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La presente investigación analiza la faceta coleccionista de la reina Bárbara de Braganza tomando como referencia a su predecesora en el trono español, Isabel de Farnesio, y a partir de las pinturas y piezas de orfebrería que donó al monasterio de las Salesas Reales. Para ello analizamos previamente la educación recibida y su papel en la corte de Fernando VI. Adjuntamos la transcripción del inventario correspondiente a las pinturas; Esta pesquisa analisa o lado do colector da rainha Bárbara de Bragança, com referência ao seu antecessor no trono espanhol, Isabel de Farnesio, e de pinturas e peças de ourives doadas ao mosteiro da Salesas Reales. Para isso, analisou recebeu previamente educação e seu papel na corte de Fernando VI. A transcrição do correspondente ligado ao inventário pinturas.; The present investigation analyzes the side collector of the queen Bárbara de Braganza taking as reference to its predecessor in the Spanish throne, Isabel de Farnesio, and from the paintings and goldsmithing pieces that donated to the monastery of the Salesas Reales. For this we analyze previously the education received and its paper in the court of Fernando VI. Attached is the transcript of the inventory corresponding to the paintings

Arbuscular mycorrhizal (AM) symbioses are mutualistic associations between soil fungi and most vascular plants. Their association benefits the host plant by improving nutrition, mainly phosphorus nutrition, and by providing increased capability to cope with adverse conditions. In this study, we investigated the transcriptional changes triggered in rice leaves as a result of AM symbiosis, focusing on the relevance of the plant defence response. We showed that root colonization by the AM fungus Glomus intraradices is accompanied by the systemic induction of genes that play a regulatory role in the host defence response, such as OsNPR1, OsAP2, OsEREBP and OsJAmyb. Genes involved in signal transduction processes (OsDUF26 and OsMPK6) and genes that function in calcium-mediated signalling processes (OsCBP, OsCaM and OsCML4) are also up-regulated in leaves of mycorrhizal rice plants in the absence of pathogen infection. In addition, the mycorrhizal rice plants exhibit a stronger induction of defence marker genes [i.e. pathogenesis-related (PR) genes] in their leaves in response to infection by the blast fungus Magnaporthe oryzae. Evidence indicates that mycorrhizal rice plants show enhanced resistance to the rice blast fungus. Overall, these results suggest that the protective effect of the AM symbiosis in rice plants relies on both the systemic activation of defence regulatory genes in the absence of pathogen challenge and the priming for stronger expression of defence effector genes during pathogen infection. The possible mechanisms involved in the mycorrhiza-induced resistance to M. oryzae infection are discussed.

Gibberellins are phytohormones that regulate growth and development of plants. Gibberellin homeostasis is maintained by feedback regulation of gibberellin metabolism genes. To understand this regulation, we manipulated the gibberellin pathway in tobacco and studied its effects on the morphological phenotype, gibberellin levels and the expression of endogenous gibberellin metabolism genes. The overexpression of a gibberellin 3-oxidase (biosynthesis gene) in tobacco (3ox-OE) induced slight variations in phenotype and active GA(1) levels, but we also found an increase in GA(8) levels (GA(1) inactivation product) and a conspicuous induction of gibberellin 2-oxidases (catabolism genes; NtGA2ox3 and -5), suggesting an important role for these particular genes in the control of gibberellin homeostasis. The effect of simultaneous overexpression of two biosynthesis genes, a gibberellin 3-oxidase and a gibberellin 20-oxidase (20ox/3ox-OE), on phenotype and gibberellin content suggests that gibberellin 3-oxidases are non-limiting enzymes in tobacco, even in a 20ox-OE background. Moreover, the expression analysis of gibberellin metabolism genes in transgenic plants (3ox-OE, 20ox-OE and hybrid 3ox/20ox-OE), and in response to application of different GA1 concentrations, showed genes with different gibberellin sensitivity. Gibberellin biosynthesis genes (NtGA20ox1 and NtGA3ox1) are negatively feedback regulated mainly by high gibberellin levels. In contrast, gibberellin catabolism genes which are subject to positive feedback regulation are sensitive to high (NtGA2ox1) or to low (NtGA2ox3 and -5) gibberellin concentrations. These two last GA2ox genes seem to play a predominant role in gibberellin homeostasis under mild gibberellin variations, but not under large gibberellin changes, where the biosynthesis genes GA20ox and GA3ox may be more important.

