Relazioni ormonali nello sviluppo dell'ovario delle orchidee II. Azione delle auxine in segmenti isolati dell'asse fiorale

Abstract

HORMONAL RELATIONS IN THE DEVELOPMENT OF ORCHID OVARY. II. — The effects of auxins on isolated parts of the flower axis. — The changes in fresh weight of segments of column, ovary and peduncle of Cymb[icaron]dium sp., treated in vitro with auxins (IAA, EtIA and NAA) indicate that these parts of the flower react promptly and independently to the three auxins tested. Expansion is very small in the ovary, and is more and more large moving towards the apex of the flower (fig. 1). The apical part of the column reacts somewhat more rapidly to auxin and tolerates higher concentration than the proximal part.

Naphtalenacetic acid proved to be the most active of the three auxins tested. This finding is in agreement with the higher efficiency of this substances as an inducer of parthe-nocarpy and its ability to induce large necroses of the column, when applied to the flower on the intact plant.

The plots of auxin activity against concentration of the auxins suggest thai the lower activity of IAA might be due to its destruction by indoleacetic acid oxidase.     

Effetti del trattamento In Vivo con auxlna di internodi di pisello sulla fosforilazione ossidativa dei mitocondri In Vitro

Abstract

EFFECT OF TREATMENT WITH IAA OF PEA INTERNODE SECTIONS ON THE OXIDATIVE PHOSPHORYLATION OF THEIR MITOCHONDRIA. — It was previously shown that auxin treatment raises the level of ATP in pea stem sections, under the conditions where the hormone stimulates growth (MARRé AND FORTI). It is also known that under the same conditions auxin stimulates oxygen uptake (MARRé, FORTI e ARRIGONI; MARRÉ and FORTI), and that the auxin induced respiration is most probably mediated by cytochrome oxidase (MARRÉ, FORTI and GAUR).

However, auxin has no effect when added to isolated mitochondria. In this paper, the effect of auxin treatment of the tissue on the activity of mitochondria isolated after the hormone treatment has been studied. It has been found that oxidative phosphorylation of mitochondria from the auxin treated pea stem sections is 13% higher than that of controi sections. The auxin effect is significant at 96% probabilities. There is no effect of the hormone on the P/O ratio.     

Rinvenimento del polline di Keteleeria in alcuni depositi quaternari dell'Italia centro-settentrionale

Abstract

THE DISCOVERING OF THE KETELEERIA'S POLLEN-GRAIN IN SOME QUATERNARY DEPOSITS OF THE CENTRAL AND NORTHERN ITALY. — The Author observes the specy of Keteleeria with the pollen analysis executed on deposits of turfys, clays and brown coals of some Umbro-Marchigiano and of the Po Plain localyties.

The Keteleeria to-day is living only in China and Japan; in Italy it was living also during the early and middle Pleistocene.     

The role of auxins in differentiation of rice tissues culturein Vitro

Effects of various auxins on callus induction (dedifferentiation) and organ redifferentiation from the callus were studied by using various tissues of rice,Oryza sativa L. cv. Kyoto Asahi. 2,4-D, NAA and IAA were used as auxins for the test of their ability to induce callus. All of these were active. This callus induction by auxin was successful in all tissues used; seed, root, shoot nodule, anther and ovary. In all of the calluses induced by various auxins such as 2,4-D, NAA and IAA and derived from various tissues such as seed, root, shoot nodule, anther and ovary, organ redifferentiation, i.e., formation of shoots and roots was achieved by removing the auxins from the medium used for the callus calture. Cytokinins were not necessary for the organ redifferentiation in these calluses. These results suggest that auxin is the only exogenous factor that determines dedifferentiation and redifferentiation in rice plant tissues culturedin vitro.     

Istogenesi e caratteri anatomici dei tessuti ricavati da frammenti di germoglio di Solanum Tuberosum L. coltivati In Vitro

Abstract

HISTOGENESIS AND ANATOMICAL CHARACTERISTICS OF PORTIONS OF SOLANUM TUBEROSUM SPROUTS CULTIVATED IN VITRO. — Disks cut from sprouts of Solanum tuberosum L. have been cultured on White's basic or modified medium (White, 1943) with 20 mg/l KH2₂PO₄ 50 mg/l adenine, and 12 or 24 mg/l NAA.

Sections were prepared according to Morel's method (1948). Only on the disks cultured on White's modified medium with 12 mg/1/NAA growth glomerules and a normal meristematic layer were present, while there were no signs of cellular hypertrophy and organogenetic phenomena.

