Inhibitory effect of a rhizobitoxine analog on bud growth after release from dormancy

Application of the ethoxy analog of rhizobitoxine (l-2-amino-4-[2'-aminoethoxy]-trans-3-butenoic acid), an inhibitor of ethylene biosynthesis, inhibited growth of apple, crabapple, and apricot buds released from dormancy by chilling or by treatment with benzyladenine. When tea crabapple (Malus hupehensis [Pamp.] Rehd.) buds were sprayed once with 8.8 x 10(-3)m benzyladenine, ethylene production by the buds increased significantly 24 to 48 hours after benzyladenine treatment. Application of the rhizobitoxine analog to the buds at the time of benzyladenine treatment reduced ethylene evolution to the level of the controls for up to 2 weeks after treatment. Increase in bud weight was inhibited also but to a lesser extent. These data suggest that growth of buds is accompanied by ethylene production and that the inhibition of ethylene biosynthesis also inhibits bud growth. Since additional metabolic effects result from the action of the rhizobitoxine analog, no firm conclusions on its role can be drawn at this time.     

Hormonal Regulation of Cell Elongation in the Hypocotyl of Rootless Soybean: An Evaluation of the Role of DNA Synthesis

A method was developed where soybean seedlings were grown without roots to study the influence of hormones of root origin on shoot growth. Excision of the root resulted in inhibition of apical section growth and DNA synthesis and inhibited elongating section growth. A synthetic cytokinin restored DNA synthesis in the apical section, but did not influence growth in either the apical or elongating sections. Low concentrations of gibberellin with the cytokinin restored growth in the apical section. Gibberellin alone was sufficient to restore growth in the elongating section.An inhibitor of DNA synthesis, 5-fluorodeoxyuridine, inhibited the increase in apical section DNA without inhibiting control or gibberellin-induced growth in the elongating section. Experiments with (14)C-thymidine resulted in no DNA labeling differences in the elongating section under conditions where gibberellin-induced elongation varied from 50% to 73% above controls. It was concluded that gibberellin-induced elongation in soybean hypocotyl occurred in the absence of DNA synthesis. Gibberellin does stimulate DNA synthesis in the apical tissue apart from its effect on cell elongation.Excised soybean hypocotyl elongated maximally at 10(-6)m auxin. At higher auxin concentrations, fresh weight and ethylene production increased, but elongation was reduced. Addition of GA to the higher auxin concentrations resulted in a 50% inhibition in auxin-induced ethylene production and resumption in maximal elongation. Added ethylene inhibited elongation 30% at 2 mul/l. Addition of up to 100 mul/l ethylene did not inhibit elongation with GA present in the incubation medium. Thus GA may counteract ehtylene inhibition of cell elongation in addition to inhibiting ethylene production in auxin-treated tissues.     

Endogenous Auxin is Required but Supraoptimal for Rapid Growth of Rice (Oryza sativa L.) Seminal Roots, and Auxin Inhibition of Rice Seminal Root Growth is Not Caused by Ethylene

The dual effects of auxin and ethylene on rice seminal root growth were investigated in this study. Low concentrations of exogenous indole-3-acetic acid (IAA) had no effect on rice seminal root growth, whereas higher concentrations (≥0.003 μM) were inhibitory. In contrast, low concentrations of the auxin action inhibitor p-chlorophenoxyisobutyric acid (PCIB), ranging from 0.5 to 50 μM, promoted rice seminal root growth, whereas high concentrations of PCIB (≥500 μM) and the polar auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA) inhibited rice seminal root growth. These results suggest that endogenous auxin is required but supraoptimal for rapid growth of rice seminal roots. In addition, although rice seminal root growth was inhibited by the exogenous ethylene-releasing compound ethephon or the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) as well as exogenous IAA, the 50% inhibition of growth (I₅₀) caused by ethephon or ACC was weakened by certain concentrations of the ethylene action inhibitor Ag⁺ (0.016-0.4 μM). However, the I₅₀ caused by exogenous IAA was strengthened by Ag⁺ or the ethylene biosynthetic inhibitor aminoethoxyvinylglycine (AVG) and weakened by certain concentrations of PCIB (0.5-50 μM). Together, the inhibitory mechanisms of auxin and ethylene on rice seminal root growth should be different, and auxin inhibition of rice seminal root growth should not be caused by ethylene. Furthermore, our results indicated that a certain threshold level of ethylene was required to maintain rice seminal root growth, and that ethylene within the threshold may antagonize auxin inhibition of rice seminal root growth.     

