Evidence that ethylene, light and abscisic acid interact to inhibit germination in the recalcitrant seeds of Quercus robur L.

The effects of ethylene, light and abscisic acid (ABA) on thegermination of prematurely-harvested and mature Q. robur seedswere studied. There was evidence of active biosynthesis of ethylene,but not of ABA in seeds from both harvests. Addition of ABA,the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid(ACC) or the ethylene-releasing compound Ethephon (E) to germinationmedia and exposure to light individually inhibited germinationand their effects were additive. The ethylene biosynthesis inhibitoraminoethoxyvinylglycine (AVG) tended to promote germinationof premature seeds only. Key words: Quercus robur, germination, ethylene, light, abscisic acid, recalcitrant seed     

Interactions between abscisic acid and ethylene signaling cascades

We screened for mutations that either enhanced or suppressed the abscisic acid (ABA)-resistant seed germination phenotype of the Arabidopsis abi1-1 mutant. Alleles of the constitutive ethylene response mutant ctr1 and ethylene-insensitive mutant ein2 were recovered as enhancer and suppressor mutations, respectively. Using these and other ethylene response mutants, we showed that the ethylene signaling cascade defined by the ETR1, CTR1, and EIN2 genes inhibits ABA signaling in seeds. Furthermore, epistasis analysis between ethylene- and ABA-insensitive mutations indicated that endogenous ethylene promotes seed germination by decreasing sensitivity to endogenous ABA. In marked contrast to the situation in seeds, ein2 and etr1-1 roots were resistant to both ABA and ethylene. Our data indicate that ABA inhibition of root growth requires a functional ethylene signaling cascade, although this inhibition is apparently not mediated by an increase in ethylene biosynthesis. These results are discussed in the context of the other hormonal regulations controlling seed germination and root growth.     

ACC conversion to ethylene by sunflower seeds in relation to maturation,germination and thermodormancy

Conversion of exogenous 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene was studied in sunflower (Helianthus annuus L., cv. Mirasol) seeds in relation to germinability. Ethylene production from ACC decreased during seed maturation, and non-dormant mature seeds were practically unable to synthesize ethylene until germination and growth occurred, indicating that ethylene forming enzyme (EFE) activity developed during tissue imbibition and growth. ACC conversion to ethylene was reduced by the presence of pericarp, and in young seedlings it was less in cotyledons than in growing axes.ACC conversion to ethylene by cotyledons from young seedlings was optimal at c. 30°C, and was strongly inhibited at 45°C. Pretreatment of imbibed seeds at high temperature (45°C) induced a thermodormancy and a progressive decrease in EFE activity.Abscisic acid and methyl-jasmonate, two growth regulators which inhibit seed germination and seedling growth, and cycloheximide were also shown to inhibit ACC conversion to ethylene by cotyledons of 3-day-old seedlings and by inbibed seeds.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - CH cycloheximide - EFE ethylene forming enzyme - IAA indole-3-acetic acid - Me-Ja methyl-jasmonate     

Ethylene biosynthesis in Amaranthus caudatus seeds in response to methyl jasmonate

Methyl jasmonate (JA-Me) at 10^–3 M completely inhibited Amaranthus caudatus seed germination. Exogenous ethylene could totally reverse this inhibition. The inhibitor of ethylene action, 2,5-norbornadiene (NBD), increased the sensitivity of seeds to JA-Me. Methyl jasmonate inhibited ethylene production and also decreased both 1-aminocyclopropane-1-carboxylic acid (ACC) and malonyl ACC (MACC) content. Likewise, ACC oxidase activity in vivo was decreased by jasmonate. Similarly ACC oxidase activity in vitro isolated from seeds incubated in the presence of JA-Me was lower than that isolated from untreated seeds.The inhibitory JA-Me action on seed germination seems to be mainly associated with the inhibition of ethylene biosynthesis. Both inhibition of ACC synthase and ACC oxidase activity and/or synthesis can be involved.     