12 mg/l appears therefore to be the optimal dose of NAA for cultures of excised portions of potato sprouts. Adenine is however always necessary because it triplicates the growth activity of the tissues and because through its antagonism with regard to auxin it favours the genesis of the callus.     

Osservazioni carioembriologiche in Ornithogalum di Sardegna: Nota I. - Sporogenesi e sviluppo dei gametofiti in Ornithogalum caudatum Ait

Abstract

CARYOEMBRYOLOGICAL STUDIES in ORNITHOGALUM OF SARDINIA.

I. - SPOROGENESIS AND DEVELOPMENT OF THE GAMETOPHYTES IN ÒRNITHOGALUM CAUDATUM AIT. — After a concise synthesis of the caryoembryological knowledges regarding the taxa of Ornithogalum present in Sardinia, the embryology and morphology of Ornithogalum caudatum Ait., have been investigated.

In a three-locular ovary the ovules are anatropous, epitropous with nucellus crassinucellate and only one archesporial cell evolves a mother cell.

The development of the female gametophyte tallies with the Polygonum (Normal) Type.

The homoetypical division presents an asynchronous proceeding in the cellules of the dyad with a considerable delay in the two-nucleate arrangement, especially in the chalazal cell.

Antipodal cells present a hypertrophic growing, but they present no tendency to polyantipody.

The endosperm is of the Helobiae Type.

The divisions of the pollen mother-cells are of the successive Type.

Longest axis of 1-sulcate pollen grains about 76 micron.

A true tapetal periplasmodium isn't formed in the anthers.

The chromosome number is 2n = 54.

Raphides accour in the cataphylls, in the parenchymatous tissues of the leaf and in several parts of the floral region.     

Interactions of auxin and gibberellin in the control of basal growth of <Emphasis Type="Italic">Arabidopsis</Emphasis> rosette leaves

Arabidopsis is a species that naturally displays the rosette form. Therefore, elucidation of the factors, which control basal leaf development, is of particular interest. Most evidence points that auxins and gibberellins are important in the control of rosette leaf development. In this paper, we report on a regimen that disrupts the normal rosette growth in Arabidopsis and induces internodal growth, which we have termed unbasal. The growth conditions are: (1) seed germination in the presence of 2,3,5-triiodobenzoic acid (TIBA); (2) transfer of the seedlings to a medium containing exogenous auxin (NAA) and GA₃; (3) transfer of the seedlings to a GA₃-only medium for all subsequent growth. Under these conditions, auxin and GA interact to induce internode elongation. Polar auxin transport appears to have a temporal effect on this synergistic interaction. In this regimen, GA increases auxin activity in the basal portions of the stem. Cross sectional morphology of the elongated internodes between two rosette leaves in an un-basal plant was similar to that seen for the pin1 Arabidopsis mutation.     

Hormonal Effects on Growth and Morphology of Normal and Hairy Roots of Hyoscyamus muticus

Treatment of normal and Agrobacterium rhizogenes-transformed root cultures of Hyoscyamus muticus with three different auxins, indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and naphthaleneacetic acid (NAA), revealed that the response varied considerably among auxins, between transformed and normal roots, and depending on the parameter. In normal roots all three auxins provoked abundant branching, with IBA and NAA being the most effective at 2.5 and 0.5 μm, respectively, whereas IAA was most effective at low concentrations (0.05 and 0.1 μm). In transformed roots exogenously supplied auxins were generally inhibitory or, at best, without effect on growth and branching. Only 0.01 μm IAA significantly enhanced lateral root number, whereas at the higher concentrations IBA, although inhibitory, was the least effective auxin. In both root types IBA had little effect on primary root growth, but normal roots were more sensitive to IAA and NAA. These results suggest a different sensitivity to auxins of normal and transformed roots since there was no significant difference in endogenous free and conjugated IAA content nor in IAA uptake capacity. Ethylene production and biosynthesis were approximately threefold higher in hairy roots, but production could be stimulated up to tenfold that of control levels in normal roots by supplying NAA or 1-aminocyclopropane-1-carboxylic acid (ACC). Treatment with 2.5 μm NAA, but not IAA or IBA, also enhanced ethylene biosynthesis in normal roots but not in transformed ones. ACC and malonyl-1-aminocyclopropane-1-carboxylic acid accumulated to detectable levels only after treatment with an auxin (NAA). Received March 3, 1997; accepted May 28, 1997     

Isolierung von Auxinen aus dem Chromatin regenerierender Erbsenkeimlinge

Summary Etiolated pea epicotyls were treated with IAA, NAA or 2,4-D for 48 hours. From the chromatin (purified by gel-chromatography) of the treated seedlings growth substances could be isolated which significantly promote growth of Avena coleoptiles. The isolated growth substances showed the same RF as the applied auxins. When the seedlings were treated for 48 hours either with auxin or with water only the extract from the auxin treated plants promoted Avena growth significantly. The ether extract from the auxin treated seedlings still had growth promoting effect when the seedlings were treated 24 hours with auxin and then 24 hours with water to remove most of the free auxin in the cells and in the cell walls.     