Rhizobitoxine production by Bradyrhizobium elkanii enhances nodulation and competitiveness on Macroptilium atropurpureum

Application of 1-aminoocyclopropane-1-carboxylic acid, an ethylene precursor, decreased nodulation of Macroptilium atropurpureum by Bradyrhizobium elkanii. B. elkanii produces rhizobitoxine, an ethylene synthesis inhibitor. Elimination of rhizobitoxine production in B. elkanii increased ethylene evolution and decreased nodulation and competitiveness on M. atropurpureum. These results suggest that rhizobitoxine enhances nodulation and competitiveness of B. elkanii on M. atropurpureum.     

An Ethylene-Mediated Increase in Sensitivity to Auxin Induces Adventitious Root Formation in Flooded Rumex palustris Sm

The hormonal regulation of adventitious root formation induced by flooding of the root system was investigated in the wetland species Rumex palustris Sm. Adventitious root development at the base of the shoot is an important adaptation to flooded conditions and takes place soon after the onset of flooding. Decreases in either endogenous auxin or ethylene concentrations induced by application of inhibitors of either auxin transport or ethylene biosynthesis reduced the number of adventitious roots formed by flooded plants, suggesting an involvement of these hormones in the rooting process. The rooting response during flooding was preceded by increased endogenous ethylene concentrations in the root system. The endogenous auxin concentration did not change during flooding-induced rooting, but a continuous basipetal transport of auxin from the shoot to the rooting zone appeared to be essential in maintaining stable auxin concentrations. These results suggest that the higher ethylene concentration in soil-flooded plants increases the sensitivity of the root-forming tissues to endogenous indoleacetic acid, thus initiating the formation of adventitious roots.     

Inhibitory action of auxin on root elongation not mediated by ethylene

The inhibitory effects of indole-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) on elongation growth of pea (Pisum sativum L.) seedling roots were investigated in relation to the effects of these compounds on ethylene production by the root tips. When added to the growth solution both compounds caused a progressively increasing inhibition of growth within the concentration range of 0.01 to 1 micromolar. However, only ACC increased ethylene production in root tips excised from the treated seedlings after 24 hours. High auxin concentrations caused a transitory increase of ethylene production during a few hours in the beginning of the treatment period, but even in 1 micromolar IAA this increase was too low to have any appreciable effect on growth. ACC, but not IAA, caused growth curvatures, typical of ethylene treatment, in the root tips. IAA caused conspicuous swelling of the root tips while ACC did not. Cobalt and silver ions reversed the growth inhibitory effects induced by ACC but did not counteract the inhibition of elongation or swelling caused by IAA. The growth effects caused by the ACC treatments were obviously due to ethylene production. We found no evidence to indicate that the growth inhibition or swelling caused by IAA is mediated by ethylene. It is concluded that the inhibitory action of IAA on root growth is caused by this auxin per se.     

Characterization of 1-aminocyclopropane-1-carboxylate (ACC) deaminase containing <Emphasis Type="Italic">Methylobacterium oryzae</Emphasis> and interactions with auxins and ACC regulation of ethylene in canola (<Emphasis Type="Italic">Brassica campestris</Emphasis>)

The possible interaction of the plant hormones auxin and ethylene and the role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase containing bacteria on ethylene production in canola (Brassica campestris) in the presence of inhibitory concentrations of growth regulators were investigated. The effects of auxin (indole-3-acetic acid and 2,4-dichlorophenoxy acetic acid), auxin transport inhibitor 2-(p-chlorophenoxy)-2-methylpropionic acid, ethylene precursor 1-aminocyclopropane-1-carboxylate and ethylene synthesis inhibitor l-α-(2-aminoethoxyvinyl)glycine hydrochloride on root elongation were concentration dependent. Exogenous addition of growth regulators influences the enzyme activities of ethylene production and we have presented here evidences that support the hypothesis that inhibitory effects of auxin on root elongation are independent of ethylene. Additionally, we have proved that inoculation of ACC deaminase containing Methylobacterium oryzae sequester ACC exuded from roots and hydrolyze them lowering the concentration of ACC in root exudates. However, the inhibitory actions of exogenous additions of auxins could not be ameliorated by bacterial inoculation that reduces ethylene concentration in canola seedlings.     