Ethylene promotes ethylene biosynthesis during pea seed germination by positive feedback regulation of 1-aminocyclo-propane-1-carboxylic acid oxidase

 Increased ethylene evolution accompanies seed germination of many species including Pisum sativum L., but only a little is known about the regulation of the ethylene biosynthetic pathway in different seed tissues. Biosynthesis of the direct ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), the expression of ACC oxidase (ACO), and ethylene production were investigated in the cotyledons and embryonic axis of germinating pea seeds. An early onset and sequential induction of ACC biosynthesis, accumulation of Ps-ACO1 mRNA and of ACO activity, and ethylene production were localized almost exclusively in the embryonic axis. Maximal levels of ACC, Ps-ACO1 mRNA, ACO enzyme activity and ethylene evolution were found when radicle emergence was just complete. Treatment of germinating seeds with ethylene alone or in combination with the inhibitor of ethylene action 2,5-norbornadiene showed that endogenous ethylene regulates its own biosynthesis through a positive feedback loop that enhances ACO expression. Accumulation of Ps-ACO1 mRNA and of ACO enzyme activity in the embryonic axis during the late phase of germination required ethylene, whereas Ps-ACS1 mRNA levels and overall ACC contents were not induced by ethylene treatment. Ethylene did not induce ACO in the embryonic axis during the early phase of germination. Ethylene-independent signalling pathways regulate the spatial and temporal pattern of ethylene biosynthesis, whereas the ethylene signalling pathway regulates high-level ACO expression in the embryonic axis, and thereby enhances ethylene evolution during seed germination. Received: 28 September 1999 / Accepted: 27 December 1999     

Enhancement of ethylene biosynthesis and germination by cytokinins and 1-aminocyclopropane-l-carboxylic acid in Striga asiatica seeds

Germination of witchweed ( Striga asiatica [L.] Kuntze), an important parasitic weed on several poaceous crops, is stimulated by several synthetic and natural compounds. We investigated the role of ethylene biosynthesis and action in cytokinin-induced germination. Conditioned Striga seeds treated with distilled water, 1-aminocyclopro-pane-1-carboxylic acid (ACC) or the cytokinins thidiazuron (TDZ), trans zeatin (TZ), benzyladenine (BA) and kinetin (KIN) produced little ethylene. Treatments with cytokinin-ACC combinations enhanced ethylene production. The relative order of activity of the cytokinins in elicitation of the phytohormone was TDZ > TZ > BA > KIN. Germination in response to distilled water and ACC treatments was negligible. Induction of germination by cytokinins varied from low (0%) to moderate (52%). Seeds treated with cytokinin-ACC combinations displayed high rates of germination. The observed germination was positively correlated (γ= 0. 8 and 0. 9) with ethylene production. Germination was reduced by silver thiosulphate (STS) and CoCl₂, inhibitors of ethylene action and ACC oxidase, respectively. Aminoethoxyvi-nylglycine (AVG), an ACC-synthase inhibitor, reduced TDZ-induced Striga germination. However, the inhibitory effect of AVG was overcome by addition of ACC. The results are consistent with a model in which Striga germination and embryo growth are limited by low capacity of the seeds to oxidize ACC. The cytokinins promote ACC conversion into ethylene and consequent Striga germination by enhancing ACC oxidase activity and/or synthesis.     

The 7B-1 mutant in tomato shows blue-light-specific resistance to osmotic stress and abscisic acid

Germination of wild-type (WT) tomato ( Lycopersicon esculentum Mill.) seed is inhibited by mannitol (100-140 mM) in light, but not in darkness, suggesting that light amplifies the responsiveness of the seed to osmotic stress (M. Fellner, V.K. Sawhney (2001) Theor Appl Genet 102:215-221). Here we report that white light (W) and especially blue light (B) strongly enhance the mannitol-induced inhibition of seed germination, and that the effect of red light (R) is weak or nil. The inhibitory effect of mannitol could be completely overcome by fluridone, an inhibitor of abscisic acid (ABA) biosynthesis, indicating that mannitol inhibits seed germination via ABA accumulation in seeds. The inhibition of WT seed germination by exogenous ABA was also amplified by W or B, but not by R. In a recessive, ABA-overproducing, 7B-1 mutant of tomato, seed germination and hypocotyl growth were resistant to inhibition by mannitol or exogenous ABA, both in W or B. Experiments with fluridone suggested that inhibition of hypocotyl growth by W or B is also partially via ABA accumulation. De-etiolation in the mutant was especially less in B compared to the WT, and there was no difference in hypocotyl growth between the two genotypes in R. Our data suggest that B amplifies the responsiveness of tomato seeds and hypocotyls to mannitol and ABA, and that W- or B-specific resistance of the 7B-1 mutant to osmotic stress or ABA is a consequence of a defect in B perception or signal transduction.     