El presente artículo analiza la configuración de la colección de arte de los condes de Parcent, centrándonos en aquellos bienes artísticos cuya presencia es más significativa en los inventarios de la Casa: pinturas, tapices, joyas y plata. Para analizar esta colección, nos hemos centrado en el origen italiano de este linaje, su parentesco con el papa Inocencio XI y su actividad como prestamistas, como hechos diferenciadores de su faceta coleccionista.A lo que se añade la incorporación a la Casa, del condado de Contamina y el marquesado de Fuente el Sol, incorporando importantes conjuntos artísticos, tanto muebles como inmuebles.; Questo articolo analizza la configurazione della collezione d’arte dei conti di Parcent, concentrandosi su quei beni artistici la cui presenza è più significativa negli inventari della Casa: dipinti, arazzi, gioielli e d’argento. Per analizzare questa collezione, ci siamo concentrati sulla origine italiana di questo lignaggio, il suo rapporto con Papa Innocenzo XI e la sua attività di istituti di credito, come differenziare i fatti del suo lato collettore. A cui si aggiunge la costituzione della Casa, nella contea di Contamina e Marchese di Fuente el Sol, integrando importanti formazioni artistiche sia mobili e immobili; This article analyzes the configuration of the art collection of the counts of Parcent, focusing on those artistic assets whose presence is more significant in the inventories of the House: paintings, tapestries, jewelry and silver. To analyze this collection, we have focused on the Italian origin of this lineage, his relationship with Pope Innocent XI and his activity as lenders, as differentiating facts of his collector side. To which the incorporation of the House, County Contamina and Marquis of Fuente el Sol is added, incorporating important artistic ensembles both movable and immovable.

The role of the apical shoot as a source of inhibitors preventing fruit growth in the absence of a stimulus (e.g. pollination or application of gibberellic acid) has been investigated in pea (Pisum sativum L.). Plant decapitation stimulated parthenocarpic growth, even in derooted plants, and this effect was counteracted by the application of indole acetic acid (IAA) or abscisic acid (ABA) in agar blocks to the severed stump. The treatment of unpollinated ovaries with gibberellic acid blocked the effect of IAA or ABA applied to the stump. [3H]IAA and [3H]ABA applied to the stump were transported basipetally, and [3H]ABA but not [3H]IAA was also detected in unpollinated ovaries. The concentration of ABA in unpollinated ovaries increased significantly in the absence of a promotive stimulus. The application of IAA to the stump enhanced by 2- to 5-fold the concentration of ABA in the inhibited ovary, whereas the inhibition of IAA transport from the apical shoot by triiodobenzoic acid decreased the ovary content of ABA (to approximately one-half). Triiodobenzoic acid alone, however, was unable to stimulate ovary growth. Thus, in addition to removing IAA transport from the apical shoot, the accumulation of a promotive factor is also necessary to induce parthenocarpic growth in decapitated plants.

Conselleria Agr Pesca & Alimentac Generalitat Valenciana; Min Flemish Commun, Adm Agr & Horticulture Belgium

Using two lowland rice (Oryza sativa L.) cultivars we found that in both cases submerged-induced elongation early after germination depends on gibberellins (GAs). Submergence increases the content of the active GA1 by enhancing the expression of GA biosynthesis genes, thus facilitating the seedlings to escape from the water and preventing asphyxiation. However, the two cultivars differ in their response to ethylene. The cultivar Senia (short), by contrast to cultivar Bomba (tall), does not elongate after ethylene application, and submerged-induced elongation is not negated by an inhibitor of ethylene perception. Also, while ethylene emanation in Senia is not altered by submergence, Bomba seedlings emanate more ethylene upon de-submergence, associated with enhanced expression of the ethylene biosynthesis gene OsACS5. The cultivar Senia thus allows the possibility of clarifying the role of ethylene and other factors as triggers of GA biosynthesis enhancement in rice seedlings under submergence.

