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The role of strigolactones as plant growth regulators has been demonstrated through research on biosynthesis and signaling mutant plants and through the use of GR24, a synthetic analog of this class of molecules. Strigolactone mutants show a bushy phenotype and GR24 application inhibits the growth of axillary buds in these mutants, thus restoring the phenotype of a wild plant, which is characterized by a stronger apical dominance. In this work, we tested the effectiveness of this chemical on pea (Pisum sativum) plants following apex removal, which disrupts apical dominance and leads to axillary bud outgrowth. Moreover, we searched for relationships between the response to the strigolactone and gibberellin metabolism by applying GR24 to both climbing and dwarf peas, the latters being mutants for gibberellin biosynthesis. The results suggest that the endogenous level of the bioactive gibberellin GA1 might modulate the response of decapitated pea plants to GR24, by changing bud sensitivity to the applied strigolactone.

The vegetation profile and the photosynthetic efficiency, oxidative damage, and stomatal conductance in the evergreen dwarf shrub Calluna vulgaris (L.). Hull was analyzed in a Mediterranean ecosystem characterized by intense geothermal activity. Among the higher plants present in the area, this species appears to be the sole to possess the ability to grow near the geothermal sources. The hot fluid springs strongly alter the environment in their proximity: the emitted water vapor, CO2, H2S, CH4, H3BO3, SO4 2−, and NH4 + partly condensate and precipitate to the soil, thus leading to its extreme acidification and nutrient depletion. Furthermore, the temperature starts to rise sharply just a few centimeters under the soil surface. Under this multiple stress, the individuals of C. vulgaris growing within a few meters from the springs showed lower photosystem II efficiency, higher oxidative damage to the biomembranes, and lower stomatal conductance than the individuals growing farther away. Drought and high air temperatures occurring in summer exacerbate these harsh conditions, but only the plants closer to the springs did undergo an acute, yet transient crisis, as shown by the analyzed parameters. These results suggest that the main factors of stress are related to the physical and chemical features of the soil, while the adverse climate conditions apparently are of secondary importance. The possible role of reduced stomatal conductivity in enhancing the resistance of C. vulgaris to this hostile environment is discussed.

Summary In this paper we analysed the endogenous IAA levels in carrot cell cultures and in cell culture medium. Free IAA and its conjugates (esters and amides) were measured in cell cultures exposed to different concentrations of 2,4-D (nil, 2,3 and- 92 μmol/L). The three culture conditions determine different physiological conditions of the cell cultures: somatic embryogenesis, unorganised growth and growth under stress, respectively. The results indicate that these two types of IAA conjugates account for 60–70 % of total IAA and buffer the variations of free IAA in a sequential manner. Free IAA is in a steady state relationship with the ester conjugates and the increased synthesis of IAA is initially buffered by IAA-amide. When a certain threshold (750 ng/g) is reached inside the cell by the amide, further increments are buffered by ester IAA. Another result coming from this analysis is that the presence of tryptophan alone in the culture medium is sufficient to sustain, metabolically as well as physiologically, the conditions of proliferation, shifting the state of the cell culture from somatic embryogenesis to unorganised growth.

Endosperm of Sechium edule seed was incubated with [U-14C]adenine and [13C]mevalonic acid for 17 h. Following incubation, cytokinins were extracted and purified by HPLC. Using GC-MS, incorporation of [13C]MVA into isopentenyladenine and zeatin riboside was demonstrated. This is the first unequivocal demonstration of cytokinin biosynthesis in a normal plant tissue because precursors incorporation into cytokinins was demonstrated by MS. The significance of these results is also discussed in relation to a possible mononucleotide-independent pathway of cytokinin biosynthesis.

Senescence is a tightly regulated process and both compartmentalisation and regulated activation of degradative enzymes is critical to avoid premature cellular destruction. Proteolysis is a key process in senescent tissues, linked to disassembly of cellular contents and nutrient remobilisation. Cysteine proteases are responsible for most proteolytic activity in senescent petals, encoded by a gene family comprising both senescence-specific and senescence up-regulated genes. KDEL cysteine proteases are present in senescent petals of several species. Isoforms from endosperm tissue localise to ricinosomes: cytosol acidification following vacuole rupture results in ricinosome rupture and activation of the KDEL proteases from an inactive proform. Here data show that a Lilium longiflorum KDEL protease gene (LlCYP) is transcriptionally up-regulated, and a KDEL cysteine protease antibody reveals post-translational processing in senescent petals. Plants over-expressing LlCYP lacking the KDEL sequence show reduced growth and early senescence. Immunogold staining and confocal analyses indicate that in young tissues the protein is retained in the ER, while during floral senescence it is localised to the vacuole. Our data therefore suggest that the vacuole may be the site of action for at least this KDEL cysteine protease during tepal senescence.

