Abnormal Stomatal Behavior and Hormonal Imbalance in flacca, a Wilty Mutant of Tomato: II. Auxin- and Abscisic Acid-like Activity

The wilty tomato mutant, flacca, and the control variety, Rheinlands Ruhm, were compared with regard to the endogenous activity and concentration of auxin- and abscisic acid-like substances during ontogeny. The mutant wilts fast under water deficit because of inability to close its stomata. Symptoms characteristic of excessive auxin are evident in the developing mutant. Among these symptoms are branch and leaf epinasty, excessive rooting along the stem, and increased apical dominance. By using a leucine-incorporation assay, spray of whole plants with 2,4-dichlorophenoxyacetic acid, and wheat coleoptile bioassay, indications were found of an excess of activity and concentration of auxin-like substances in shoots of young and mature mutant plants. The wheat coleoptile bioassay also revealed a much lower amount of substances with abscisic acid-like activity in the mutant compared with the normal plant. In contrast to the appearance during ontogeny of morphological symptoms characteristic of auxin excess in the mutant, the absolute amount of auxin-like substances and their activity in incorporation of leucine decreased with age. A parallel decrease of the concentration and activity of auxin-like compounds was also found in the normal plant. The concentration of abscisic acid-like substances increased with age in both genotypes. The disagreement between the increasing morphological symptoms and the decrease of auxin-like activity and concentration is discussed, together with the possibility of a causal relationship between auxin-and abscisic acid-like activity and stomatal behavior.     

Unrevealed structural requirements for auxin-like molecules by theoretical and experimental evidences

An computational-biostatistical approach, supported by ab initio optimizations of auxin-like molecules, was used to find biologically meaningful relationships between quantum chemical variables and fresh bioassay's data. It is proven that the auxin-like recognition requires different molecular assembling states. We suggest that the carboxyl group is not the determining factor in explaining the biological auxin-like conduct. The biological effects depends essentially on the chemical condition of the ring system. The aim to find active molecules (quantum objects) via statistical grouping-analysis of molecular quantum similarity measures was verified by bioactivity assays. Next, this approach led to the discovery of a non-carboxylated active auxin-like molecule (2,6-dibromo-phenol). This is the first publication on structure activity relationship of auxin-like molecules, which relies on highly standardized bioassays of different auxins screened in parallel as well as analysed by multi-dimensional scaling.     

Morphogenetic and cellular reorientations induced by Agrobacterium rhizogenes (strains 1855, 2659 and 8196) on carrot,pea and tobacco

The morphogenetic and cellular effects induced by three strains of Agrobacterium rhizogenes (1855, 2659 and 8196) inoculated on carrot root discs, pea epicotyls and tobacco stems have been investigated. Differential properties were observed according to bacterial strains, hosts and polarity. After inoculations with strain 1855, auxin-like symptoms have been detected in the reactive regions, at the cell level (cell expansion, nucleolar stimulation, DNA synthesis and endopolyploidization), during tissue reorganization (callogenesis, tracheogenesis, cambiogenesis) and new morphogenetic pathways (normal and transformed rhizogenesis, maintenance of apical dominance). Some of these symptoms were also observed with strain 2659 but not with strain 8196. Transformed roots were indirectly initiated from cambial-like layers redifferentiated inside a neoformed callus. While initial cortical cells of pea and, moreover, the subsequent callus, were partly polyploid, the resulting transformed roots were only diploid. The hypothesis of selection of diploid cells for transformation or root initiation is discussed as well as the observed auxin-like symptoms.     

Metabolism of Indole-3-Acetic Acid: II. Oxindole Pathway in Parthenocissus tricuspidata Crown-Gall Tissue Cultures

An indoleacetic acid oxidase preparation from an acetone powder of Parthenocissus tricuspidata crown-gall tissue has been examined. An intermediate in the reaction is 3-hydroxymethyloxindole and nonenzymic conversion of it to 3-methyleneoxindole was observed. Neither reaction mixtures nor 3-methyleneoxindole have any auxin-like activity in Avena or wheat coleoptile bioassays. In vivo studies show that although 53% decarboxylation of indoleacetic acid was observed in 48 hours, only a small amount of 3-methyloxindole could be recovered from the medium. The other decarboxylated products remain to be identified but are not 3-hydroxymethyloxindole or 3-methyleneoxindole.     