The application of growth regulators for the physiological emasculation of rye(secale cereale l.) flowers

The present paper deals with the possibility of using growth regulators for the physiological emasculation of rye flowers and utilizes our knowledge of the higher sensibility of stamens to the auxin level. By means of bioassays it has been found that a relatively low content of auxins and a definite level of gibberellins and inhibitors are characteristic for normal rye spikes at the time of stamen differentiation. The higher level of auxins and expressive inhibitions occur in the later course of pistil differentiation. Two sprayings of plants with mixed solutions of MH and NAA and two further sprayings with a solution of NAA only before the stamen differentiation change slightly the natural character of endogenous regulators in spikes and cause the destruction of mother pollen cells in anthers and the sterility of anthers in the majority of rye florets. The anthers in the basis of spikes sometimes contain pollen grains after treatment, but their fertility is substantially lowered because the spray evidently disturbed the accumulation of reserve substances in pollen grains. The development of pistil was not affected by the exogenous application of growth regulators. Normal kernels arose in the majority of flowers after supplementary pollination. Some accidental effects of exogenous application of growth regulators, as for example reduced stem growth and disorder of lodicule function were lowered to a great extent by adding GA_S to the last two sprayings of NAA.     

Genotypic Responses to Auxins in Tissue Cultures of Phaseolus

Responses of seven Phaseolus genotypes to the four auxins picloram, 2,4-D, NAA and 1AA, in tissue cultures, were examined. Callus growth was promoted by picloram and the range of effective concentrations for most genotypes was broad. The auxin 2,4-D also enhanced callus growth, but the range of optimal concentrations was markedly narrower than that of picloram. NAA when supplied at relatively high concentrations gave good growth. IAA was ineffective in supporting callus growth. The differences in 2,4-D concentrations required for optimal growth and the differential responses to low concentrations (0.04–1.25 μM) of picloram between several genotypes tested were large. These genotypic variations in auxin responses were repeatable and may thus reflect genetic differences.     

Metabolic changes during rooting in stem cuttings of five mangrove species

Vegetative propagation through rooting in stem cuttings in five tree mangroves namely Bruguiera parviflora, Cynometra iripa, Excoecaria agallocha, Heritiera fomes, and Thespesia populnea using IAA, IBA and NAA was reported. Spectacular increase in the root number was noted in the cuttings of H. fomes and C. iripa treated together with IBA (5000 ppm) and NAA (2500 ppm). The highest number of roots was obtained with IBA (2500 ppm) and NAA (500 ppm) in E. agallocha. B. parviflora and T. populnea responded better to IAA and IBA treatment. The species specific variation in the rooting response to exogenous application of auxins was reflected in the metabolic changes during initiation and development of roots in cuttings. Biochemical analysis showed increase of reducing sugar in the above-girdled tissues at initiation as well as subsequent development of roots which was further enhanced by the use of auxins. Decreases in the total sugar, total carbohydrate and polyphenols and increase in total nitrogen were recorded in the girdled tissues and the high C/N ratio at the initial stage helped in initiation of roots in all the species. Interaction of IBA and NAA promoted starch hydrolysis better than IAA and IBA during root development and subsequently reduced the C/N ratio and increased the protein-nitrogen activity during root development which suggest the auxin influenced mobilization of nitrogen to the rooting zone.Abbreviations IAA Indole-3-acetic acid - IBA Indole-butyric acid - NAA A-naphthalene acetic acid     

The effects of auxin on lateral root initiation and root gravitropism in a lateral rootless mutant Lrt1 of rice (Oryza sativa L.)

Auxins control growth and development in plants, including lateral rootinitiation and root gravity response. However, how endogenous auxin regulatesthese processes is poorly understood. In this study, the effects of auxins onlateral root initiation and root gravity response in rice were investigatedusing a lateral rootless mutant Lrt1, which fails to formlateral roots and shows a reduced root gravity response. Exogenous applicationof IBA to the Lrt1 mutant restored both lateral rootinitiation and root gravitropism. However, application of IAA, a major form ofnatural auxin, restored only root gravitropic response but not lateral rootinitiation. These results suggest that IBA is more effective than IAA in lateralroot formation and that IBA also plays an important role in root gravitropicresponse in rice. The application of NAA restored lateral root initiation, butdid not completely restore root gravitropism. Root elongation assays ofLrt1 displayed resistance to 2,4-D, NAA, IBA, and IAA.This result suggests that the reduced sensitivity to exogenous auxins may be due tothe altered auxin activity in the root, thereby affecting root morphology inLrt1.     

