Endogenous ethylene and reversing methyl jasmonate inhibition of Amaranthus caudatus seed germination by benzyladenine or gibberellin

BA at 10^–5 M, GA₃ at 3×10^–4 M or GA₄₊₇ at 3×10^–5 M partially or largely reversed the inhibition of Amaranthus caudatus seed germination due to JA-Me. BA or GA₃ did not affect ethylene production and ACC oxidase activity in vivo in the presence of JA-Me before radicle protrusion. However, both increased ethylene production after 72 h of incubation, when the reversal of the JA-Me inhibition of seed germination was observed. AVG at 3×10^–4 M decreased ethylene production when it was applied simultaneously with BA and JA-Me or GA₃ and JA-Me, but it had no effect on seed germination. NBD almost completely reversed the stimulatory effect of BA, GA₃ or GA₄₊₇ on the germination of seeds in the presence of JA-Me. Exogenous ethylene reversed the inhibitory effect of NBD. The results indicate that action of endogenous ethylene is involved in the response of JA-Me inhibited seeds to BA or GA_s.     

Ameliorative effect of brassinosteroid and ethylene on germination of cucumber seeds in the presence of sodium chloride

Brassinosteroids are a class of plant polyhydroxysteroids with a diverse of functions in plant growth and development, while ethylene is a gaseous hormone involved in regulation of numerous physiological processes. To evaluate the roles of BR and ethylene in seed germination under conditions of salt stress, effects of 24-Epibrassinolide (EBR) and 1-aminocyclopropane-1-carboxylic acid (ACC) on seed germination of cucumber (Cucumis sativus) seeds in the presence of 250 mM NaCl were investigated. Seed germination was significantly inhibited by the presence of NaCl in the incubation medium, and the inhibitory effect was significantly alleviated by addition of EBR and ACC to the incubation medium containing NaCl. There was an increase in ethylene evolution during seed germination and this increase was suppressed by salt stress. The reduction in ethylene evolution from imbibed seeds by salt stress was attenuated by EBR. Salt stress inhibited ACC oxidase (ACO) activity and EBR reversed the salt stress-induced decrease in ACO activity. Salt stress reduced expression of gene encoding ACO (CsACO2), and EBR reversed the salt stress-induced down-regulation of CsACO2. The alleviative effect of EBR on seed germination in the presence of NaCl was diminished by antagonist of ethylene synthesis, aminoethoxyvinylglycine. These results indicate that both ethylene and BR are likely to be associated with suppression of seed germination under salt stress and that the mitigating effect of BR on salt stress-induced inhibition of seed germination may occur through its interaction with ethylene synthesis.     

Polyamines and ethylene in relation to adventitious root formation in Prunus avium shoot cultures

An attempt was made to identify some of the hormonal factors that control adventitious root formation in our Prunus avium micropropagation system in order to improve rooting in difficult-to-root genotypes. Changes in endogenous contents of free polyamines were determined at intervals during auxin-induced rooting of shoot cultures. Accumulation of putrescine and spermidine peaked between days 9 and 11. Spermine was only present in traces, Exogenously supplied putrescine or spermine (50-500 μM), in the presence of optimal or suboptimal levels of indolebutyric acid (IBA), had no effect on rooting percentage or root density, except for spermine at 500 μM. At this external concentration spermine caused a substantial accumulation in both free spermine and putrescine. The use of several inhibitors of polyamine biosynthesis, namely α-difluoromethylornithine (DFMO), α-difluoromethylarginine (DFMA), dicyclohexylammonium sulphate (DCHA) and methylglyoxal-bis-guanyl-hydrazone (MGBG) alone or in combination in the 0.1 to 5 μM range, resulted in an inhibition of rooting that was partially reversed by the addition of the corresponding polyamine. Cellular polyamine levels were significantly reduced by DFMO and DFMA but not by DCHA and MGBG, Labeled putrescine incorporation into spermidine increased somewhat in the presence of the ethylene synthesis inhibitor aminoethoxyvinylglycine (AVG). A system based on [3,4-¹⁴C]methionine incorporation was used to measure ethylene synthesis by the in vitro cultured shoots. Label incorporation was drastically reduced by 10 μM AVG and increased 3.5-fold in the presence of 50 μM IBA with respect to controls (no IBA). Labeled methionine incorporation into spermidine increased to some extent when ethylene synthesis was inhibited by AVG. Adding the ethylene precursor 1-aminocyclopropane-l-carboxylic acid (ACC) to the rooting medium significantly inhibited rooting percentage; AVG caused the formation of a greater number of roots per shoot but delayed their growth. Supplying the shoots with both compounds resulted in an intermediate rooting response, in which both rooting percentage and root density were affected. These results indicate that polyamines may play a significant role at least in some stages of root formation. The polyamine and ethylene biosynthetic pathways seem to be competitive but under our conditions, the enhancement of one pathway when the other was inhibited, was not dramatic. Although IBA promoted ethylene synthesis, AVG, which drastically reduced it, also promoted root formation. Thus, the auxin effect on root induction cannot be directly related to its ability to enhance ethylene synthesis.     