α-N-acetyl-indole-3-acetyl-ε-L-lysine: A metabolite of indole-3-acetic acid from Pseudomonas syringae pv. savastanoi

A product of indole-3-acetic acid (IAA) metabolism having an auxin-like activity has been isolated from liquid cultures of Pseudomonas syringae pv. savastanoi. By spectral data and chemical correlations the compound has been identified as α-N-acetyl-indole-3-acetyl-ε-l-lysine (Ac-IAA-Lys). The IAA-derivative was detected in culture filtrates of oleander strains but not in culture filtrates of olive strains. The physiological effects of Ac-IAA-Lys on hypocotyl elongation in wheat, leaf chlorosis in oleander and bean and the hypertrophic response of potato tuber discs were compared with those of IAA. The results indicated that Ac-IAA-Lys was approximately 60 % less active than IAA.     

Cytokinin- and auxin-like activity in Cyanophyta and microalgae

Seven strains of axenic unicellular Chlorophyta and three strains ofaxenic Cyanophyta were selected for their high growth rate and positivecytokinin-like activity in the Cucumber Cotyledon Expansion Bioassay. Ethanolicextracts of these ten strains were purified using cation exchange resin andpaper chromatography and then screened for cytokinin-like activity using theSoybean Callus Bioassay. No trend was detected in the Cyanophyta with onespecies having two peaks of cytokinin-like activity co-eluting with zeatinderivatives and iso-pentenyladenine derivatives and theother two species having only one peak of activity co-eluting with zeatinderivatives. Four Chlorophyta only had one peak of cytokinin-like activity,co-eluting with zeatin derivatives, while two species had two peaks of activityco-eluting with zeatin and iso-pentenyladeninederivatives.One Chlorophyta species had no detectable cytokinin-like activity. The same tenstrains were also tested for auxin-like activity. The water extract of sevenstrains demonstrated auxin-like activity in the Cucumber Cotyledon RootFormation Bioassay. No auxin-like activity was detected in any of the strainsusing the Mung Bean Bioassay after being extracted in methanol and purified byphase partitioning with phosphate buffer and ethyl acetate.     

Dominant lethal assay in female mice after oral dosing with dichlorvos or exposure to atmospheres containing dichlorovos.

The effect of a synthetic auxin-like substance (2,4-D) and a synthetic cell division factor (kinetin) on the induction of chromosome aberrations was studied on tissue cultures of Nicotiana glauca and the tumorous amphidiploid hybrid Nicotiana glauca × Nicotiana langsdorffii.The aberration frequencies in normal Nicotiana glauca tissue were proportional to the length of time of culture in the presence of 2,4-D. Moreover, both 2,4-D and kinetin increased chromosome breakage in the habiatouated Nicotiana glauca tissue but not in the amphidiploid hybrid tissue.The data are discussed in terms of genotype-hormone equilibria in long-term development of plant tissue culture.     

A comparative characterization of fungal melanin and the humin-like substances synthesized by <Emphasis Type="Italic">Cerrena maxima</Emphasis> 0275

Comparative studies of fungal melanin and two preparations of the high-molecular-weight humin-like substances formed during a solid-phase cultivation of the basidiomycete Cerrena maxima 0275 for 45 and 70 days were performed. The fungal melanin from Aspergillus niger and the humin-like substances synthesized by the basidiomycete C. maxima 0275 are similar in their physicochemical properties (elemental composition and behavior in acids and alkalis) and auxin-like activities. However, these biopolymers differ, essentially, at the structural level. According to IR spectroscopy data, the obtained humin-like substances display a higher similarity to natural humic acids and are more diverse in their functional groups compared with fungal melanins. Presumably, this is connected with the fact that laccase is involved in formation of humin-like substances; moreover, this enzyme is involved not only in the synthesis of these polymers, but also in their modification and degradation.     

Improvement of maize (Zea mays L.) anther culture responses by algae-derived natural substances

This is the first report on the beneficial effect of microalgal and cyanobacterial biomass on anther cultures of maize (Zea mays L.). Investigations were made on the cytokinin- and auxin-like activity and content of terrestrial and fresh-water living microalgal and cyanobacterial strains. The influence of media supplemented with biomass from four selected strains on the anther induction, the frequency of microspore-derived embryo-like structures, and regeneration capacity in anther cultures of maize was also studied. The addition of cyanobacterial and microalgal biomass to the induction and regeneration media in concentrations of 1 or 2 g/L improved the androgenic response, and was able to reduce the quantity of the synthetic auxin 2,4-dichlorophenoxy-acetic acid (2,4-D) required, or replace it completely.     