Effects of Natural and Synthetic Auxins on the Gravitropic Growth Habit of Roots in Two Auxin-Resistant Mutants of Arabidopsis, axr1 and axr4: Evidence for Defects in the Auxin Influx Mechanism of axr4

Arabidopsis thaliana, axr4 , was restored by the addition of 30–300 nM 1-naphthaleneacetic acid (NAA) to the growth medium. Neither indole-3-acetic acid (IAA) nor 2,4-dichlorophenoxyacetic acid (2,4-D) showed such an effect. Growth of axr4 roots was resistant to IAA and 2,4-D, but not at all to NAA. The differential effects of the three auxins suggest that the defects of axr4 result from a lower auxin influx into its cells. The partially agravitropic growth habit of axr1 roots, which was less severe than that of axr4 roots, was only slightly affected by the three auxins in the growth medium at concentrations up to 300 nM; growth of axr1 roots was resistant to all three of the auxins. These results suggest that the lesion of axr1 mutants is different from that of axr4. Received 9 June 1999/ Accepted in revised form 16 August 1999     

Chemical induction of adventitious root formation in Taxus baccata cuttings

The effect of some auxins (IBA and NAA), phenolic compounds (phloroglucinol, gentisic acid and coumarin), a combination of auxins and phenolics, and a systemic fungicide (Bavistin) have been examined for stimulatory effects on adventitious root formation in stem cuttings (current season's growth) of Taxus baccata L. In general lower concentration (0.25 mM) of both IBA and NAA was more effective in inducing rooting of cuttings taken from both male and female trees. The combined treatment of IBA+NAA (0.25 mM each) showed some success in cuttings from male trees only (55%, compared to 15% rooting in cuttings from female trees). Generally, the callus formation was quite high (70%) in all auxin treatments (alone or in combination). Among the phenolics, 40% rooting success was achieved with phloroglucinol only, while coumarin and gentisic acid were ineffective. The combined treatment of auxins and phenolics also failed to promote rooting. On the other hand, Bavistin was extremely effective for callusing (90%) as well as rooting (80%). The effectiveness of various compounds tested for rooting of young stem cuttings declined in the order: 0.25 mM IBA>0.05% Bavistin>0.25 mM NAA>1.25 mM IBA>15 mM phloroglucinol>IBA+NAA (0.25 mM each). In addition to the auxins, IBA and NAA that are widely used for commercial propagation, the auxin-like properties of the fungicide Bavistin could be exploited for adventitious rooting in T. baccata, and in other plant species.     

Modified hormonal balance in rooting-recalcitrant rac mutant tobacco shoots

ABSTRACT

The rooting-recalcitrant rac tobacco mutant has been multiplied in vitro via outgrowth of axillary buds in parallel to the D8 wild-type. The mutant shoots grew at a lower rate and did not root whatever the treatments, whereas the wild-type shoots rooted spontaneously during the culture cycle without auxin treatment. The mutant and wild-type shoots showed similar peroxidase variations along the culture cycle (21 days) but with higher levels of activity for the rac mutant: minimum peroxidase activity occurred at day 14 in whole shoots of both tobacco genotypes, but already at day 7 in the basal parts of the stems (where roots appear) of the wild-type tobacco, while it was delayed in the mutant. Free and conjugated auxin and polyamine levels were also determined in whole shoots and basal parts of the stems. The rac mutant was characterised by higher auxin and polyamine contents. A peak of auxins and polyamines appeared at day 14 in the whole shoots whatever the tobacco genotype. This peak was delayed in the basal parts of the rac stems compared to the wild-type ones. The mutant shoots contained higher levels of benzyladenine and isopentenyladenosine at the end of the culture cycle, whereas zeatin riboside was more abundant in wild-type shoots. In response to increasing concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA), only the wild-type shoots responded by an increase in growth rate followed by inhibition at high concentrations. The rac shoot responses were very low or nonexistent. Peroxidase activity was also measured in the basal parts of tobacco stems grown in the presence of IBA. Results suggest growth inhibition related to auxin accumulation, possibly combined with elevated putrescine content. Second, rooting induction seems to take place in both tobacco genotypes; however, the process of root formation is blocked in the mutant. The lack of initiation and expression phases of rooting in relation to auxin content in the mutant is discussed.     