Summary Tubers cv. Monalisa were analysed to determine the content of abscisic acid (c-t-(+)ABA) and related metabolites in three tuber parts (eyes, sub-eye tissues and pith). Analyses were performed from the last period of tuber growth until sprouting under two different storage temperatures (3 °C and 23 °C). ABA content rises in eyes and sub-eye tissues as sprouting approaches, regardless of storage temperature. Dihydrophaseic acid is the main ABA metabolite while phaseic acid was not detected, probably because of its further metabolization to DPA. ABA-glucose ester concentrations are generally low in all tissues except in eyes before harvest. The results are discussed in relation to the current hypotheses on the role of ABA in the control of potato tuber dormancy.

The nucellus is a maternal tissue that feeds the developing embryo and the secondary endosperm. During seed development the cells of the nucellus suffer a degenerative process early after fertilization as the cellular endosperm expands and accumulates reserves. Nucellar cell degeneration has been characterized as a form of developmentally programmed cell death (PCD). In this work we analysed the role of the endosperm as main regulator of nucellus PCD. We demonstrated that endosperm produces high amount of ethylene, nitric oxide and indoleacetic acid. We examined the role of these small and diffusible signalling molecules in the regulation of nucellus PCD and we tried to elucidate how they can cooperate and regulate each other into the endosperm. We showed that ethylene acts a positive regulator of nucellus PCD and its synthesis can be in part induced by nitric oxide. High levels of IAA were detected both in the endosperm and in dying nucellus but this hormone is not directly involved in the execution of PCD.

Grains of wheat ( Triticum durum Desf. cv. Cappelli) were collected at different stages of maturation. To test whether the response to desiccation injury is correlated with physiological stage and/or water content, one lot of grains at each stage was artificially dried until its water content was comparable to that of the subsequent stage. Several stress parameters were analysed in embryos isolated from naturally-and artificially-dried grains. In particular, the content of dehydrins, hydrogen peroxide and the activity of ribonucleases and of antioxidant enzyme were studied. Neutral ribonucleases, unlike acidic ones, seemed to be correlated with grain water content. In parallel with these analyses the presence of dehydrins was also examined. Dehydrin accumulation was found to vary with drying rate, and more isoforms of dehydrin were present in artificially dried than in naturally-dried grains. Artificial drying resulted in accumulation of hydrogen peroxide, while the content of this molecule was much lower in embryos from naturally fully-ripened grains. While the activities of ascorbate peroxidase (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1), glutathione reductase (GR, EC 1.6.4.2), and glutathione peroxidase (GPX, EC 1.11.1.9), were generally higher early in maturation, the activity of catalase (CAT, EC 1.11.1.6) increased in mid-maturation and decreased in fully-ripe embryos. Different protective mechanisms seem to act in combination but their relative importance changes as maturation progresses and in response to the drying method.

The last phase of flower development is senescence during which nutrients are recycled to developing tissues. The ultimate fate of petal cells is cell death. In this study we used the ethylene-insensitive Lilium longiflorum as a model system to characterize Lily flower senescence from the physiological, biochemical and ultrastructural point of view. Lily flower senescence is highly predictable: it starts three days after flower opening, before visible signs of wilting, and ends with the complete wilting of the corolla within 10 days. The earliest events in L. longiflorum senescence include a fall in fresh and dry weight, fragmentation of nuclear DNA and cellular disruption. Mesophyll cell degradation is associated with vacuole permeabilization and rupture. Protein degradation starts later, coincident with the first visible signs of tepal senescence. A fall in total protein is accompanied by a rise in total proteases, and also by a rise of three classes of caspase-like activity with activities against YVAD, DEVD and VEID. The timing of the appearance of these caspase-like activities argues against their involvement in the regulation of the early stages of senescence, but their possible role in the regulation of the final stages of senescence and cell death is discussed.