[EN] We examined the gibberellin (GA) and ethylene regulation of submergence-induced elongation in seedlings of the submergence-tolerant lowland rice (Oryza sativa L) cvs Senia and Bomba. Elongation was enhanced after germination to facilitate water escape and reach air. We found that submergence-induced elongation depends on GA because it was counteracted by paclobutrazol (an inhibitor of GA biosynthesis), an effect that was negated by GA(3). Moreover, in the cv Senia, submergence increased the content of active GA(1) and its immediate precursors (GA(53), GA(19) and GA(20)) by enhancing expression of several GA biosynthesis genes (OsGA20ox1 and -2, and OsGA3ox2), but not by decreasing expression of several OsGA2ox (GA inactivating genes). Senia seedlings, in contrast to Bomba seedlings, did not elongate in response to ethylene or 1-aminocyclopropane-1-carboxylic-acid (ACC; an ethylene precursor) application, and submergence-induced elongation was not reduced in the presence of 1-methylcyclopropene (1-MCP; an ethylene perception inhibitor). Ethylene emanation was similar in Senia seedlings grown in air and in submerged-grown seedlings following de-submergence, while it increased in Bomba. The expression of ethylene biosynthesis genes (OsACS1, -2 and -3, and OsACO1) was not affected in Senia, but expression of OsACS5 was rapidly enhanced in Bomba upon submergence. Our results support the conclusion that submergence elongation enhancement of lowland rice is due to alteration of GA metabolism leading to an increase in active GA (GA1) content. Interestingly, in the cv Senia, in contrast to cv Bomba, this was triggered through an ethylene-independent mechanism. Published by Elsevier GmbH, We thank D. Luis Marques (Copsemar, Sueca, Valencia, Spain) for the generous gift of Senia and Bomba seeds, Floralife (Walterboro, SC, USA) for supply of 1-MCP, Monsanto (St. Louis, Missouri, USA) for providing sequence information for rice BACs, and Ms. Teresa Sabater for technical help with the experiments. The work was supported by a European Union Research Training Network (RTN1-1999-00086) and BIO2009-07968.

CcGA20ox1 is a gene encoding a GA 20-oxidase, a gibberellin (GA) biosynthetic enzyme, previously isolated from the citrus hybrid Carrizo citrange (Citrus sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.). Southern blot analysis of genomic DNA of Carrizo citrange with CcGA20ox1 suggested the presence in the hybrid of another gene encoding another GA 20-oxidase. A cDNA clone from this new gene (CcGA20ox2) was isolated using RNA from the other parent C. sinensis. CcGA20ox2 encoded a protein of 372 amino acids that showed 67.1% identify with CcGA20ox1, and its expression product catalyzed the in vitro conversion of GA(12) to GA(9), confirming that it corresponds to another active GA20ox. Amplification of genomic DNA and isolation of genomic clones of CcGA20ox1 and CcGA20ox2 revealed that the parental sources these genes in the hybrid were P. trifolicata and C. sinensis, respectively. The sequences of CcGA20ox1 and CcGA20ox2 showed that both genes contained two introns, which are also conserved in GA20ox genes of other species like Arabidopsis thaliana L., Pisum sativum L. and Solanum lycopersicum L. Determination of transcript levels in the Carrizo citrange hybrid by quantitative real-time polymerase chain reaction showed that CcGA20ox1 and expressed mainly in internodes, leaves and seeds, and CcGA20ox2 in flower buds and flowers at anthesis, with the genes having similar transcript levels in young developing fruits.

Flowering of Nicotiana tabacum cv Xhanti depends on gibberellins because gibberellin-deficient plants, due to overexpression of a gibberellin 2-oxidase gene (35S.NoGA2ox3) or to treatment with the gibberellin biosynthesis inhibitor paclobutrazol, flowered later than wild type. These plants also showed inhibition of the expression of molecular markers related to floral transition (NtMADS-4 and NtMADS-11). To investigate further the role of gibberellin in flowering, we quantified its content in tobacco plants during development. We found a progressive reduction in the levels of GA(1) and GA(4) in the apical shoot during vegetative growth, reaching very low levels at floral transition and beyond. This excludes these two gibberellins as flowering-promoting factors in the apex. The evolution of active gibberellin content in apical shoots agrees with the expression patterns of gibberellin metabolism genes: two encoding gibberellin 20-oxidases (NtGA20ox1=Ntc12, NtGA20ox2=Ntc16), one encoding a gibberellin 3-oxidase (NtGA3ox1 = Nty) and one encoding a gibberellin 2-oxidase (NtGA2ox1), suggesting that active gibberellins are locally synthesized. In young apical leaves, GA(1) and GA(4) content and the expression of gibberellin metabolism genes were rather constant. Our results support that floral transition in tobacco, in contrast to that in Arabidopsis, is not regulated by the levels of GA(1) and GA(4) in apical shoots, although reaching a threshold in gibberellin levels may be necessary to allow meristem competence for flowering.

