assimilated carbon, they do not increase the CO₂ uptake by adjoining leaves.
The distribution of labelled assimilates 6 hours after feeding ¹⁴CO₂ to a single leaf for 1 hour varied with both the position of the treated leaf and the stage of development of the plant. Before any flowers were set most of the radioactivity from all expanded leaves moved downwards to the roots and the stem below the treated leaf (lower stem). Later, during pod-fill, the upper leaves maintained this supply to the roots and lower stem, whilst most of the carbon translocated from the lower and mid-stem leaves went to the beans. However, we found no exclusive relationship between a leaf and the supply to beans developing on the same node.
The amount of radioactivity moving out of a source leaf at a fruiting node increased over successive samplings up to 48 h; the pattern of distribution of the ¹⁴CO₂ however remained virtually unchanged.     

Effect of cytokinin on the distribution of 14C-glyphosate in Xanthium pennsylvanicum

Cytokinin N⁶-(δ²-isopentenyl) adenosine (i⁶A) was applied five times to runoff on the first leaf of Xanthium pennsylvanicum Wallr. After a 24 h application period the second leaf was treated with the herbicide methyl-¹⁴C-glyphosate, [N-(phosphonomethyl)glycine]. The distribution of radioactivity was determined after 12 and 24 h. At no time after treatment did ¹⁴C-glyphosate move preferentially into the i⁶A treated leaf. Radioactivity accumulated in all plant parts, but in i⁶A-treated plants, there were more counts in the actively growing areas after 12 h. The primary effect of i⁶A after 24 h was enhanced uptake of ¹⁴C-glyphosate into the plant. To determine if the increased uptake and altered movement of ¹⁴C-glyphosate was due to a cytokinin-induced transpiration increase, leaf diffusive resistance was measured. Leaf diffusive resistances were unaffected by i⁶A during the 48 h after application. By 12 h after the application of glyphosate, the herbicide was found not to affect the cytokinin content in the Xanthium root tips.     

Effect of plant density on the uptake and distribution of 14C in the field bean, Vicia faba

Whole bean plants, ev. Cockfield, grown in pots crowded or well-spaced (50 or 10 plants m², respectively) were treated with ¹⁴CO₂ at the pod-fill stage (25 modes) and the radioactivity in each leaf was determined after 30 min. With spaced plants the uptake was greatest in the mid-stem leaves and was proportional to leaf area. In contrast, 70% of the total assimilation took place in the upper six leaves of crowded plants and there was a steady decrease down the stem.
When ¹⁴CO₂ was fed to single leaves of similar crowded plants the resultant distribution of labelled assimilates varied with the position of the treated leaf. After 6 h, 67% of the ¹⁴C fixed by a mid-stem leaf (node 13) was recovered from the beans, whereas 76% of that from an upper leaf (node 23) had accumulated along the stem. Due to the shading of mid-stem leaves at the higher planting densities, seed yield becomes increasingly dependent upon re-distribution of assimilates from stem to beans.     

Distribution of 14C-sucrose applied to leaves at different nodes of okra (Abelmoschus esculentum) during flowering and early pod growth

The distribution pattern of ¹⁴C-sucrose from ¹⁴C-sucrose applied to vegetative okra plants and leaves 1–9 on separate plants during the green pod development stage were investigated in relation to duration and leaf position. Results indicated bi-directional transport of assimilates to both apical and basal portions of the stem. Within 48 h ¹⁴C moved to all plant parts; stem and leaves appeared to be strong sinks. In plants fed at the vegetative stage, 48 h after feeding, 66% of the fed activity was exported from the fed leaf. At the pod development stage, about 35% of the activity exported from the fed leaf was present in green pods and 65% in vegetative parts. In plants where leaf 1–9 was fed, irrespective of the position of the fed leaf, the subtending fruit was the strongest sink among the reproductive parts. Leaves and stems were the principal sinks.     

Translocation of photoassimilate from leaves of two popyploid genotypes of tall fescue differing in photosynthetic rates