The nucellus is a maternal tissue that feeds the developing embryo and the secondary endosperm. During seed development the cells of the nucellus suffer a degenerative process early after fertilization as the cellular endosperm expands and accumulates reserves. Nucellar cell degeneration has been characterized as a form of developmentally programmed cell death (PCD). In this work we show that nucellus PCD is accompanied by a considerable production of both nitric oxide and hydrogen peroxide (NO and H(2)O(2)). Interestingly, each of the two molecules is able to induce the production of the other and to cause cell death when applied to a living nucellus. We show that the induced cell death has features of a PCD, accompanied by profound changes in the morphology of the nuclei and by a massive degradation of nuclear DNA. Moreover, we report that NO and H(2)O(2) cause an induction of caspase-like proteases previously characterized in physiological nucellar PCD.

Summary The relationship between fruit and seed tissues and plantlet growth was investigated in Sechium edule Sw. Removal of fruit and seed teguments showed no effect on plantlet growth while the removal of cotyledons arrested growth under short days. On the contrary, the plantlets continued to grow actively after excision of cotyledons under long day conditions. Plantlets sprayed daily with a 5 x 10 −5 M solution of GA 4/7 , after cotyledon removal did not experience a growth arrest under short day conditions but GA 4/7 supplied through the petiole in more physiological amounts failed to compensate for the excised cotyledons. The gibberellin glucoside (GAG) that accumulates in the seed during development was extracted, purified and applied to cotyledon-deprived plantlets. While a resumption in growth was not induced in the apical shoot the treatment did stimulate the active growth of secondary shoots. It is suggested that the control of early plantlet growth by cotyledons under short day conditions may be mediated by GAs.

Plant growth regulators are involved in the control of potato (Solanum tuberosum) tuber dormancy. Evidence concerning the role of IAA is controversial; we therefore investigated its role by analyzing two cultivars with varying lengths of dormancy. We examined the time course of free and conjugated IAA in tuber tissue isolates from the final stages of tuber growth to the end of dormancy, the distribution of free IAA in tuber tissues by in situ analysis, and the biosynthesis of the hormone by feeding experiments. The time course of free IAA showed marked differences between the examined cultivars, although the concentration of the auxin generally was the highest at the early stages of tuber dormancy. Immunodetection showed a similar pattern of IAA distribution in both genotypes: in dormant buds from freshly harvested tubers, the free hormone accumulated mostly in apical meristem, leaf and lateral bud primordia, and differentiating vascular tissues underlying the apical meristem, while at the end of the storage period only axillary bud primordia from growing buds displayed appreciable auxin levels. Feeding experiments indicated that changes in IAA biosynthesis rate were a major cause of auxin variation in buds. In both cultivars, dormancy apparently ceased when free IAA fell below a threshold value. Despite this, our data led us to conclude that IAA would not be directly responsible for inhibiting sprouting. Instead, auxin might shorten dormancy, in a cultivar-dependent manner, by enhancing early developmental processes in buds, ultimately leading to dormancy termination.

The nucellus is a maternal tissue that embeds and feeds the developing embryo and secondary endosperm. During seed development, the cells of the nucellus suffer a degenerative process soon after fertilization as the cellular endosperm expands and accumulates reserves. Nucellar cell degeneration has been considered to be a form of developmentally programmed cell death (PCD). It was investigated whether or not this degenerative process is characterized by apoptotic hallmarks. Evidence showed that cell death is mostly localized in the border region of the tissue adjacent to the expanding endosperm. Cell death is accompanied by profound changes in the morphology of the nuclei and by a huge degradation of nuclear DNA. Moreover, an increase of activity of different classes of proteinases is reported, and the induction of caspase-like proteases sensitive to specific inhibitors was detected. Nucellar caspase-like proteases are characterized by an acid pH optimum suggesting a possible localization in the vacuole.

The suspensor of Phaseolus coccineus L. degenerates at the cotyledonary stage of embryogenesis when it is no longer necessary for continued embryonic development; this degeneration is considered to be a typical example of the so-called developmental programmed cell death (PCD) in plants. The presence of specific hallmarks of PCD as it occurs during the degeneration of P. coccineus suspensor was investigated in the current study. By using the TUNEL assay and electrophoretic analysis, we found evidence of nuclear DNA degradation, a known feature of PCD, in the endosperm and degenerating suspensors. Degeneration of the suspensor begins after degeneration in the endosperm and it starts in the neck region, spreading basipetally towards the knob. We conclude from this study that suspensor degeneration in P. coccineus occurs by means of PCD and displays typical hallmarks of PCD, such as DNA fragmentation. PCD in the suspensor is a highly asynchronous process, originating first in the neck cells and subsequently spreading to the basal cells.