Factors increasing ethylene production enhance the sensitivity of root growth to auxins

Light inhibits root elongation, increases ethylene production and enhances the inhibitory action of auxins on root elongation of pea ( Pisum sativum L. cv. Weibulls Marma) seedlings. To investigate the role of ethylene in the interaction between light and auxin, the level of ethylene production in darkness was increased to the level produced in light by supplying 1-aminocyclopropane-1-carboxylic acid (ACC) or benzylaminopurine (BAP). Ethylene production was measured in excised root tips after treatment of intact seedlings for 24 h, while root growth was measured after 48 h. Auxin, at a concentration causing a partial inhibition of root elongation, did not increase ethylene production significantly. A 4-fold increase in ethylene production, caused either by light, 0.1 μ M ACC or 0.1 μ M BAP, inhibited root elongation by 40–50%. The auxins 2,4-dichlorophenoxyacetic acid and indolebutyric acid applied at 0.1 μ M inhibited root elongation by 15–25% in darkness but by 50–60% in light. Supply of ACC or BAP in darkness enhanced the inhibitory effects of auxins to about the same extent as in light. The inhibition caused by the auxins as well as by the BAP was associated with swelling of the root tips. ACC and BAP treatment synergistically increased the swelling caused by auxins. We conclude that auxin and ethylene, when applied or produced in partially inhibitory concentrations, act synergistically to inhibit root elongation and increase root diameter. The effect of light on the response of the roots to auxins is mediated by a light-induced increase in ethylene production.     

Ethylene upregulates auxin biosynthesis in Arabidopsis seedlings to enhance inhibition of root cell elongation

Ethylene represents an important regulatory signal for root development. Genetic studies in Arabidopsis thaliana have demonstrated that ethylene inhibition of root growth involves another hormone signal, auxin. This study investigated why auxin was required by ethylene to regulate root growth. We initially observed that ethylene positively controls auxin biosynthesis in the root apex. We subsequently demonstrated that ethylene-regulated root growth is dependent on (1) the transport of auxin from the root apex via the lateral root cap and (2) auxin responses occurring in multiple elongation zone tissues. Detailed growth studies revealed that the ability of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid to inhibit root cell elongation was significantly enhanced in the presence of auxin. We conclude that by upregulating auxin biosynthesis, ethylene facilitates its ability to inhibit root cell expansion.     

Studies on plant growth-regulating substances: XXXV. Ethylene production by coleoptiles treated with auxin-type chemicals

Wheat coleoptile sections were treated with a range of auxins and with compounds of related chemical structure which do not exhibit auxin properties. Methods used for measuring the rates of elongation and ethylene evolution of these sections are described. Ethylene was evolved some time after elongation in all cases and increased ethylene production occurred only with compounds showing auxin activity. The results indicate that ethylene evolution was related exponentially to growth. Simultaneous applications of mannitol and 2, 4-dichloro-phenoxyacetic acid (2, 4-D) to wheat sections markedly reduced ethylene evolution compared with the 2, 4-D controls, even though the level of 2, 4-D in the tissue apparently remained unchanged. Ethylene significantly inhibited the elongation of wheat coleoptile sections, and it is suggested that ethylene is a natural plant growth inhibitor which becomes mobilised to limit excessive growth.     