Influence of galactoglucomannan oligosaccharides on root culture of <Emphasis Type="Italic">Karwinskia humboldtiana</Emphasis>

Galactoglucomannan oligosaccharides (GGMOs) activity in K. humboldtiana root culture has been determined. GGMOs inhibited adventitious root growth and lateral root induction in contrast to IAA, IBA, and NAA stimulating effect in these processes. Similarly, the combination of GGMOs with natural auxins (IAA, IBA) evoked an inhibition of adventitious root growth and lateral root induction that depended on the oligosaccharides concentration and the type of auxin. The growth stimulating effect of the synthetic auxin, NAA, in adventitious roots was negatively affected by GGMOs, but they were without influence on lateral root induction. The presence of oligosaccharides triggered lateral root position on adventitious roots and the anatomy of adventitious roots (diameter, proportion of primary cortex to the central cylinder, number and size of primary cortical cells, intercellular spaces, and the number of starch grains in cells of primary cortex) in dependence on their coactions with auxin.     

Veränderungen des Nucleoproteids von Erbsenepikotylen durch synthetische Auxine bei der Induktion der Wurzelneubildung

Summary Root initiation in etiolated pea epicotyls induced by NAA and 2,4-D is inhibited by application of 5-BU in the same way as root initiation induced with IAA. The time of 5-BU action is regulated by auxin concentration. Binding of auxins to carefully extracted nucleoproteins from pea epicotyls at pH 8 resulted in a decrease of the melting point (T _m) of the nucleoproteins. From the results it is concluded that auxins act by direct binding to the nucleoprotein (probably by ionic bonds) and that this binding induces a decrease in binding capacity of histone to DNA, which represents the first step in the initiation of RNA synthesis.     

Effect of auxins and plant oligosaccharides on root formation and elongation growth of mung bean hypocotyls

The interaction of auxins – IAA, IBA or NAA – with galactoglucomannan oligosaccharides (GGMOs) on adventitious root formation and elongation growth of mung bean hypocotyl cuttings was studied. GGMOs induced adventitious roots in the absence of auxins; however, their effect was lower compared with IBA or NAA. On the other hand, in the presence of auxins, GGMOs inhibited adventitious root induction. Their effect depended on the concentration of oligosaccharides and the type of auxin used. The highest inhibition effect of GGMOs at a concentration of 10⁻⁸ M in the presence of IBA and NAA was observed. In the presence of IAA their inhibition was non-significant in regard to the concentration. The interaction of auxins with GGMOs resulted in the formation of adventitious roots on a shorter part of hypocotyls compared with the effect of auxins alone. However, roots were induced more extensively along the hypocotyls treated with GGMOs compared with the control. GGMOs inhibited the length of induced adventitious roots in the presence of IAA, while in combination with IBA or NAA they were ineffective. The elongation of hypocotyls induced by IAA or IBA was inhibited by GGMOs, too. However, in the presence of NAA or by endogenous growth they were without any significant effect on elongation growth. These findings suggest that GGMOs in certain concentrations might inhibit rooting and the elongation process dependant on auxin used.     

Effects of plant growth regulators on acetylene-reducing associations between Azospirillum brasilense and wheat

Treatment of wheat seedlings with the synthetic auxin, 2,4-dichlorophenoxyacetic acid (2,4-d), induced nodule-like structures or tumours (termed para-nodules) where lateral roots would normally emerge. The formation of these structures promoted increased rates of acetylene reduction at reduced oxygen pressure (0.02–0.04 atm) in seedling inoculated with Azospirillum brasilense, compared to seedlings inoculated without auxin treatment. Fluorescent microscopy, laser scanning confocal microscopy and direct bacterial counts all showed that the 2,4-d treatment stimulated internal colonization of the root system with azospirilla, particularly in the basal region of the nodular structures. Both colonization with azospirilla and acetylene-reducing activity were further stimulated by simultaneous treatment with another synthetic auxin, naphthaleneacetic acid (NAA) and, less reliably, with indoleacetic acid (IAA) and indolebutyric acid (IBA). These auxins produced shortening of many initiated lateral roots, although 20 times the concentration of NAA was required to achieve rounded structures similar to those obtained with 2,4-d. Treatment with NAA, IAA or IBA alone also stimulated colonization with azospirilla and acetylene reduction rates at 0.02 atm oxygen, but less effectively than by treatment with 2,4-d. Such exogenous treatments of wheat seedlings with synthetic growth regulators provide an effective laboratory model for studies on the development of a N₂-fixing system in cereals.