Carrizo citrange (Citrus sinensis x Poncirus trifoliata) is a citrus hybrid widely used as a rootstock, whose genetic manipulation to improve different growth characteristics is of high agronomic interest. In this work, transgenic Carrizo citrange plants have been produced overexpressing sense and antisense CcGA20ox1 (a key enzyme of GA biosynthesis) under control of the 35S promoter to modify plant architecture. As expected, taller (sense) and shorter (antisense) phenotypes correlated with higher and lower levels, respectively, of active GA(1) in growing shoots. In contrast, other phenotypic characteristics seemed to be specific to citrus, or different from those described for similar transgenics in other species. For instance, thorns, typical organs of citrus at juvenile stages, were much longer in sense and shorter in antisense plants, and xylem tissue was reduced in leaf and internode of sense plants. Antisense plants presented a bushy phenotype, suggesting a possible effect of GAs on auxin biosynthesis and/or transport. The main foliole of sense plants was longer, although total leaf area was reduced. Leaf thickness was smaller in sense and larger in antisense plants due to changes in the spongy parenchyma. Internode cell length was not altered in transgenic plants, indicating that, in citrus, GAs regulate cell division rather than cell elongation. Interestingly, the phenotypes described were not apparent when transgenic plants were grafted on non-transgenic rootstock. This suggests that roots contribute to the GA economy of aerial parts in citrus and opens the possibility of using the antisense plants as dwarfing rootstocks.

CLADOSPORIUM-FULVUM; DISEASE RESISTANCE; BRASSINOSTEROID BIOSYNTHESIS; GIBBERELLIN CONTENT; BETA-GLUCURONIDASE; ARABIDOPSIS; EXPRESSION; PROTEINS; PLANTS; GROWTH

A functional genomics project has been initiated to approach the molecular characterization of the main biological and agronomical traits of citrus. As a key part of this project, a citrus EST collection has been generated from 25 cDNA libraries covering different tissues, developmental stages and stress conditions. The collection includes a total of 22,635 high-quality ESTs, grouped in 11,836 putative unigenes, which represent at least one third of the estimated number of genes in the citrus genome. Functional annotation of unigenes which have Arabidopsis orthologues (68% of all unigenes) revealed gene representation in every major functional category, suggesting that a genome-wide EST collection was obtained. A Citrus clementina Hort. ex Tan. cv. Clemenules genomic library, that will contribute to further characterization of relevant genes, has also been constructed. To initiate the analysis of citrus transcriptome, we have developed a cDNA microarray containing 12,672 probes corresponding to 6875 putative unigenes of the collection. Technical characterization of the microarray showed high intra- and inter-array reproducibility, as well as a good range of sensitivity. We have also validated gene expression data achieved with this microarray through an independent technique such as RNA gel blot analysis.

Plants undergo two different developmental programs depending on whether they are growing in darkness (skotomorphogenesis) or in the presence of light (photomorphogenesis). It has been proposed that the latter is the default pathway followed by many plants after germination and before the seedling emerges from soil. The transition between the two pathways is tightly regulated. The conserved COPI-based complex is central in the light-dependent repression of photomorphogenesis in darkness. Besides this control, hormones such as brassinosteroids (BRs), cytokinins, auxins, or ethylene also have been shown to regulate, to different extents, this developmental switch. In the present work, we show that the hormone gibberellin (GA) widely participates in this regulation. Studies from Arabidopsis show that both chemical and genetic reductions of endogenous GA levels partially derepress photomorphogenesis in darkness. This is based both on morphological phenotypes, such as hypocotyl elongation and hook and cotyledon opening, and on molecular phenotypes, such as misregulation of the light-controlled genes CAB2 and RNS. Genetic studies indicate that the GA signaling elements GAI and RGA participate in these responses. Our results also suggest that GA regulation of this response partially depends on BRs. This regulation seems to be conserved across species because lowering endogenous GA levels in pea (Pisum sativum) induces full de-etiolation in darkness, which is not reverted by BR application. Our results, therefore, attribute an important role for GAs in the establishment of etiolated growth and in repression of photomorphogenesis.