The photosynthetic rate of a decaploid genotype (1-16-2) of tall fescue ( Festuca arundinacea Schreb.) is about twice that of a common hexaploid genotype (V6-802) (Plant Physiol. 72: 16–21, 1983). Translocation of photosynthate out of the leaves is a possible means of regulating carbon assimilation. To evaluate this possibility, we have examined a) translocation velocity, b) time course of translocation from leaves, c) photoassimilate partitioning pattern into whole plants in pulse and chase experiments, and d) interveinal distances between two ploidy genotypes. Most of the ¹⁴C accumulated in sucrose, and the labelled carbon moved down the leaf blades at similar velocities (6 to 10 cm h⁻¹) in both genotypes. Recent ¹⁴C assimilate was rapidly translocated from the fed area of the leaf blade. For example, the decaploid and the common hexaploid had translocated 40 and 26% of the ¹⁴C, respectively, at 6 h, and 79 and 49% of the ¹⁴C, respectively, at 24 h. Partitioning of ¹⁴C among plant organs was considerably different between the genotypes after a 24 h chase. For example, out of the total ¹⁴C recovered from the whole plant, the decaploid had retained 40% in the labelled leaf with 10, 33 and 29% in other leaves, stem bases and roots, respectively; whereas the hexaploid had retained 91% in the labelled leaf with 4, 3 and 2% in other leaves, stem bases and roots, respectively. However, the higher rate of translocation was correlated with greater interveinal distances in the decaploid genotype. These results suggested that the higher translocation percentage in the decaploid than the hexaploid genotype was due to greater sink activity.     

The relationship between photosynthetic electron transport and photorespiratory 14CO2 release after DCMU treatment in the duckweed, Lemna gibba

The photosynthetic rate of Lemna gibba was measured as ¹⁴CO₂ uptake at the beginning of and after 1 h DCMU treatment during the separate excitation of PS I (703 nm), mainly PS II (662 nm) and the combined excitation of both photosystems (662 + 703 nm) in 2 and 21% oxygen. The results show the Warburg effect. Photosynthesis was significantly reduced by DCMU whenever PS II was excited, at 662 nm and 662 + 703 nm. Photosynthetic enhancement was greater in 21 than in 2% oxygen in both the treated and untreated plants.
Photorespiratory ¹⁴CO₂ release was only affected by DCMU treatment at 662 + 703 nm. It was significantly decreased in 21% O₂ and significantly increased in 2% O₂ as compared to the controls without DCMU. The ¹⁴C-glycolate remaining in the plant after photosynthesis/photorespiration measurements was reduced whenever the electron supply to PS I was low.
These data support the hypothesis that a relationship exists between glycolate metabolism and photosynthesis via the electron transport chain where electrons from the oxidation of glycolate are donated to PS I when the electron supply from water is low.     

Altered resource allocation during seed development in Arabidopsis caused by the abi3 mutation

The regulation of whole-plant resource allocation during seed development in Arabidopsis thaliana was investigated by examining growth rate and partitioning of ¹⁴CO₂ in wild-type plants and those carrying the abi3 mutation. Plants carrying the abi3 mutation partitioned more resources into seed development than the wild type. The extra resources were available as a result of delayed senescence of the cauline leaves in the mutant. After supply of ¹⁴CO₂ at later stages of reproductive development differences in patterns of ¹⁴C distribution between mutant and wild type were consistent with long-term changes in growth and allocation. The role of long-distance signals in the regulation of seed yield in Arabidopsis is discussed.     

The trans-tissue pathway and chemical fate of 14C photoassimilate in carrot taproot

Axial and radial transport and the accumulation of photoassimilates in carrot taproot were studied using ¹⁴C labelling and autoradiography. Axial transport of the ¹⁴C labelled assimilates inside the taproot was rapid and occurred mainly in the young phloem found in rows radiating from the cambium. The radial transport of the assimilate inward (to cambium, xylem zone and pith) and outward (to phloem zone and periderm) from the conducting phloem was an order of magnitude slower than the longitudinal transport and was probably mainly diffusive. The cambial zone of the taproot presented a partial barrier in the inward path of the assimilate to the xylem zone. We suggest that this is due to the cambium comprising a strong sink for the assimilate on the basis that our previous work has shown that it contains very low concentrations of free sucrose. By contrast, a high accumulation of nonsoluble ¹⁴C was found in the cambium region in good agreement with the active growth of this zone. Autoradiography following the feeding of ¹⁴C labelled sugars to excised sections of taproot indicated that only a ring of cells at and/or just within the cambium take up sugars from the apoplast. This indicates that radial movement in the phloem and pith must be symplastic. An apoplastic step between phloem and xylem is possible. The rapid uptake of sugars from the apoplast at this point might represent a mechanism for keeping photoassimilates away from the transpiration stream and re-location back to the leaves.     

Does light-promoted export from Commelina benghalensis leaves result from differential light-sensitivity of the cells in the mesophyll-to-sieve tube path?