Summary The intact fruit and isolated seed and fruit tissues of Sechium edule Swartz were analysed for ethylene production, ACC content and activity of the enzymes involved in ethylene biosynthesis (ACS and ACO) in consequence to wounding. Stress ethylene production follows different patterns according to which the different tissues of the fruit and seed are concerned. On the whole the data obtained indicate that the seed teguments are the only tissue that is characterised by an increase in ACS activity and in ethylene production in relation to stress.

Stomata respond to blue light and it is generally believed that the photoreceptor for this response is located inside the guard cells. Only a small number of blue light photoreceptors have been identified so far, namely cryptochromes and phototropins, and they show overlapping functions in regulating many different responses to light. The possibility that plants may possess other receptors regulating blue light responses under different light conditions cannot be excluded. In this paper we show the presence of two retinal-binding proteins in Commelina communis and we report the identification of retinal, a chromophore usually bound to the photoreceptor rhodopsin and previously identified in algae and other higher plants. We show that, under our experimental conditions, stomata open promptly when exposed to blue light and we demonstrated that this response is dependent on retinal. We hypothesise that rhodopsin-like retinal-binding proteins might be involved in stomatal response to blue light.

Immature embryos of Cytisus laburnum L. were cultivated in vitro and four culture media, different techniques of substrate preparation, sucrose concentration and the effect of suspensor removal were tested. The best results were obtained with N6 medium supplemented with 2 mg dm−3 glycine and set up using a double-layer culture system, in which the top layer had a higher osmotic potential than the bottom one. These conditions allowed normal embryogenic development in up to 45 % of early globular embryos, that were able to develop until a complete maturity. Osmotic potential and mineral nutrients of the medium demonstrated to be crucial for the successful culture and their effects were dependent on embryo age at the time of excision. The presence of an intact suspensor showed to be beneficial only for early globular embryos while older developmental stage embryos were not significantly affected.

In the present work, the characterization of two variant cell lines of carrot capable of growing in high (92 μmol L–1) concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) is reported. Both these cell lines (4w77 and 4w13) show a significantly lower uptake of 2,4-D with respect to wild-type (wt) cells. In contrast to wt cells, influx of IAA is not reduced by the addition of 100 μM 2,4-D and the presence of this compound appears to stimulate IAA uptake. When grown in the presence of high concentrations of 2,4-D, both 4w77 and 4w13 cells show behavioural differences: instead of lowering the endogenous level of free IAA, the two resistant lines react to the high exogenous concentrations of auxin by raising the level of the free hormone. In 4w77 cells, this is accomplished by reduction of auxin released in the external medium or converted to amide-linked conjugates. In 4w13 cells, the final level of endogenous IAA is an equilibrium between increased synthesis of IAA and a massive release into the medium of the ester- and free-forms of IAA. Both cell lines show disturbances in embryogenesis: line 4w77 forms globular embryos that only mature into aberrant forms having multiple axes, whereas line 4w13 has completely lost its morphogenic capacity.

In the present work we studied oxidative stress as an important cause of seed deterioration during ageing in embryos from durum wheat grains stored at room temperature and at low temperature (10°C). The protective role of low temperature on seed viability was confirmed. The increase of hydrogen peroxide content during dry storage was strongly correlated with the decrease of germinability. Ascorbate and glutathione showed a good correlation with grain germinability and significantly increased upon imbibition, in particular in embryos from viable grains. Ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX) and catalase (CAT) were studied quantitatively (enzymatic assays). APX, GR, and GPX were also studied qualitatively by native PAGE. The enzymes were active in dry, still viable, embryos whereas no activity was detected in non-viable embryos. With the exception of APX, all enzymatic activities decreased upon imbibition. The study of grains stored in different conditions indicated a negative correlation between the efficiency of the antioxidant enzymatic machinery and the age of the grain. The differences detected in differently stored materials confirmed that both germination parameters and the length of storage period are important in determining grain condition.

Summary To improve knowledge of the processes involved in fruit abscission, the abscisic acid levels and the structure of both whole seed and single seed tissues were investigated in control (sample A) and shedding affected fruits (sample B) of Malus domestica . ABA content was measured in crude extracts by radioimmunoassay and the results were validated by physicochemical methods. Whole seeds were embedded and sectioned for light and electron microscopy studies. The ABA level in whole seeds was almost three times higher in the affected fruits than in the control. No difference in ABA content was observed between the embryos of the two samples, while in the endosperm, the nucellus and especially in the integuments, the ABA level was clearly higher in sample B than in sample A. Delayed development and precocious cell differentiation as well as certain aspects of whole system degeneration were observed. The authors conclude that ABA levels seem to be one of the factors involved in fruit abscission in an early period of development.