Fruit-set and fruit growth in pea (Pisum sativum L.) depend on gibberellins (GAs). The authors have isolated a new pea mutant, gio, which appeared spontaneously within the population of the cultivar Alaska, characterized by unpollinated ovaries much less sensitive to applied GAs. The mutant also has elongated peduncles, and is taller than the wild-type (WT) because the upper plant internodes are longer. Contrary to WT, the gio ovaries respond very little to benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid, but become fully sensitive to GA(3) when this hormone is applied together with BAP. The gio phenotype is determined by a mutation at a single mendelian locus. The mutation is recesive, shows incomplete penetrance, and its expression depends on environmental culture conditions. The sensitivity of the ovaries to GA(3) can be recovered by removing the apical shoot (plant decapitation) and by blocking the transport of indoleacetic acid (IAA) from the apical shoot with 2,3,5-triiodobenzoic acid. The content of IAA in methanolic extracts and phloematic exudates of the apical shoot of gio is about double that in the WT. The rate of transport of [(3)H]IAA applied to the apex of the mutant is also twice that in the WT. This indicates that the insensitivity of the gio ovaries to GAs is due to the inhibitory effect of the higher basipetal IAA transport from the shoot. The interaction between the fruit and the apical shoot mediated by IAA probably also involves cytokinins transported from the basal part of the plant.

The regulation by phytochrome of stem elongation in light-grown plants depends on gibberellins (GAs). To investigate whether this is mediated by a change in GA metabolism, the effect of the GA biosynthesis inhibitor LAB 198 999 (an acylcyclohexadione derivative) on the end-of-day far-red (FR) response in cowpea (Vigna sinensis L.) epicotyl explants has been investigated. Growth of epicotyl explants of light-grown seedlings was enhanced when treated with far-red light before incubation in the dark (end-of-day PR effect). Low doses of LAB 198 999 (0.05 and 0.5 mu g explant(-1)) reduced the effect of FR, whereas 5 to 50 mu g explant-l stimulated elongation of both red light (R)- and FR-treated epicotyl explants while nullifying the differences between R and FR treatments. In paclobutrazol-treated epicotyl explants, FR enhanced the response to applied GA(1) and GA(20), whereas LAB 198 999 increased the activity of GA(1) and decreased that of GA(20). [H-3]Gibberellin A(1), injected into the basal Dart of the epicotyl, was transported and metabolized mainly to [H-3]GA(8) in the apical 20 mm of the epicotyl. The conversion of [H-3]GA(1) to [H-3]GA(8) was dramatically reduced by both end-of-day FR treatments and LAB 198 999 applications. In addition, both treatments enhanced epicotyl elongation. It is proposed that the regulation of cowpea epicotyl growth by phytocrome is mediated, at least partially, by modifying GA(1) degradation.

There is evidence suggesting that gibberellins (GAs) may be involved in the modulation of stem elongation by light (Reid, 1983), though the nature of this interaction is not clear. The elongation of the cowpea epicotyl is controlled by GAs (Garcia-Martinez and Rappaport, 1984), and it is modulated by phytochrome (Garcia-Martinez et al. 1987). It has been suggested that phytochrome regulates GA20 metabolism in cowpea epicotyl (Garcia-Martinez et al. 1987). In this communication we report on the effect of red (R) and far red (FR) irradiation on the metabolism of [3H]GA20 and [3H]GAi. Phytochrome seems to affect the metabolism of both GAs.