The effect of ethylene, octanoic acid and a plant-derived smoke extract on the germination of light-sensitive lettuce seeds

The effects of an aqueous plant-derived smoke extract, octanoic acid and ethylene on germination of light-sensitive Grand Rapids lettuce seeds were investigated. The smoke extract brought about a concentration dependent increase in germination and a complete inhibition of germination at high concentrations. Octanoic acid could not induce germination. Ethylene at concentrations over 5 L L^–1 increased lettuce seed germination, but not to the same degree as smoke. Aqueous smoke in combination with ethylene showed a synergistic effect on germination at suboptimal smoke concentrations. At high smoke concentrations the effect of ethylene was almost completely inhibited. Octanoic acid in combination with ethylene brought about a higher level of germination than with ethylene alone, but only at the highest concentration of octanoic acid tested (1 mM). Standardized hexane and dichloromethane-partitioned smoke extracts and octanoic acid were subjected to TLC separation. The R _f -fractions in the smoke lanes showing activity in the lettuce seed bioassay did not correspond to the R _f -value of octanoic acid. As aqueous smoke can withstand autoclaving and can be separated by TLC and HPLC without loosing activity it is unlikely that the activity of aqueous smoke is linked to ethylene. It thus appears that the active compound in smoke is neither octanoic acid nor ethylene.Abbreviations TLC thin layer chromatography - HPLC High performance liquid chromatography     

Photophysiology of turion germination inSpirodela polyrhiza (L.)Schleiden. The cause of germination inhibition by overcrowding

Red-light-induced (via phytochrome) germination decreased with increasing numbers of turions per germination flask (overcrowding). Three hypotheses concerning the mechanism of this germination inhibition were tested, related to abscisic acid, ethylene, and oxygen deficiency: (i) Although abscisic acid is a powerful inhibitor of turion germination it had to be excluded as a cause, because abscisic acid was not secreted from turions into the nutrient solution, (ii) Ethylene (ethrel) strongly inhibited growth of newly formed sprouts, but germination response itself was not inhibited, (iii) Germination inhibition did not appear if short light pulses were substituted by continuous irradiation. It reappeared in the presence of the photosynthesis inhibitor 3-(3, 4-dichlorophenyl)-l, 1-dimethylurea, but it was not observed in aerated nutrient solutions, or when Petri dishes instead of Erlenmeyer flasks were used. Decreased oxygen concentrations in the nutrient solution were produced by turion respiration. Consequently, anaerobiosis within the nutrient solution caused by turion respiration was the reason for germination inhibition by overcrowding.     

Inhibitory Effects of Methyl Jasmonate on the Germination and Ethylene Production in Cocklebur Seeds

Methyl jasmonate (JA-Me) inhibited the germination of cocklebur (Xanthium pennsylvanicum Wallr.) seeds. The inhibition of the germination of cocklebur seeds treated with JA-Me at concentrations less than 300 μm was nullified by ethylene applied exogenously, although the inhibitory effect of 1,000 μm JA-Me was not recovered completely even by high concentrations of ethylene (10,000 μL/liter). JA-Me inhibited ethylene production before seed germination. The level of 1-aminocyclopropane-1-carboxylic acid (ACC) in the cotyledonary tissues treated with JA-Me decreased but not the level of 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC). JA-Me inhibited the conversion of ACC to ethylene in the tissues. These results suggested that JA-Me inhibits ethylene production by prevention of ACC oxidation in addition to ACC synthesis. We believe that the inhibition of ethylene production by JA-Me results in the retardation of the germination of cocklebur seeds. Received June 4, 1997; accepted October 23, 1997     