Auxin-like Growth Activity of Phenylacetonitrile

Phenylacetonitrile (PAN) and phenylacetic acid (PAA) showedauxin-like activity by stimulating elongation of wheat-coleoptilesections. Their auxin-like properties were also tested on theelongation of sections cut from hypocotyls of cress and sugar-beetseedlings, from petioles of sugar-beet leaves, and from epicotylsof pea seedlings. They were also tested for their ability toinduce adventitious roots on hypocotyls of cress and sugar-beetseedlings and on pea epicotyls. PAN was generally more activethan PAA on cress, and PAA more active than PAN on sugar-beetand peas. Steam distillates of cress seedlings contained a neutralsubstance with auxin-like activity and similar chromatographicproperties to PAN.     

Auxin-like Growth Activity of 3-Phenylpropionitrile from Water-Cress (Nasturtium officinale R. Br.)

Solutions of 3-phenylpropionitrile (PPN) and 3-phenylpropionicacid (PPA) exhibited auxin-like plant growth activity by stimulatingelongation of sections cut from wheat coleoptiles and garden-cresshypocotyls. Elongation of sugar-beet hypocotyl sections wasstimulated by PPA but not by PPN. Although PPA stimulated elongationof pea-epicotyl sections after 19 h incubation, most concentrationsof PPN did not show activity until 70 h. PPN generally stimulatedadventitious root formation on hypocotyls of garden-cress andsugar-beet, and on epicotyls of pea seedlings whereas dilutesolutions of PPA were inactive and concentrated solutions weretoxic. Steam distillates of water-cress plants contained a substancewith auxin-like growth activity and chromatographic propertiessimilar to PPN. Nasturtium officinale R. Br. water-cress, 3-phenylpropionitrile, 3-phenylpropionic acid, auxins     

Phloroglucinol in plant tissue culture

In plant tissue culture research, there is a constant need to search for novel substances that could result in better or more efficient growth in vitro. A relatively unknown compound, phloroglucinol (1,3,5-trihydroxybenzene), which is a degradation product of phloridzin, has growth-promoting properties. Phloroglucinol increases shoot formation and somatic embryogenesis in several horticultural and grain crops. When added to rooting media together with auxin, phloroglucinol further stimulates rooting, most likely because phloroglucinol and its homologues act as auxin synergists or auxin protectors. Of particular interest is the ability of phloroglucinol—a precursor in the lignin biosynthesis pathway—to effectively control hyperhydricity through the process of lignification, thus maximizing the multiplication rate of woody species and other species that are difficult to propagate. Phloroglucinol has also been used to improve the recovery of cryopreserved Dendrobium protocorms, increasing the potential of cryopreservation for application in ornamental biotechnology. Phloroglucinol demonstrates both cytokinin-like and auxin-like activity, much like thidiazuron, and thus has considerable potential for application in a wide range of plant tissue culture studies.     

Weakly cytokinin-active diphenylurea derivatives influence adventitious rooting in M26 Malus pumila microcuttings

The cytokinin activity of 8 previously unstudied diphenylurea (DPU)derivatives, differing in either type or position of the substituents of thephenyl rings, was investigated. Cytokinin activity was assessed using thebetacyanin (so-called amaranthin) accumulation test and the tomato regenerationtest. We also assayed their capacity to enhance adventitious root formation inmicrocuttings of apple rootstock M26 and in a tomato cotyledon rooting test,while their possible auxin-like activity was tested using the pea stemelongation test. All the compounds showed weak cytokinin activity, and thismight be responsible for the enhanced adventitious rooting which is induced bysome of the compounds. None of the DPU derivatives showed auxin-like activity.     

Characteristic features of the coumarin induced growth

Summary Effect of 2,4-dinitrophenol, iodoacetic acid, mannitol, ribonuclease, CaCl₂, phosphates, low oxygen tension, different temperatures, CCC and IAA on the coumarin-induced growth of sunflower hypocotyl sections have been investigated. Results indicate that coumarin-induced growth is auxin-like in type; nevertheless, the primary mode of coumarin action is probably quite different from that of IAA.With 4 Figures in the Text     

Auxin and gibberellin interactions in morphogenesis

Summary The effect of gibberellic acid on the biosynthesis of diphenolic compounds, growth and root initiation was investigated to determine a possible relationship between the development of morphological structures and a biochemical process. The results indicated that the effect of GA on growth was due to an increase in the amount of auxin-like substances.The mechanism by which a higher auxin level was brought about through treatment with gibberellin was shown not to involve inhibitors of IAA-oxidase but the synthesis of auxin. The level of anthocyanin and hydronaphtoquinone, which should be expected to inhibit IAA oxidation, does not affect the level of endogenous auxin-like substances. In contrast to a previous paper, it cannot be concluded that the rooting ability of balsam cuttings is determined by the level of indoleacetic acid.     