干旱胁迫与不同储藏时间对杠柳种子萌发的影响

为了阐明干旱胁迫与不同储藏时间对杠柳种子萌发的影响机理,在室内采用不同浓度的聚乙二醇（PEG）模拟干旱胁迫条件,对3种不同储藏时间的杠柳种子进行了萌发期对比试验,观测了种子的总萌发率、发芽势、活力指数和胚根的生长情况,分析了抗旱特性,结果表明：不同程度的干旱胁迫与储藏时间对杠柳种子萌发均有一定的影响,当PEG-6000浓度为5%,对储藏1年杠柳种子的萌芽率和发芽势起促进作用,对储藏2年种子萌芽率轻度抑制,对储藏3年种子抑制影响较大,当PEG-6000浓度>5%,3种种子的萌芽率和发芽势均呈下降趋势,但当胁迫浓度增大到25%时,种子基本没有萌发。在不同的水分胁迫处理中,3种种子的活力指数和胚根生长速度一直呈下降趋势,尤其3年种子的下降幅度最大。不同储藏时间杠柳种子对干旱胁迫的适应性有差异,储藏1~2年的杠柳种子在PEG-6000浓度为5%~15%中萌发效果较好,且储藏1年种子好于2年种子,3年种子较差。3种种子的耐旱能力由强到弱依次为：1年种子>2年种子>3年种子。     

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     

Control of macaw palm seed germination by the gibberellin/abscisic acid balance

The hormonal mechanisms involved in palm seed germination are not fully understood. To better understand how germination is regulated in Arecaceae, we used macaw palm (Acrocomia aculeata (Jacq.) Lodd. Ex Mart.) seed as a model. Endogenous hormone concentrations, tocopherol and tocotrienol and lipid peroxidation during germination were studied separately in the embryo and endosperm. Evaluations were performed in dry (D), imbibed (I), germinated (G) and non‐germinated (NG) seeds treated (+GA₃) or not treated (control) with gibberellins (GA). With GA₃ treatment, seeds germinated faster and to a higher percentage than control seeds. The +GA₃ treatment increased total bioactive GA in the embryo during germination relative to the control. Abscisic acid (ABA) concentrations decreased gradually from D to G in both tissues. Embryos of G seeds had a lower ABA content than NG seeds in both treatments. The GA/ABA ratio in the embryo was significantly higher in G than NG seeds. The +GA₃ treatment did not significantly affect the GA/ABA ratio in either treatment. Cytokinin content increased from dry to germinated seeds. Jasmonic acid (JA) increased and 1‐aminocyclopropane‐1‐carboylic acid (ACC) decreased after imbibition. In addition, α‐tocopherol and α‐tocotrienol decreased, while lipid peroxidation increased in the embryo during germination. We conclude that germination in macaw palm seed involves reductions in ABA content and, consequently, increased GA/ABA in the embryo. Furthermore, the imbibition process generates oxidative stress (as observed by changes in vitamin E and MDA).     

Ethylene in seed dormancy and germination

The role of ethylene in the release of primary and secondary dormancy and the germination of non-dormant seeds under normal and stressed conditions is considered. In many species, exogenous ethylene, or ethephon – an ethylene-releasing compound - stimulates seed germination that may be inhibited because of embryo or coat dormancy, adverse environmental conditions or inhibitors (e.g. abscisic acid, jasmonate). Ethylene can either act alone, or synergistically or additively with other factors. The immediate precursor of ethylene biosynthesis, 1-aminocyclopropane-1-carboxylic acid (ACC), may also improve seed germination, but usually less effectively. Dormant or non-dormant inhibited seeds have a lower ethylene production ability, and ACC and ACC oxidase activity than non-dormant, uninhibited seeds. Aminoethoxyvinyl-glycine (AVG) partially or markedly inhibits ethylene biosynthesis in dormant or non-dormant seeds, but does not affect seed germination. Ethylene binding is required in seeds of many species for dormancy release or germination under optimal or adverse conditions. There are examples where induction of seed germination by some stimulators requires ethylene action. However, the mechanism of ethylene action is almost unknown.
The evidence presented here shows that ethylene performs a relatively vital role in dormancy release and seed germination of most plant species studied.     