The effect of LAB 198 999 [3,5-dioxo-4-butyrylcyclohexane carboxylic acid ethyl ester; a new plant growth retardant which competitively inhibits 2-oxoglutarate-dependent gibberellin (GA) dioxygenases] on elongation and in-vivo [H-3]GA1 and [H-3]GA20 metabolism in cowpea (Vigna sinensis L. cv Blackeye pea No. 5) epicotyls has been investigated. Gibberellins and LAB 198 999 were injected into the epicotyl at 25-30 mm from the apex. In intact seedlings, epicotyl elongation was inhibited by LAB 198 999 (25-mu-g . epicotyl-1), and the inhibition was counteracted by GA1 but not by GA20. In contrast to intact seedlings, the inhibitor enhanced epicotyl elongation in de-bladed seedlings and explants, in the latter case proportionally to the amount of inhibitor applied (up to 50-mu-g . epicotyl-1), but not in explants made from paclobutrazol-treated seedlings. The inhibitor also enhanced dramatically the elongation induced in paclobutrazol-treated explants by GA1, but not by GA20. The promotive effect of LAB 198999 was associated with increased contents of GA1 and GA8 in the growing region of the epicotyl, indicating a dependence on endogenous GAs. The effect of LAB 198 999 decreased progressively with the age of the seedlings, probably as a consequence of a decreased level of GAs in the epicotyl. Gibberellin substrates and metabolites in the growing region of the epicotyl (upper 20 mm) were fractionated and identified tentatively by high-performance liquid chromatography and radiocounting using a homogeneous on-line radioactivity detector. The metabolism of [H-3]GA1(t) (tentative) to [H-3]GA8(t), and that of [H-3]GA20(t) to [H-3]GA1(t) and [H-3]GA29(t) in the epicotyl were blocked by LAB 198 999, that of the former more efficiently than the latter. The results presented support the hypothesis that GA1 is the active GA controlling elongation of cowpea epicotyls. They also show that both the promotion of epicotyl elongation in explants and the enhancement of the effect of exogenous GA, by LAB 198 999 are the result of the inhibitor blocking the in-vivo 2-beta-hydroxylation of GA1.

The histological development of fertilized ovules during fruit-set and development in pea (Pisum sativum L. cv. Alaska) has been investigated. Killing the ovules on day 0 (anthesis) or day 1 prevented fruit-set and resulted in ovary degeneration. When the ovules were destroyed at later stages the ovaries developed, though the rate of growth of the pod was reduced significantly. Pollination in pea occurs normally the day before anthesis, and fertilization of the egg cell 32 to 48 h later. The first divisions of the zygote and endosperm nuclei started simultaneously (ca 48 h after pollination) but the endosperm developed more rapidly than the embryo; the embryo sac cavity was lined with free endosperm nuclei at the time of beginning suspensor elongation. Extracts of endosperm and ovule coats from ovules at day 7 after anthesis showed fruit-set activity in pea, the latter material having about 3 times more activity than the former per ovule basis. These results indicate that fertilization of the ovule is necessary for fruit-set in pea, and that compounds which induce fruit-set are probably synthesized in the ovules following fertilization.

S. tuberosum ssp. andigena potato plants require short days (SD) for tuberization. We have isolated PHOR1 (photoperiod-responsive 1), which shows upregulated expression in induced leaves (SD). PHOR1 encodes an arm repeat protein with homology to the Drosophila segment polarity protein armadillo. Antisense inhibition of PHOR1 produces a semidwarf phenotype similar to that of GA-deficient plants, and the antisense lines show reduced GA responsiveness combined with a higher endogenous GA content than wild-type plants. Feedback regulation of GA biosynthetic genes is also altered in these lines. Conversely, transgenic lines overexpressing PHOR1 show an enhanced response to GA. GA application induces rapid migration of PHOR1-GFP protein to the nucleus. Thus, PHOR1 appears to be a general component of GA signaling pathways that relocalizes to the nucleus in the presence of GA.

Samples of tracheal sap of Citrus sinensis (L.) Osbeck cv. Washington Navel were taken from field trees throughout the year and the nitrogen composition of the sap was determined. The nitrogenous fraction of the sap was composed mainly of free amino acids (92–97% of total nitrogen) and nitrates throughout the year. Proline was the most abundant amino acid during almost the entire cycle, and its concentration was especially high during the autumn and winter period. Nevertheless, a significant part (40–60%) of the total organic nitrogen was transported as arginine. Total nitrogen as well as amino acids and nitrates were maximal at spring flush. At spring flush and summer flush there was also a diversification of α-amino nitrogen among different amino acids. During the spring flush, nitrates, asparagine and γ-aminobutyric acid in the xylem sap seemed to have a radicular origin, whereas glutamic acid and arginine were released from the surrounding parenchyma. The results suggest a metabolic transformation in the wood parenchyma of nitrogenous compounds coming from the roots (including reduction of nitrates) and a turnover of different nitrogen metabolites between the xylem and surrounding cells.