In contrast to the light-promoted uptake by mesophyll cells, light slightly inhibited sucrose uptake by stripped leaf disks of Commelina benghalensis L. This phenomenon appeared to result from a light-promoted vein-associated release, as light stimulated photosynthate release from stripped disks and inhibited that from mesophyll cells. In the -presence of the resorption-blocker p -chloromercuriphenylsulfonic acid, (PCMBS) the release of preloaded [¹⁴C]-sugars (sucrose, glucose) and [¹⁴C]-amino acids (alanine, asparagine, proline, valine, α-aminoisobutyric acid) from stripped disks was doubled in the light. Illumination enhanced by 20 to 60% the release of endogenous leaf cell compounds (sucrose, H₂PO-₄, K⁺, Mg²⁺, Ca²⁺) from stripped disks in the presence of PCMBS. Light also increased the export of [¹⁴C]-assimilates from intact leaves by 20% after pulse-labelling with ¹⁴CO₂. A model for loading is proposed, based on the differential light sensitivities of the plasma membranes in the mesophyll-to-sieve tube path.     

Effects of gibberellic acid on patterns of carbohydrate distribution and acid invertase activity in Phaseolus vulgaris

The application of gibberellic acid (GA₃,10 μ M ) as a root drench to 16-day-old plants of Phaseolus vulgaris L. cv. Masterpiece stimulated growth of the stem internodes and reduced root growth. GA₃ treatment did not affect the export of ¹⁴C from a primary leaf to which [¹⁴C]-sucrose was applied, but greatly increased upward translocation to the elongation region of the stem at the expense of transport to the hypocotyl and root system. The observed changes in the patterns of growth and [¹⁴C]-labelled assimilate distribution were correlated with an increase in the specific activity of acid invertase in the elongating stem internodes and a decrease in invertase activity in the hypocotyl and root. Sucrose concentration in the elongating internodes fell substantially after treatment with GA₃ while the concentration of hexose sugars increased. We suggest that by stimulating acid invertase synthesis in the elongating internodes, GA₃ acts to establish a more favourable sucrose gradient between these sinks and source leaves. Under source-limiting conditions this, in turn, will lead to a reduced rate of assimilate translocation to competing sinks in the root system.     

Growth and accumulation of N, K+, Ca2+ and Mg2+ in barley exposed to various nutrient regimes and root/shoot temperatures

Six cultivars of spring barley ( Hordeum vulgare L. cvs Salve, Nümberg II, Bomi, Risø 1508, Mona and Sv 73 608) were grown in water culture for three weeks with various combinations of mineral supply and differential roots/shoot temperatures during the growth period. Most important for growth and accumulation of N, K⁺, Ca²⁺ and Mg²⁺ was the mineral supply, followed by the root temperature and the choice of cultivar. Treatments with low mineral supply or low root temperature induced a uniform reduction in growth and accumulation of the ions studied. The effects of low mineral supply and low root temperature on growth and N accumulation was additive, which indicates that these factors exert their influence independently of each other.
Roots grown at 10°C were smaller and Rb⁺(⁸⁶Rb) influx was higher than in roots grown at 20°C. It is suggested that the control of Rb⁺(⁸⁶Rb) influx is affected by the root temperature and the age of the plants. The higher ⁸⁶Rb⁺ (⁸⁶Rb) influx into the low temperature roots could not compensate for the smaller root size. However, the lower total mineral accumulation made up for the needs of the smaller plants and cannot explain the reduction in growth.     

Comparative studies of sucrose and mannitol utilization in celery (Apium graveolens)

Utilization of sucrose and mannitol, the major forms of translocatable assimilate in celery ( Apium graveolens L. cv. Giant Pascal), was investigated in intact plants, excised leaves and leaf discs by estimating the soluble carbohydrate pools, starch levels and oxidation of [¹⁴C]-sucrose or mannitol in the light and after extended dark treatments. In detached mature fully-expanded leaves, mannitol pools remained constant, while sucrose decreased during a 48 h dark treatment. In attached leaves on plants trimmed to a single compound leaf, however, mannitol levels decreased after a dark treatment. In leaf discs floated on bathing solutions containing [¹⁴C]-sucrose or [¹⁴C]-mannitol, oxidation of mannitol was restricted to young leaf tissues, whereas sucrose was metabolized to CO₂ regardless of leaf age. Uptake of labelled mannitol, however, was greater than that of sucrose in the light in leaves of every age. Although both mannitol and sucrose are translocated out of leaf tissues, leaf age differences indicate that, unlike sucrose, mannitol utilization is restricted to active sink tissues. The results suggest different roles for mannitol and sucrose with mannitol representing a more rigorously sequestered transport carbohydrate.     