Hormonal regulation of tomato seed germination at a supraoptimal temperature

Germination of tomato cv. New Yorker seed is inhibited at 35°C. This thermoinhibition was partially counteracted by application of GA₄₊₇ alone, the compound applied in combination with ACC or ethephon markedly enhancing the process. The latter compound alone was not able to induce germination at 35 °C. Thermoinhibition of seeds at 35 °C was also counteracted by fluridone, an inhibitor of ABA biosynthesis. At 25 °C, an optimal temperature, ABA inhibited germination of New Yorker seeds. Although another known growth inhibitor MeJA, when applied at an optimal temperature (25 °C), had also a slightly inhibitory effect on germination of those seeds and clearly delayed the process, inhibitors of its biosynthetic pathway (ibuprofen, indoprofen, antypiryne and salicylic acid) did not remove thermoinhibition at 35 °C. An increase in endo-β-mannanase activity after 24 hours of incubation at 35 °C was observed in the seeds incubated in the presence of gibberellins, ACC, ethephon, fluridone used alone and in combinations, but it was not clearly correlated with the effects of these compounds on alleviation of seed germination. However, fluridone present in the same incubation medium at 35 °C with ABA was able to counteract the inhibitory effect of ABA on endo-β-mannanase activity. The results of our study suggest that gibberellins, ethylene (produced from ACC or ethephon) and ABA, but not jasmonates, regulate tomato seed germination at supraoptimal temperatures. Alleviation of thermoinhibition of New Yorker seed germination by plant growth regulators and fluridone is partially associated with their controlling endo-β-mannanase activity.     

The effects of abscisic acid and methyl jasmonate on 1-aminocyclopropane 1-carboxylic acid conversion to ethylene in hypocotyl segments of sunflower seedlings,and their control by calcium and calmodulin

The conversion of 1-aminocyclopropane 1-carboxylic acid (ACC) to ethylene by hypocotyl segments of sunflower (Helianthus annuus L.) seedlings was inhibited by abscisic acid (ABA) and methyl jasmonate (Me-Ja), and this inhibitory effect increased with increasing concentration of both growth regulators. On the contrary, CaCl, enhanced ACC conversion to ethylene at the concentrations of 10^-4 M and 5 x 10^-4 M, however lower and higher concentrations had no significant action. CaCl, (5 x 10^-4M) seemed to magnify the inhibition of the reaction induced by ABA, whereas it reduced (5 x 10^-4M) and even abolished (10^-3M) the inhibitory action of Me-Ja. The results obtained with a Ca²⁺ chelator (EGTA), a Ca²⁺ channel blocker (nifedipine) and calmodulin antagonists (W7 and TFP), given in association with ABA or Me-Ja, suggested that calcium was involved in the inhibition of ACC conversion to ethylene by ABA and Me-Ja through an interaction with calmodulin. However, the mechanism of action of the two growth regulators seemed to be different, since all treatments which resulted in a decrease in cytosolic Ca²⁺ concentration or in calmodulin action induced a decrease in the effect of ABA and an increase in the effect of Me-Ja.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane 1-carboxylic acid - EFE ethylene for enzyme - EGTA ethylene glycol-bis-2-aminoethyl tetraacetic acid - Me-Ja methyl jasmonate - NIF nifedipine - TFP trifluoperazine dihydrochloride - W7 N-(6-aminohexyl)5-chloro-l-naphthalenesulfonamide hydrochloride     

