Carotenoids and abscisic acid (ABA) biosynthesis in higher plants

Recent research has revealed that abscisic acid (ABA), synthesised in response to water stress, is an apo-carotenoid. Two potential carotenoid precursors, 9'- cis -neoxanthin and 9- cis -violaxanthin, have been identified in light-grown and etiolated leaves, and in roots of a variety of species. Experiments utilizing etiolated Phaseolus vulgaris leaves and deuterium oxide strongly suggest that 9'- cis -neoxanthin, synthesised from all- trans -violaxanthin, is the immediate pre-cleavage precursor of ABA. The cleavage of 9'- cis -neoxanthin, performed by an inducible and specific dioxygenase, is likely to be the rate-limiting step in ABA biosynthesis. Any apocarotenoids formed as by-products of cleavage are probably rapidly degraded by lipoxygenase or related enzymes. After cleavage xanthoxin is converted via ABA-aldehyde to ABA by constitutive enzymes in the cytosol.     

Effects of abscisic acid on ozone tolerance of rice (Oryza sativa L.) seedlings

Ozone is one of the major gaseous pollutants detrimental to crop growthand metabolism. The objective of this research was to study how ABA amelioratesthe effects of ozone on rice seedlings. Seedlings of two rice cultivars withdifferent sensitivities to ozone (Tainung 67, tolerant; and Taichung Native 1,sensitive) were treated with 400 ppb of ozone or ABA and 400ppb of ozone to determine their effect on growth, stomatalmovement, chlorophyll characteristics, and the activity of antioxidant enzymes.Activities of the enzymes SOD, APOD, GR and POD were significantly higher inthesensitive cultivar, TN 1, than in the tolerant cultivar, TNG 67. Seedlings ofthe sensitive cultivar pretreated with ABA (10 M) weresignificantly more tolerant of ozone than control seedlings. Pretreatment withABA effectively reduced stomatal conductance and the degree of injury. Abscisicacid also increased ascorbate peroxidase and glutathione reductase activity.Ozone increased peroxidase activity in sensitive seedlings, but ABA decreasedperoxidase activity. The sensitive cultivar had a higher density of stomata onits leaves than the tolerant cultivar. The results suggest that ABA inducedtolerance to ozone may be more associated with its effects on stomatal movementthan on the modulation of antioxidant enzyme activity.     

Proteomic analysis of phosphoproteins regulated by abscisic acid in rice leaves

Abscisic acid (ABA) is a hormone that regulates plant development and adaptation to environmental stresses. Protein phosphorylation has been recognized as an important mechanism for ABA signaling. However, the target phosphoproteins regulated by ABA are still largely unknown. Here, we report the identification of ABA-regulated phosphoproteins in rice using proteomic approaches. Six ABA-regulated phosphoproteins were identified as G protein beta subunit-like protein, ascorbate peroxidase, manganese superoxide dismutase, triosephosphate isomerase, putative Ca²⁺/H⁺ antiporter regulator protein, and glyoxysomal malate dehydrogenase. These results provide new insight into the regulatory mechanism for some ABA signaling proteins and implicate several previously unrecognized proteins in ABA action.     

Long-distance signalling of abscisic acid (ABA): the factors regulating the intensity of the ABA signal

Abscisic acid (ABA) is a stress signal, which moves in the xylem from the roots to the aerial parts of the plant, where it regulates stomatal movement and the activity of shoot meristems. Root growth-promoting microorganisms in the rhizosphere, lateral ABA flows in the root cortex across apoplastic barriers, ABA redistribution in the stem, leaf apoplastic pH values, and the action of beta-glucosidases, both in the apoplast and the cytosol of the mesophyll, play an important role in the regulation of signal intensity. The significance of abscisic acid glucose ester as a long-distance stress signal is discussed.     

Effects of abscisic acid and its related compounds on rice seedling growth

Rice seedlings with the mesocotyl and coleoptile (the undeveloped leaves enclosed in the coleoptile) are here referred to as MC type seedlings and are considered to be suitable for deep sowing. We investigated the effects of abscisic acid (ABA) and several of its related compounds on the occurrence of MC type seedlings and on rice mesocotyl growth. Rice (Oryza sativa L. cv. JC 91) seedlings were grown on 0.8% agar medium in the presence or absence of various kinds of ABAs under aseptic conditions at 30 °C in the dark for 14 days. The activity of the R isomer of ABA (R-ABA) was slightly less than that of the naturally occurring S form (S-ABA) concerning the occurrence of MC type rice seedlings and the growth of the rice mesocotyl. In addition, the racemate of R-and S-ABA (RS-ABA) is less effective than R-ABA and S-ABA alone.Trans-ABA had no activity in relation to both percent occurrence of MC type seedlings and mesocotyl growth. The results of the present study suggest that the occurrence of MC type rice seedlings and the growth of rice mesocotyls were closely related to structure-activity relationships with analogs of ABA.     