Accumulation and Spatial Distribution of Poly(A)+RNA in Oocytes and Early Embryos of Drosophila melanogaster

We describe the accumulation and distribution of poly (A)⁺RNA during oogenesis and early embryogenesis as revealed by in situ hybridization with a radio-labeled poly (U) probe. The amount of poly (A)⁺RNA in nurse cell cytoplasm continuously increased untill mid-vitellogenic stage (st. 10), then decreased with the rapid increase of poly (A)⁺RNA in the oocyte (st. 11). The localization of poly (A)⁺RNA at stage 10 was in the anterior region of the oocyte, where it is connected by cytoplasmic bridge to the nurse cells. These observations indicate that most of the poly (A)⁺RNA synthesized in the nurse cells is transferred to the oocyte through the cytoplasmic bridges at stage 10–11. During the remainder of oogenesis (st. 11–14) and during preblastodermal embryogenesis, poly (A)⁺RNA was evenly distributed over the cytoplasm of oocytes and embryos. At blastoderm stage, poly(A)⁺RNA became concentrated in the peripheral region of embryos. Though the somatic nuclei of the blastoderm contained a detectable amount of poly (A)⁺ RNA, the pole cell nuclei did not. The cytoplasmic RNA visualised by acridine orange staining and the poly (A)⁺RNA detected by hybridization with [³H]poly (U) exhibited identical distributions during oogenesis and early embryogenesis. These observations provide a basis to assess the unique distributions of specific RNA sequences involved in early development.     

Distribution of 14C assimilates in the storage root of sugar beet plants after import from leaves of different age

In the sugar beet plant ( Beta vulgaris L. ssp. altissima ) the vascular bundles of old leaves lead to the center and those of young leaves to the periphery of the storage root. Whether the flux of assimilates follows these anatomical routes was tested by applying ¹⁴CO₂ for 4 h to either an old (10th) or a young (20th) leaf in intact sugar beet plants. Four-month-old plants, which had about 30 leaves, were used in the experiment. The ¹⁴C distribution in the storage root was measured by autoradiography and counting in about 20 cross and longitudinal sections per root.
About 37% of assimilated ¹⁴C from an old leaf and 23% from a young leaf were exported within 24 h. Although some ¹⁴C moved into younger leaves, most was exported into the storage root. During its rapid movement towards the root tip, which took place perferentially in the orthostichon belonging to the [¹⁴C]-treated leaf, the label spread laterally.
The autoradiograms indicate that the distribution of assimilates within the storage root is roughly determined by the course of the vascular bundles extending from the source leaf. The fine distribution, however, seems to be controlled by sucrose gradients between storage cells.     

Ca2+ UPTAKE BY SYNAPTOSOMES AND ITS EFFECT ON THE INHIBITION OF ACETYLCHOLINE RELEASE BY BOTULINUM TOXIN

Abstract— ⁴⁵Ca²⁺ uptake by cerebral cortex synaptosomes was determined by gel filtration, glass fibre disc filtration under suction and by centrifugation with EGTA present. The filtration methods gave comparable results which were higher than values obtained by the centrifugation method. Uptake was increased by 25mM-K⁺ at all times investigated. The accumulated ⁴⁵Ca²⁺ was bound within the synaptosome. ⁴⁵Ca²⁺-ionophore A23187 stimulated uptake only during the first min; levels of intra-synaptosomal ⁴⁵Ca²⁺ then returned to control values. A23187 also increased intra-synaptosomal Na⁺ and Cl⁻ contents. Botulinum toxin inhibits the K.⁺-stimulated release of [¹⁴C]ACh from synaptosomes but the ionophore released [¹⁴C]ACh from both normal and botulinum-treated preparations in a Ca²⁺-dependent manner. However, it also elicited Ca²⁺-dependent release of [choline. Increased extracellular Ca²⁺ (10 mM and 20 mM) released [¹⁴C]ACh (but not [¹⁴C]choline) from both normal and botulinum-treated synaptosomes. It is concluded that botulinum toxin interferes with the provision of Ca²⁺ essential for the mechanism of ACh release.     

Vitamin D3 affects growth and Ca2+ uptake by Phaseolus vulgaris roots cultured in vitro

Vitamin D₃ at low concentration (10⁻⁹ M) inhibited the growth of Phaseolus vulgaris L. (cv. Contrancha) roots in vitro as measured by elongation (14 h) and [³H]-leucine incorporation into protein (2 h), and increased their labelling with ⁴⁵Ca²⁺ (2 h). Cycloheximide and puromycin (50 u.M) blocked vitamin D₃ stimulation of root ⁴⁵Ca²⁺ labelling, indicating that it is mediated by de novo protein synthesis. The calcium ionophore X-537A (10⁻⁵JW) induced similar changes both in root elongation and ⁴⁵Ca²⁺ uptake (14 h). This may indicate that the inhibitory effects of the sterol on root growth are mediated by changes in Ca²⁺ fluxes. However, this interpretation should be further strengthened by additional studies as the ionophore may have acted on root growth, affecting physiological processes other than Ca²⁺ transport.