GERMINATION OF LIGHT-INHIBITED SEED OF NEMOPHILA INSIGNIS

Germination of Nemophila insignis seed is inhibited by light over a wide range of temperatures. At low temperatures the light intensity required for inhibition is higher. At 25 C there is little germination (in darkness as well as in light); at 27.5 C germination is inhibited altogether. Virtually complete germination in light is obtained when the endosperm directly covering the radicle is removed. This operation also improves germination in darkness at 25 C. Mechanical scarification performed elsewhere in the seed is without effect. As with Phacelia tanacetifolia, Nemophila seed apparently fails to germinate in light because the endosperm restrains the expansive growth of the embryo. The embryo of dark-imbibed seed develops a force which enables it to overcome the physical resistance of the endosperm. Light inhibits the process which leads to generation of “expansive force.” Gibberellic acid at 5 × 10^–4 m stimulates germination in light and in the dark. Abscisic acid at 10^-4 m inhibits germination; at 10^-6 m it inhibits only root growth. The inhibition of germination or root growth caused by abscisic acid cannot be reversed by gibberellic acid. Eighty per cent oxygen under certain conditions promotes germination. The rate of O₂ uptake is enhanced in oxygen-enriched atmosphere; however, there is no corresponding increase in the rate of CO₂ output. Thus high oxygen tension enhances an oxidative process other than respiration. This reaction is favorable to seed germination.     

Effects of Dormancy Regulating Chemicals on Innate and Salinity Induced Dormancy in the Invasive <Emphasis Type="Italic">Prosopis juliflora</Emphasis> (Sw.) DC. Shrub

The effect of different types and concentrations of dormancy regulating chemicals (DRCs) on innate and induced dormancy was evaluated under optimal germination conditions in the invasive Prosopis juliflora shrub. Lower concentrations of gibberellic acid (0.3 mM) and kinetin (0.05 mM) were more effective in enhancing germination % and rate at higher concentrations of NaCl, but the reverse was true for thiourea. None of the DRCs alleviated innate dormancy of P. juliflora. Germination % and rate decreased as salinity increased. Percent final germination of non-treated seeds was significantly reduced at 500 mM NaCl and virtually inhibited in 600 mM NaCl. Germination reduction in 500 mM NaCl was not alleviated by any of the DRCs, but inhibition induced at 600 mM NaCl was partially alleviated by all the DRCs. Gibberellic acid had a significantly greater effect than kinetin in alleviating germination inhibition. For restoration of saline soil through the use of P. juliflora, results suggest using DRCs, particularly gibberellic acid and thiourea, as a preseeding treatment can overcome the problem of reduced germination.     

Effect of water stress on seed germination and seedling growth inOryza sativa L.

The effect of water stress induced by application of polyethylene glycol 6000 during seed germination and seedling growth ofOryza sativa L. cv. IAC 165 was analysed. The seed germination was inhibited by the decrease in the water potential of the medium, the inhibition being greater under white light than under continuous darkness. When the seedling was submitted to water stress (-0.51 MPa) white light inhibited growth of root, coleoptile-and leaf, while under no stress conditions white light caused increase in growth of root and leaf and only inhibition of coleoptile growth.     

Inhibition of Amaranthus caudatus seed germination by polyethylene glycol-6000 and abscisic acid and its reversal by ethephon or 1-aminocyclopropane-1-carboxylic acid

The effect of polyethylene glycol (PEG-6000), abscisic acid (ABA), 2-chloroethylphosphonic acid (ethephon) and 1-aminocyclopropane-1-carboxylic acid (ACC) on the germination of Amaranthus caudatus L. seeds was examined. Both PEG-6000 and ABA inhibited the rate and percentage of seed germination. ABA potentiated the effect of PEG. Ethephon was highly effective in reversing the inhibitory effect of PEG and ABA or combinations of both. ACC relieved inhibition by ABA and the combined effect of ABA and PEG. Aminoethoxyvinylglycine (AVG) increased the inhibition of seed germination caused by ABA. The inhibition of seed germination by ABA seems to be related more or less to ethylene biosynthesis or is associated with a change of tissue sensitivity to ethylene. The possibility of ethylene control of water uptake by seeds is also considered.     

ON THE MECHANISMS OF LIGHT-INDUCED GERMINATION INHIBITION OF PHACELIA TANACETIFOLLV

Germination of seed of Phacelia tanacetifolia is inhibited by several mechanisms. In addition to physical restraints imposed by the seed coats, the seed contains a water-soluble inhibitor which is independent of light or temperature for its activity. Available evidence also points to the presence of 1 or more light-activated inhibitors which are not easily leached from the seed. The blue-light-activated inhibition can be negated by high oxygen tensions or mechanical abrasion of the micropylar end of the seed. The suppression of germination by far-red or red light can be negated by abrasion but is only partially reversed by oxygen. Combinations of abrasion and high oxygen tensions negate both light-induced and temperature-induced inhibitions of germination.     