Involvement of abscisic acid (ABA) in bulb dormancy of Allium wakegi Araki I. Endogenous levels of ABA in relation to bulb dormancy and effects of exogenous ABA and fluridone

Allium wakegi plants exposed to long days (LD, 14 h-photoperiod) developed bulbs, which were dormant from the 30th to the 125th day of LD, but those grown under natural short days (SD) did not develop bulbs. The contents of abscisic acid (ABA) in both whole bulbs and buds of the bulbs increased in LD, reaching a maximum at the 60th day of LD and decreasing thereafter, but those in basal leaf sheaths (this part corresponds to a bulb after bulb development) and buds did not increase in SD. The ABA content was related to the depth of bulb dormancy. Application of 500 M ABA to bulbs for 24 h significantly delayed sprouting, but that of 5 or 50 M ABA had little or no effect. Application of 25 or 125 M fluridone to the soil just before exposure to LD bleached new expanding leaves and reduced bulb size, but had no effect on the development of bulb scales that characterize bulb formation. The bulbs formed under such conditions sprouted earlier than those of control plants. The levels of endogenous ABA in bulbs, buds of the bulbs, leaf blades, and roots were reduced by fluridone application. These results indicate that ABA plays an important role in bulb dormancy of Allium wakegi.     

Synthesis and biological activity of abscisic acid esters

16 ABA esters including 11 new compounds were prepared by two different esterification routes. All the structures of ABA esters were confirmed by ¹H NMR, ¹³C NMR and HRMS. Their biological activity and hydrolysis stability were investigated. Fortunately, there were 15 and 9 compounds which displayed much better or nearly the same inhibition activity for rice seedling growth and Arabidopsis thaliana seed germination compared to ABA, respectively. Especially, compounds 2d and 2g showed better biological activities than ABA in the three tests. Moreover, we found that chemical hydrolysis ability of the esters in vitro had little relationship to their biological activity.     

Effects of exogenous abscisic acid (ABA) on cucumber seedling leaf carbohydrate metabolism under low temperature

Seedlings with four true leaves of cucumbers (Cucumis sativus L.), Guonong No.25 (a cold-tolerant cultivar) and Guonong No.41 (a cold sensitive cultivar), were grown under normal or low temperature conditions: 25°C/18°C or 15°C/8°C (day/night). The seedlings of Guonong No.25 under low temperature were also treated with or without exogenous ABA. The purpose of our study was to find out the effects of low temperature and exogenous ABA application on the carbohydrate metabolism in the cucumber plants. Time course changes of carbohydrate contents and activities of stachyose synthase and alkaline α-galactosidase in the seedling leaves were investigated after the treatment. Our results show that compared to the seedlings under temperatures of 25°C/18°C, the seedlings of the both tested genotypes under 15°C/8°C (day/night) have significantly higher contents of all measured soluble carbohydrates. Significant difference in stachyose synthase activity is observed between the two genotypes under normal temperature or low temperature. Under normal temperature, leaf stachyose synthase activity in Guonong No.41 is higher than that in Guonong No.25. The stachyose synthase activity of Guonong No.41 decreases sharply under low temperature, but that of Guonong No.25 increases 3 days after treatment and then decreases to the original level. In contrast, there is no significant genotypic difference in alkaline α-galactosidase activity. Additionally, compared to the control seedlings treated with 0 μM ABA, the seedlings treated with 50 and 150 μM ABA accumulate substantial amounts of all tested soluble carbohydrates except galactose whereas 250 μM ABA treated seedlings show decreased levels of all these soluble carbohydrates. Stachyose synthase activity increases significantly upon 50 and 150 μM ABA treatments. Fan-zhen Menga, Li-ping Hu, and Shao-hui Wang contributed equally to the paper.     

Antagonistic effects of abscisic acid and gibberellic acid on the breaking of dormancy of Fagus sylvatica seeds

Study of the factors involved in the dormancy of Fagus sylvatica seeds shows that such dormancy is due partly to the seed coats and partly to endogenous factors. Seed coat removal accelerates both the release from dormancy and the effects of the other treatments that abolish it. The dormancy of these seeds is eliminated by cold treatment at 4°C over a period longer than 8 weeks, and exogenous application of abscisic acid (ABA) reverses the effects of low temperature, the seeds remaining in an ungerminated state. Additionally, ABA reduces protein synthesis but slightly increases RNA synthesis, which suggests its involvement in the synthesis of RNAs related to this process. In vitro translation of the RNAs isolated from these seeds shows that ABA delays the disappearance of at least 2 polypeptides (of ca 22 and 24 kDa), which are abundant in dormant seeds and under conditions that prevent the release from dormancy, but which disappear under treatments that abolish it. Exogenous application of gibberellic acid (GA₃) proved to be efficient in breaking the dormancy of these seeds and in substituting for cold treatment as well as in antagonizing the effects of ABA on the synthesis of both DNA and proteins. GA₃ also accelerates the disappearance of the two polypeptides abundant in dormant seeds and in ABA-treated seeds. These findings suggest that both ABA and GA₃ could be involved in the regulation of nucleic acid and protein metabolism during dormancy, acting antagonistically in these processes and, specifically, in the regulation of the synthesis of the two proteins that appear to play a role in the maintenance of dormancy in these seeds.     