3-Phenyllactic acid is converted to phenylacetic acid and induces auxin-responsive root growth in Arabidopsis plants

Many microorganisms have been reported to produce compounds that promote plant growth and are thought to be involved in the establishment and maintenance of symbiotic relationships. 3-Phenyllactic acid (PLA) produced by lactic acid bacteria was previously shown to promote root growth in adzuki cuttings. However, the mode of action of PLA as a root-promoting substance had not been clarified. The present study therefore investigated the relationship between PLA and auxin. PLA was found to inhibit primary root elongation and to increase lateral root density in wild-type Arabidopsis, but not in an auxin signaling mutant. In addition, PLA induced IAA19 promoter fused β-glucuronidase gene expression, suggesting that PLA exhibits auxin-like activity. The inability of PLA to promote degradation of Auxin/Indole-3-Acetic Acid protein in a yeast heterologous reconstitution system indicated that PLA may not a ligand of auxin receptor. Using of a synthetic PLA labeled with stable isotope showed that exogenously applied PLA was converted to phenylacetic acid (PAA), an endogenous auxin, in both adzuki and Arabidopsis. Taken together, these results suggest that exogenous PLA promotes auxin signaling by conversion to PAA, thereby regulating root growth in plants.     

Synthesis,Crystal Structure and Biological Activity of 2-Hydroxyethylammonium Salt of p-Aminobenzoic Acid

p-Aminobenzoic acid (pABA) plays important roles in a wide variety of metabolic processes. Herein we report the synthesis, theoretical calculations, crystallographic investigation, and in vitro determination of the biological activity and phytotoxicity of the pABA salt, 2-hydroxyethylammonium p-aminobenzoate (HEA-pABA). The ability of neutral and anionic forms of pABA to interact with TIR1 pocket was investigated by calculation of molecular electrostatic potential maps on the accessible surface area, docking experiments, Molecular Dynamics and Quantum Mechanics/Molecular Mechanics calculations. The docking study of the folate precursor pABA, its anionic form and natural auxin (indole-3-acetic acid, IAA) with the auxin receptor TIR1 revealed a similar binding mode in the active site. The phytotoxic evaluation of HEA-pABA, pABA and 2-hydroxyethylamine (HEA) was performed on the model plant Arabidopsis thaliana ecotype Col 0 at five different concentrations. HEA-pABA and pABA acted as potential auxin-like regulators of root development in Arabidopsis thaliana (0.1 and 0.2 mM) and displayed an agravitropic root response at high concentration (2 mM). This study suggests that HEA-pABA and pABA might be considered as potential new regulators of plant growth.     

Hormonal Regulation of the Distribution of 14C-labelled Assimilates in the Flowering Shoot of Carnation

Evidence is presented that growth substances have a role indirecting the movement of assimilates towards the developingflowers of carnation. Removal of the terminal flower resultedin decreased upward movement of ¹⁴C-labelled assimilates froma source leaf on the flowering shoot. Applications of indole-3yl-aceticacid or gibberellic acid to the stump of the flower stem partiallyrestored the pattern of movement to that found in the intactshoot. Two gibberellin-like substances and one auxin-like substancewere detected in extracts of the floral tissues. Levels of growthsubstances appeared to be higher in the flower than elsewherein the flowering shoot.     

Mechanism of specific inhibition of phototropism by phenylacetic Acid in corn seedling

Using geotropism as a control for phototropism, compounds similar to phenylacetic acid that photoreact with flavins and/or have auxin-like activity were examined for their ability to specifically inhibit phototropism in corn seedlings using geotropism as a control. Results using indole-3-acetic acid, napthalene-1-acetic acid, naphthalene-2-acetic acid, phenylacetic acid, and β-phenylpyruvic acid suggest that such compounds will specifically inhibit phototropism primarily because of their photoreactivity with flavins and not their auxin activity. For example, strong auxins, indole-3-acetic acid and naphthalene-1-acetic acid, affected both tropic responses at all concentrations tested whereas weak auxins, phenylacetic acid and naphthalene-2-acetic acid, exhibited specific inhibition. In addition, the in vivo concentration of phenylacetic acid required to induce specificity was well below that required to stimulate coleoptile growth. Estimates of the percentage of photoreceptor pigment inactivated by phenylacetic acid (>10%) suggest that phenylacetic acid could be used to photoaffinity label the flavoprotein involved in corn seedling phototropism.