粗茎秦艽种子粗提物与破眠方法研究

为了探究影响粗茎秦艽（Gentiana crassicaulis Duthie ex Burk.）种子休眠的因素,破除休眠,寻找其种子快速萌发的方法,以干燥的粗茎秦艽种子为材料,测定种子吸水率及粗提物的活性,使用不同浓度的赤霉素（GA₃）、高锰酸钾（KMnO₄）、聚乙二醇（PEG6000）和过氧化氢（H₂O₂）溶液进行浸种处理,比较不同处理条件对粗茎秦艽种子萌发的影响。结果显示,粗茎秦艽种皮对种子吸水没有明显阻碍作用;不同浓度的种子粗提物对白菜、小麦的萌发和生长均表现出抑制作用;不同浓度的粗提物对粗茎秦艽种子自身的萌发也表现出一定的抑制作用,当粗提物浓度达到0.1 g/mL时,抑制作用最显著（P < 0.05）;高锰酸钾处理可提高粗茎秦艽种子的萌发率,浓度为1.5%时效果较显著（P < 0.05）,而过氧化氢处理对粗茎秦艽种子的萌发效果不如前者,此外,用500 mg/L的赤霉素浸种和300 mg/L的聚乙二醇预处理也可显著打破粗茎秦艽种子休眠（P < 0.01）。研究结果表明粗茎秦艽种子的内源抑制物是影响其休眠的因素之一;种皮的机械阻碍也在一定程度上影响了种子萌发;粗茎秦艽种子具有综合性休眠特性。高锰酸钾预处理、赤霉素浸种和聚乙二醇引发均可打破种子休眠、缩短种子出芽时间,提高种子的发芽能力。     

Abscisic acid inhibits germination of mature Arabidopsis seeds by limiting the availability of energy and nutrients

The addition of abscisic acid (ABA) to mature non-dormant seeds inhibits their germination. This effect of ABA might be related to its natural function as an endogenous inhibitor of precocious germination during seed formation. In this work, we studied how ABA affects the germination of mature seeds and the growth of nascent seedlings of Arabidopsisthaliana (L.) Heynh. Our findings were as follows: (i) inhibition by ABA was gradual, dose-dependent, and did not disappear after germination; (ii) inhibition of germination was relieved by the addition of metabolizable sugars or amino acids to the plating media; (iii) the effect of sugars and amino acids was cooperative, indicating that these two groups of metabolites relieve different deficiencies; (iv) ABA caused appreciable alterations in energy and nitrogen metabolism; and (v) ABA prevented the degradation of the seed storage proteins. In summary, ABA appears to inhibit seed germination by restricting the availability of energy and metabolites. This mechanism seems consistent with other known effects of ABA. Received: 3 February 1997 / Accepted: 10 March 1997     

Requirement for the action of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus

The role of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus L. was investigated. The seeds readily germinated in water and darkness at 24°C. Application of ethylene or of its precursor I-aminocyclopropane-I-carboxylic acid (ACC) slightly increased the rate of germination. Both compounds effectively antagonized osmotic inhibition by polyethyleneglycol. Application of aminoethoxyvinylglycine (AVG) reduced ethylene production by 90% but did not inhibit germination. However, germination was inhibited by 2,5-norbornadiene, a competitive inhibitor of ethylene action. This inhibition was counteracted by ethylene, ethephon or ACC and enforced by AVG. It is concluded that the action of endogenous ethylene is an indispensable factor during germination of non-dormant seeds of A. caudatus. Ethylene action is required from the start of imbibition on. In water, low levels of endogenous ethylene are sufficient for this action. PEG increased the ethylene requirement considerably.     

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.     

Ascorbic acid and reactive oxygen species are involved in the inhibition of seed germination by abscisic acid in rice seeds

The antagonism between abscisic acid (ABA) and gibberellin (GA) plays a key role in controlling seed germination, but the mechanism of antagonism during this process is not known. The possible links among ABA, reactive oxygen species (ROS), ascorbic acid (ASC), and GA during rice seed germination were investigated. Unlike in non-seed tissues where ROS production is increased by ABA, ABA reduced ROS production in imbibed rice seeds, especially in the embryo region. Such reduced ROS also led to an inhibition of ASC production. GA accumulation was also suppressed by a reduced ROS and ASC level, which was indicated by the inhibited expression of GA biosynthesis genes, amylase genes, and enzyme activity. Application of exogenous ASC can partially rescue seed germination from ABA treatment. Production of ASC, which acts as a substrate in GA biosynthesis, was significantly inhibited by lycorine which thus suppressed the accumulation of GA. Consequently, expression of GA biosynthesis genes was suppressed by the low levels of ROS and ASC in ABA-treated seeds. It can be concluded that ABA regulates seed germination in multiple dimensions. ROS and ASC are involved in its inhibition of GA biosynthesis.