The concentration of some plant growth substances was studied in healthy and citrus exocortis viroid (CEV)-infected Gynura aurantiaca DC plants. No changes in the levels of abscisic acid were observed. As a result of infection by the viroid, an auxin-like substance of unknown chemical nature was formed, whereas the concentrations of auxin-like substance(s) with the same chromatographic properties as indolacetic acid remained unchanged. A significant decrease in the levels of endogenous gibberellins (probably GA3 and/or GA1) was observed in CEV-infected plants. This decrease may have been associated with the stunting exhibited by infected plants. Application of exogenous GA3 increased the relative infectivity index of CEV in G. aurantiaca when applied starting at 7 days after inoculation.

The polypeptide patterns obtained by sodium dodecylsulphate-polyacrylamide gel electrophoresis of undigested and autodigested extracts from pea (Pisum sativum L.) ovaries at the early stages of development or degeneration have been studied. Development of unpollinated ovaries was stimulated by application of different plant growth regulators (gibberellic acid, 2,4-dichlorophenoxyacetic acid, and N(6)-benzyladenine) or by plant topping. Polypeptide bands of similar mobility to ribulose-1,5-bisphosphate carboxylase (RuBPCase) subunits (16 and 55 kDa) could be detected in all types of autodigested extracts from stimulated ovaries. However these bands were absent in electrophoretic patterns of autodigested extracts from unstimulated ovaries after 3 d post anthesis and in patterns of autodigested mixtures of these extracts with either those from stimulated ovaries or those from unstimulated ovaries before day 3. These observations indicate that a proteolytic activity which promotes the hydrolysis of RuBPCase appears in unstimulated ovaries about 3 d after anthesis. This event coincides with the loss of the capacity of unpollinated ovaries to develop in response to gibberellic acid and with the degeneration of the ovary wall.

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBP carboxylase, EC 4.1.1.39) has been purified from orange [Citrus sinensis (L.) Osbeck cv. Washington Navel] leaves using sucrose gradient centrifugation in a fixed angle rotor. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two major bands corresponding to the two subunits of RuBP carboxylase were found. The large subunit coincided with the polypeptide band that has been previously reported to be preferentially mobilized during the spring and summer flush periods. The degradation of RuBP carboxylase during autodigestion of Citrus leaf extracts, investigated by SDS-PAGE, occurred mainly at acidic (2.5-5.5) pH. The two subunits showed differences in the rate of degradation, the smaller being more rapidly hydrolyzed than the larger. At least four proteolytic activities were identified by means of inhibitor experiments: 1) a pepstatin A-sensitive activity that acts on both RuBP carboxylase subunits, 2) a mercurial (p-hydroxymercuribenzoate and p-chloromercuriphenylsulfonate)-sensitive activity that degrades only the small subunit, 3) an EDTA-sensitive activity that hydrolyzes both the large and small subunits, and 4) a mercurial-stimulated activity that acts only on the large subunit. It is suggested that the last two proteases may be responsible for the degradation of RuBP carboxylase observed in vivo during the periods of mobilization of leaf protein in Citrus.

The role and source of gibberellins (GAs) involved in the development of parthenocarpic fruits of Pisum sativum L. has been investigated. Gibberellins applied to the leaf adjacent to an emasculated ovary induced parthenocarpic fruit development on intact plants. The application of gibberellic acid (GA3) had to be done within 1 d of anthesis to be fully effective and the response was concentration-dependent. Gibberellin A1 and GA3 worked equally well and GA20 was less efficient. [(3)H]Gibberellin A1 applied to the leaf accumulated in the ovary and the accumulation was related to the growth response. These experiments show that GA applied to the leaf in high enough concentration is translocated to the ovary. Emasculated ovaries on decapitated pea plants develop without application of growth hormones. When [(3)H] GA1 was applied to the leaf adjacent to the ovary a substantial amount of radioactivity accumulated in the growing shoot of intact plants. In decapitated plants, however, this radioactivity was mainly found in the ovary. There it caused growth proportional to the accumulation of CA1. Application of LAB 150978, an inhibitor of GA biosynthesis, to decapitated plants inhibited parthenocarpic fruit development and this inhibition was counteracted by the application of GA3 (either to the fruit, or the leaf adjacent to the ovary, or through the lower cut end of the stem). All evidence taken together supports the view that parthenocarpic pea fruit development on topped plants depends on the import of gibberellins or their precursors, probably from the vegetative aerial parts of the plant.

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