Effect of malformin on phytochrome- and Ethrel-mediated responses

On etiolated cuttings of Phaseolus vulgaris malformin inhibitedseveral phytochromemediated responses. This included hypocotylhook-opening, leaf expansion, and inhibition of stem elongation.Malformin also inhibited anthocyanin synthesis by sorghum, buthad no effect upon lettuce seed germination in the light ordark. Malformin alleviated Ethrel-induced hook-retention, inhibitionof stem elongation, and root curvatures, but not Ethrel-inducedstimulation of lettuce seed germination in the dark. (Received February 19, 1975; )     

FACTORS INFLUENCING SEED GERMINATION IN SALICORNIA PACIFICA VAR. UTAHENSIS

The halophyte, Salicornia pacifica var. utahensis (Tiderstorm) Munz produces seed under high salinity conditions, and deposits its seed on saline soil. Experiments were conducted to determine the effect of salinity, temperature and growth regulators on germination. Results indicate that the seeds can germinate at very high salt concentration (5% NaCl). Germination was sensitive to the changes in temperature regimes. At higher 30–20 C, light-dark sequence, no germination occurred at 3, 4 and 5% NaCl treatments. On the other hand, 30% germination did occur at 5% NaCl treatment at a temperature regime of 15–5 C. These seeds required light for germination. Only 50% germination occurred in the non-saline control in the dark and the addition of NaCl further reduced germination. The GA₃ partially alleviated the inhibitory effect of NaCl and darkness. Kinetin did not promote germination.     

Inhibition of Lettuce Seed Germination and Root Elongation by Derivatives of Auxin and Reversal by Derivatives of Cycocel

Lettuce seed germination or lettuce root elongation after germination in water was inhibited by 5.7 × 10^-6M indoleacetic acid (IAA) and designated other IAA derivatives. These IAA-inhibited growth responses were reversed by 10^-5 to 10^-6M Cycocel, (2-chloroethyl) trimethylammonium chloride, which alone was without effect. Only those Cycocel analogs, which have previously been shown to be active as plant growth retardants, were effective in reversing IAA inhibition of germination or root elongation. These results are consistent with the concept that Cycocel at low concentrations acts as an auxin antagonist. However. Cycocel did not reverse the inhibitory effects from indole-3-propionic acid or indole-3-butyric acid.     

Regulation of α-amylase activity in Amaranthus caudatus seeds by methyl jasmonate, gibberellin A3, benzyladenine and ethylene

Methyl jasmonate (JA-Me) at 10^–3 Minhibited Amaranthus caudatus seed germination anddecreased -amylase activity. Exogenous gibberellin A₃(GA₃) and ethylene, but not benzyladenine (BA), increased activity ofthe enzyme in the presence of JA-Me, with ethylene being the most effective. Theinhibitor of ethylene action, 2,5-norbornadiene (NBD) inhibited seed germinationand decreased -amylase activity. The inhibitory action of JA-Me onAmaranthus caudatus seed germination is associated with theinhibition of -amylase activity. Exogenous GA₃ and ethylenecontrol both -amylase activity and seed germination in the presence of JA-Me.     

Ethylene Production and 1-Aminocyclopropane-1-Carboxylic Acid Conjugation in Thermoinhibited Cicer arietinum L. Seeds

The effect of supraoptimal temperatures (30°C, 35°C) on germination and ethylene production of Cicer arietinum (chick-pea) seeds was measured. Compared with a 25°C control, these temperatures inhibited both germination and ethylene production. The effect of supraoptimal temperatures could be alleviated by treating the seeds with ethylene. It was concluded that one effect of high temperature on germination was due to its negative effect on ethylene production. This inhibitory effect of high temperature was due to increased conjugation of 1-aminocyclopropane-1-carboxylic acid to 1-(malonylamino)cyclopropane-1-carboxylic acid and to an inhibition of ethylene-forming enzyme activity.     