Biosynthesis of gibberellins (GAs) was studied in vivo in endosperms of Sechium edule Sw. Exogenous ent-[(14)C]kaurene was metabolized into four major products: GA(12), GA(4), GA(7) and 16, 17-dihydro-16-hydroxy-GA(15) alcohol glucoside. Other minor metabolites were also observed including ent-kaurenol and ent-kaurenal. Conversion of ent-[(14)C]kaurene to ent-kaurenol glucoside by endosperm cell-free preparations in the presence of UDPG was observed. However, the finding was not confirmed in in vivo studies and is probably artifactual. Overall evidence coming from the analysis of endogenous GAs and in vitro and in vivo biosynthetic studies are discussed in relation to the possible existence in the Sechium seeds of a different route, along with the known pathway, branching from ent-kaurene or ent-7-alpha-hydroxykaurenoic acid and this also leading to biologically active GAs.

We here report the purification and identification of all- trans -retinal by column chromatography, HPLC and GC-MS in a tomato mutant ( Lycopersion esculentum Mill. aurea mutant). Since retinal, wherever present, is almost exclusively associated to an opsin to form a light-sensitive complex (rhodopsin-like proteins), this finding suggests that a rhodopsin-like protein could function as a blue light photoreceptor in higher plants.

The time course of free and conjugated indole-3-acetic acid (IAA) concentrations was investigated in three distinct parts (eyes, subeye tissues and pith) of potato (Solanum tuberosum L. cultivar Monalisa) tubers, from the last period of tuber growth to sprouting under two different storage temperatures (3 and 23˚C). The highest levels of both free and conjugated IAA were detected in eyes, which showed a significant increase of the free hormone concentration from harvest to the end of dormancy, regardless of storage temperature. The concentration of conjugates was higher than free IAA in eyes and subeye tissues at 23˚C, while at 3˚C conjugates prevailed only in eyes. Present results suggest the involvement of IAA in the control of potato tuber dormancy.

(1995). Molecular and Biochemical Characterization of 2.4D Resistant Carrot Cell Lines. Giornale botanico italiano: Vol. 129, No. 4, pp. 996-996.

Summary Apple fruits were collected in the period between 20 and 49 days after anthesis. The relation between histological structure and gibberellin content was investigated in ovules of control fruits (sample A), fruitdrop affected fruits not yet abscinded from the plant (sample B) and fruit-drop affected fruits already abscinded from the plant (sample C). The results of the histological analyses showed that: i) ovules from sample A generally contained a developing embryo, a dividing endosperm, and a conspicuous antipodal tissue; ii) ovules from sample B were degenerating. However, they contained a degraded embryo mainly in fruits collected 29 days post anthesis. No endosperm or antipodal cells were detected; iii) ovules from samples C also degenerated and had no recognizable structure. Gibberellins were determined in ovules isolated from the same three samples of fruits. The gibberellin content was very low in all samples and quantitative differences between sample A, on the one hand, and samples Band C on the other, were only detectable 49 days after anthesis. Moreover, biological activity appeared to be distributed in different Rf-zones in the TL chromatogram, suggesting the presence of various gibberellin-like substances. The differences observed were confirmed after HPLC purification. Further analyses by means of GC and GC - MS led to the identification of GA4 and GA63 as two of the GAs present in ovules from sample A. The results obtained are discussed in relation to fruit abscission heterogeneity.

Growth response of coleoptile sections from four Italian cultivars of wheat to some growth regulators.—The following Italian wheat varieties were used to obtain the coleoptile sections used throughout the experiments: Funo, S. Pastore, Abbondanza, Generoso. The growth substances tested were Indolacetic acid (IAA), Abscisic acid (ABA), Kinetin and Gibberellic acid. The colcoptile response of each variety was tested with three different concentrations for each growth regulator in combination with two sucrose concentrations and three different pH levels. The experiments indicated that the coleoptiles from the variety « Funo » react linearly to incresing IAA and ABA concentration resulting the best one to be used in this bioassay.

Endosperm and cotyledons of Sechium edule at different stages of seed development were found to contain three novel GA conjugates. 3-propyl- or 3-acetyl- GA 4/7 were characterized by mass spectral analysis and were found to be biologically active in contrast to synthetic GA 4/7 propyl esters. An unusual GA glucoside showing biological activity was also isolated from both tissues and characterized by mass spectrometry and NMR analysis as 16,17-dihydro-16-hydroxy-GA 15 alcohol glucoside. The functional significance of this novel GA conjugate is discussed.