Auxin is a positive regulator for ethylene-mediated response in the growth of Arabidopsis roots

The requirement of auxin for the ethylene-mediated growth response in the root of Arabidopsis thaliana seedlings was investigated using two ethylene-resistant mutants, aux1-7 and eir1-1, whose roots have been shown to have a defect in the auxin influx and efflux carriers, respectively. A 50% inhibition of growth (I(50)) was achieved with 0.84 microl liter(-1) ethylene in wild-type roots, but 71.3 microl liter( -1) ethylene was required to induce I(50) in eir1-1 roots. In aux1-7 roots, I(50) was not obtained even at 1,000 microl liter(-1) ethylene. By contrast, in the presence of 10 nM 1-naphthaleneacetic acid (NAA), the concentrations of ethylene required to induce I(50) in eir1-1 and aux1-7 roots were greatly reduced nearly to the level required in wild-type roots. Since the action of NAA to restore the ethylene response in aux1-7 roots was not replaced by IAA, an increase in the intracellular level of auxin is likely to be the cause for the restoration of ethylene response. NAA at 10 nM did not inhibit root growth when applied solely, but it was the optimum concentration to recover the ethylene response in the mutant roots. These results suggest that auxin is a positive regulator for ethylene-induced inhibition in root elongation.     

Rhizobitoxine Production by Bradyrhizobium elkanii Enhances Nodulation and Competitiveness on Macroptilium atropurpureum

Application of 1-aminoocyclopropane-1-carboxylic acid, an ethylene precursor, decreased nodulation of Macroptilium atropurpureum by Bradyrhizobium elkanii. B. elkanii produces rhizobitoxine, an ethylene synthesis inhibitor. Elimination of rhizobitoxine production in B. elkanii increased ethylene evolution and decreased nodulation and competitiveness on M. atropurpureum. These results suggest that rhizobitoxine enhances nodulation and competitiveness of B. elkanii on M. atropurpureum.     

Rapid effects of indoleacetic Acid and ethylene on the growth of intact pea roots

Root auxanometers were used to determine the growth rates of individual intact primary roots accurately and quickly. The growth of pea (Pisum sativum L.) roots was inhibited by both indoleacetic acid and ethylene within 20 minutes. A supramaximal concentration of ethylene inhibited root growth less than did 5 to 20 mum indoleacetic acid, indicating that inhibition of root growth by auxin was not due only to indoleacetic acid-induced ethylene production. Inhibition of root growth was largely relieved within 60 minutes of removal of both growth regulators.     

A Mutation Altering Auxin Homeostasis and Plant Morphology in Arabidopsis

Many aspects of plant development are associated with changing concentrations of the phytohormone auxin. Several stages of root formation exhibit extreme sensitivities to exogenous auxin and are correlated with shifts in endogenous auxin concentration. In an effort to elucidate mechanisms regulating development of adventitious roots, an ethyl methanesulfonate-mutagenized M2 population of Arabidopsis was screened for mutants altered in this process. A recessive nuclear mutant, rooty (rty), displayed extreme proliferation of roots, inhibition of shoot growth, and other alterations suggesting elevated responses to auxin or ethylene. Wild-type Arabidopsis seedlings grown on auxin-containing media phenocopied rty, whereas rty seedlings were partially rescued on cytokinin-containing media. Analysis by gas chromatography-selected ion monitoring-mass spectrometry showed endogenous indole-3-acetic acid concentrations to be two to 17 times higher in rty than in the wild type. Dose-response assays with exogenous indole-3-acetic acid indicated equal sensitivities to auxin in tissues of the wild type and rty. Combining rty with mutations conferring resistance to auxin (axr1-3) or ethylene (etr1-1) suggested that root proliferation and restricted shoot growth are auxin effects, whereas other phenotypic alterations are due to ethylene. Four mutant alleles from independently mutagenized populations were identified, and the locus was mapped using morphological and restriction fragment length polymorphism markers to 3.9 centimorgans distal to marker m605 on chromosome 2. The wild-type RTY gene product may serve a critical role in regulating auxin concentrations and thereby facilitating normal plant growth and development.     

Effects of some organic solvents on ethylene evolution from young cotton bolls

The presence of promoter(s) of ethylene biosynthesis in young cotton (Gossypium hirsutum L.) fruits (bolls) was demonstrated by injection of an aqueous extract from bolls into other bolls and measurement of a 3-fold increase in rate of ethylene evolution. Injection of methionine did not affect rate of ethylene production, indicating that the promoter extracted from bolls was not methionine. Injection of the ethoxy analog of rhizobitoxine inhibited ethylene production, indicating that methionine is a precursor of ethylene in cotton bolls. Injection of organic solvents altered membrane permeability, as indicated by decreased resistance to electric current at 1,000 Hz, and stimulated ethylene evolution. The less polar solvents caused large increases in ethylene evolution, major loss of resistance, and visible evidence of membrane damage. The results support the hypothesis that membrane integrity affects rate of ethylene biosynthesis.     