In vitro and in vivo antagonism of biocontrol agents against Colletotrichum lindemuthianum causing bean anthracnose

Bean anthracnose caused by Colletotrichum lindemuthianum is a serious seed borne disease. For devising an effective management strategy, the efficacy of different bioagents, viz. Trichoderma viride, Trichoderma harzianum, Trichoderma hamatum and Gliocladium virens conducted under in vitro and in vivo conditions revealed maximum inhibition of mycelial growth in dual culture (59.48%) and inverted plate (55.98%) with T. viride. All the bioagents overgrew the pathogen and the principal mechanism of mycoparisitism observed was coiling, brusting and disintegration of pathogen hyphae. Culture filtrate from T. viride was found best as it completely inhibited radial growth at 25 and 50% concentration and reduced the spore germination of test fungus significantly. However, lower concentrations of culture filtrate from all bioagents showed little effect on spore germination. Seed application of bioagents was found better as compared to soil application. A maximum increase in seed germination and inhibition of seed borne infection was observed with T. viride followed by T. harzianum under pot culture conditions. T. viride has the maximum potentiality to suppress the spore germination, mycelial growth, seed borne infection of C. lindemuthianum and increased seed germination when compared with the other biocontrol agents.     

Action of ferric and aluminium ions on the dormancy breakage of <Emphasis Type="Italic">Stylosanthes humilis</Emphasis> seeds

Dormancy of scarified seeds of Stylosanthes humilis was broken by acidic Al³⁺ and Fe³⁺ solutions. Fe⁺³-stimulated seeds exhibited a high activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and produced great amounts of ethylene, which showed correlated with the germination process. In addition, specific inhibitors of ethylene biosynthesis and action largely depressed the Fe³⁺-stimulated germination, leading to the conclusion that the ion broke dormancy by triggering ethylene production by the seeds. By contrast, inhibitors of ethylene biosynthesis and action did not impair germination of Al³⁺-stimulated dormant seeds. Moreover, ethylene production and activity of ACC oxidase of Al³⁺-treated seeds was substantially decreased by inhibitors of ethylene biosynthesis, but germination kept large. Together these data suggest that ethylene biosynthesis was not required in the chain of events triggered by Al³⁺ leading to dormancy breakage. Methyl viologen (MV), a reactive oxygen species-generating compound, broke dormancy of seeds to the same extent as Al³⁺ did. Germination of both Al³⁺- and MV-stimulated dormant seeds was inhibited by sodium selenate, an antioxidant compound; selenate, however had no effect on germination of Fe³⁺-stimulated seeds. Together these data indicate that the mechanisms underlying the germination of Al³⁺- and Fe³⁺-treated seeds are not the same.     

Cytokinin Reversal of Abscisic Acid Inhibition of Growth and α-Amylase Synthesis in Barley Seed

The effect of cytokinin, kinetin, on abscisic acid (dormin) inhibition of α-amylase synthesis and growth in intact barley seed was investigated. Abscisic acid at 5 × 10^?5M nearly completely inhibited growth response and α-amylase synthesis in barley seed. Kinetin reversed to a large extent abscisic acid inhibition of α-aniylase synthesis and coleoptile growth. The response curves of α-amylase synthesis and coleoptile growth in presence of a fixed amount of abscisic acid (6 × l0^?6M) and increasing concentrations of kinetin (from 5 × l0^?7M to 5 × 10^?5 M) showed remarkable similarity. Kinetin and abscisic acid caused synergistic inhibition of root growth. Gibberellic acid was far less effective than kinetin in reversing abscisic acid inhibition of α-amylase synthesis and coleoptile growth. A combination of kinetin and gibberellic acid caused nearly complete reversal of abscisic acid inhibition of α-amylase synthesis but not the abscisic acid inhibition of growth. The results suggest that factors controlling α-amylase synthesis may not have a dominant role in all growth responses of the seed. Kinetin possibly acts by removing the abscisic acid inhibition of enzyme specific sites thereby allowing gibberellic acid to function to produce α-amylase.     

Germination and Dormancy of Reed Canary-Grass Seeds (Phalaris arundinacea)

Effects of various chemical and physical factors on the germination of several seed lots of reed canary-grass (Phalaris arundinacea L.) have been studied. Germination at the optimum constant temperatures of 24 to 27°C was significantly stimulated by the following treatments: moist chilling in light, red light given during the first 3 days of imbibition, three 2-h periods at 12°C given during the second day of imbibition, ethylene, increased oxygen tension and soaking in aerated water for 4 days. Dry storage at 20–30°C had no effect on the germination ability of the seeds. No significant quantities of germination inhibitors were found either in water or methanol extracts of seed dispersal units. By comparing three cultivars with various degrees of seed dormancy, respiration measurements showed that there was a significant positive correlation between oxygen uptake prior to visible germination and germination capacity. Similarly, germination-stimulating treatment significantly enhanced oxygen uptake prior to visible germination.