Summary Seeds of higher plants are known to be rich in cytokinins. However the single seed components have been infrequently isolated and analysed, studies being usually referred to the whole seed. Data are presented on the occurrence and identity of cytokinins in the endosperm of Sechium edule seeds at different developmental stages. The results demonstrate the presence of 2iPNT, ZNT, ZR, Z, ZOG and ZROG. The possible significance of cytokinin nucleotides, ribosides, free bases and O-glucosides in this tissue is discussed.

Auxin contents have been evaluated at different times during the rooting period in two rose rootstocks with a different rooting capacity. Avena first internode has been used as assay. The activity of the acidic phase of the extracts decreases in both species but more so in the easy-to-root species Rosa multiflora. The lowest activity of the extract occurs at the 14th day; whereas at the 21st day, the activity starts to increase again. The activity of the basic phase decreases in the same way in both varieties but at the 14th day in Rosa multiflora it increases again. Rooting failure is more probably due to some step of the reactions which lead to root formation being inhibited rather than to auxin deficiency.

Cell-free extracts prepared from endosperms and cotyledons of Sechium edule seeds at four stages of growth were incubated with [2- 14 C]mevalonic acid to investigate their capacity for ent -kaurene. Both tissues synthesize ent -kaurene at each stage of seed growth. The rate of synthesis was higher in cotyledons than in endosperms and was the same at each stage examined. Endosperm and cotyledon cell-free extracts prepared from seeds at the same stage of growth were incubated with 14 C-labelled ent -kaurene. In the absence of Fe 2+ , dialysed extracts of both tissues metabolized ent -kaurene to ent -kaurenol, ent -kaurenal, ent -kaurenoic acid, ent -7 β-hydroxykaurenoic acid and GA 12 -aldehyde. In the presence of Fe 2+ , non-dialysed extracts were able to synthesize gibberellins. The radioactivity from ent -kaurene was incorporated in to the same products that accumulate endogenously both in endosperms and cotyledons, i.e. GA 4 and/or GA 7 and GA 9 . Radioactive GA 15 was also detected in the incubation mixtures.

Gibberellins (GAs) in suspensors and embryos of Phaseolus coccineus seeds at the heart stage of embryo development were analyzed by combined gas chromatography-mass spectrometry (GC-MS). From the suspensor four C(19)-GAs, GA(1), GA(4), GA(5), GA(6), and one C(20) GA, GA(44), were identified. From the embryo, five C(19)-GAs GA(1), GA(4), GA(5), GA(6), GA(60) and two C(20) GAs, GA(19) and GA(44) were identified. The data, in relation to previous results, suggest a dependence of the embryo on the suspensor during early stages of development.

The time-course growth of single tissues in pollinated and unpollinated ovules of Sechium edule Sw. is described in relation to the endogenous levels of abscisic acid. Quantitation of abscisic acid (ABA) in the minute amounts of material obtained after ovule dissection has been performed by using a highly specific and sensitive solid-phase radioimmunoassay based on a monoclonal antibody raised against free (S)-ABA. While the absolute amount of ABA rises in both types of ovules, only in unpollinated ones does this leads to an increase in the hormone concentration. Infact in pollinated ovules the rapid growth following pollination prevents, through a dilution effect, the increase in ABA concentration. Growth patterns and endogenous ABA levels are similar for integuments and nucellus tissues either in pollinated or unpollinated ovules. It is suggested that the growth inhibition induced by the increase in ABA concentration after anthesis could be counteracted by the pollination triggered fast ovule growth.

Summary A new mouse monoclonal antibody against abscisic acid (ABA) was produced and characterized. This antibody is highly specific for free (S)-ABA and shows a good affinity constant. A newly developed solid phase radioimmunoassay (RIA) for the quantitation of ABA content in plant extracts is described. This assay proved to be more sensitive and precise than the commonly used liquid phase RIA. Using this assay ABA was quantified in tomato and bean leaves and in Sechium endosperm. Results were compared with those obtained using the other two monoclonal antibodies produced up to now against ABA. Results obtained with RIA were validated by physicochemical methods.

A polar gibberellin-like substance present in needles of Picea sitchensis was identified as GA 9 -β- d -glucosyl ester on the basis of enzymatic hydrolysis and identification of the aglycone by GC-MS. The biological activity of the synthetic material was tested in two bioassays.