Effect of Competition and Treatment with Inhibitor of Ethylene Perception on Growth and Hormone Content of Lettuce Plants

We elucidated the effect of increased planting density (single and grouped competing plants) on concentrations of auxin, abscisic acid, and cytokinins in normal lettuce plants and in those with ethylene perception inhibited by 1-methylcyclopropene (1-MCP). An attempt was made to relate the changes in hormone concentration induced by competition and inhibition of ethylene sensitivity to growth responses of lettuce planting. The results showed changes in concentrations of auxins, cytokinins, and ABA in the response of lettuce to crowding. Accumulation of ABA in shoots was likely to contribute to inhibition of transpiration of the plants grown in the presence of neighbors. This assumption was supported by the results of application of an inhibitor of ABA synthesis (fluridone and carotenoid biosynthesis herbicide) resulting in increased transpiration of grouped, but not single plants. Increased planting density led to the decline in root auxins paralleled by inhibition of root growth. This effect was likely to be due to decreased auxin transport to the roots from the shoots suggested by accumulation of auxins in the shoots and inhibition of root growth by application of the auxin transport inhibitor [N-(1-naphtyl)phtalamic acid (NPA)]. Importance of the changes in hormone concentrations was confirmed by data showing that disturbance of auxin and cytokinin distribution detected in MCP-treated plants was accompanied by corresponding modification of the growth response.

     

Effect of Galactose and Other Monosaccharides on IAA Movement in Bean Hypocotyl Segments

Galactose enhances the production of ethylene gas, and ethylene gas inhibits the movement of IAA in plant tissues. If galactose enhances ethylene production and ethylene inhibits auxin movement, then galactose should inhibit auxin movement. The above hypothesis was examined by observing the effects of d -galactose, d -inannose, d -arabinose, d -glucose, and d xylose on the uptake, presumed decarboxylation, efflux, velocity and metabolism of labeled indole-3-aectic acid in hypocotyl segments of Phaseolus vulgaris L. cv. Pinto. Galactose inhibited, arabinose and glucose enhanced, and mannose and xylose had no effect on partitioning of auxin between tissue and receptor. The reduction of auxin efflux by galactose was related to an increased presumed decarboxylation, reduced uptake and slower velocity of applied auxin. The relationship between galactose-induced growth effects, ethylene production, and auxin migration are discussed.     

Detailed quantitative analysis of architectural traits of basal roots of young seedlings of bean in response to auxin and ethylene

Vertical placement of roots within the soil determines their efficiency of acquisition of heterogeneous belowground resources. This study quantifies the architectural traits of seedling basal roots of bean (Phaseolus vulgaris), and shows that the distribution of root tips at different depths results from a combined effect of both basal root growth angle (BRGA) and root length. Based on emergence locations, the basal roots are classified in three zones, upper, middle, and lower, with each zone having distinct architectural traits. The genotypes characterized as shallow on BRGA alone produced basal roots with higher BRGA, greater length, and more vertically distributed roots than deep genotypes, thereby establishing root depth as a robust measure of root architecture. Although endogenous indole-3-acetic acid (IAA) levels were similar in all genotypes, IAA and 1-N-naphthylphthalamic acid treatments showed different root growth responses to auxin because shallow and deep genotypes tended to have optimal and supraoptimal auxin levels, respectively, for root growth in controls. While IAA increased ethylene production, ethylene also increased IAA content. Although differences in acropetal IAA transport to roots of different zones can account for some of the differences in auxin responsiveness among roots of different emergence positions, this study shows that mutually dependent ethylene-auxin interplay regulates BRGA and root growth differently in different genotypes. Root length inhibition by auxin was reversed by an ethylene synthesis inhibitor. However, IAA caused smaller BRGA in deep genotypes, but not in shallow genotypes, which only responded to IAA in the presence of an ethylene inhibitor.