The effects of gibberellic acid,fluridone, abscisic acid and paclobutrazol on anther culture of Brussels sprouts

Both gibberellic acid (GA₃) and fluridone, a non-specific inhibitor of ABA biosynthesis, promoted embryo production in anther cultures of Brussels sprouts cv. Hal, but not in cv. Gower. Abscisic acid (ABA) and the gibberellin-biosynthesis inhibitor paclobutrazol inhibited embryo production in both cultivars.     

Analysis of an abscisic acid (ABA)-responsive gene promoter belonging to the Asr gene family from tomato in homologous and heterologous systems

Asr is a family of genes that maps to chromosome 4 of tomato. Asr2, a recently reported member of this family, is believed to be regulated by abscisic acid (ABA), stress and ripening. A genomic Asr2 clone has been fully sequenced, and candidate upstream regulatory elements have been identified. To prove that the promoter region is functional in vivo, we fused it upstream of the β-glucuronidase (GUS) reporter gene. The resulting chimeric gene fusion was used for transient expression assays in papaya embryogenic calli and leaves. In addition, the same construct was used to produce transgenic tomato, papaya, tobacco, and potato plants. Asr2 upstream sequences showed promoter function in all of these systems. Under the experimental conditions tested, ABA stimulated GUS expression in papaya and tobacco, but not in tomato and potato systems. Received: 24 March 1997 / Accepted: 26 November 1997     

The effects of two abscisic acid analogues, WL19224 and WL19377, on stomatal closure

The effect on stomatal closure by ABA and its analogues, WL19224 and WL19377 was investigated. The rate of closure showed a sigmoid curve when various concentrations of ABA were applied. A concentration of 10^-9 M ABA was the threshold for stomatal closure; maximal closure occurred at higher concentrations (from 10^-6 M to 10^-3 M). Use of the analogue WL19224 resulted in similar closure responses. However, ABA was more effective at lower concentrations. For example, at 10^-3 M of either WL19224 and ABA, stomata closed to 2.2 μm and about 3 μm, respectively. In contrast, applications of the ABA analogue WL19377 had no effect on stomatal closure. In fact, at concentrations of WL19377 higher than 10^-4 M, stomata were stimulated to open, to about 10% of their initial size. Likewise, applications of WL19377 along with ABA, inhibited ABA-induced stomatal closure. This inhibition was linearly related to the concentrations of the compounds applied. In conclusion, the structural requirements for biological activity of ABA and its analogues cannot be considered individually, but must be assessed for their roles as part of an entire functional group. Although compounds may have similar structures, their ability to control certain physiological activities may be quite different.     

Individual and interactive effects of temperature,carbon dioxide and abscisic acid on mung bean (Vigna radiata) plants

We studied the effects of temperature, carbon dioxide and abscisic acid on mung bean (Vigna radiata). Plants were grown under 26/22°C or 32/28°C (16?h?light/8?h?dark) at 400 or 700?μmol?mol^?1 CO₂ and received ABA application of 0 or 100?μl (10?μg) every other day for three weeks, after eight days of initial growth, in growth chambers. We measured 24 parameters. As individual factors, in 16 cases temperature; in 8 cases CO₂; in 9 cases ABA; and as interactive factors, in 4 cases, each of temperature?×?CO₂, and CO₂?×?ABA; and in 2 cases, temperature?×?ABA were significant. Higher temperatures increased growth, aboveground biomass, growth indices, photochemical quenching (qP) and nitrogen balance index (NBI). Elevated CO₂ increased growth and aboveground biomass. ABA decreased growth, belowground biomass, qP and flavonoids; increased shoot/root mass ratio, chlorophyll and NBI; and had little role in regulating temperature–CO₂ effects.

Abbreviations: A_N: net CO₂ assimilation; E: transpiration; F_v/F_m: maximum quantum yield of PSII; g_s: stomatal conductance; LAR: leaf area ratio; LMA: leaf mass per area; LMR: leaf mass ratio;φPSII: effective quantum yield of PSII; qNP: non-photochemical quenching; qP: photochemical quenching; SRMR: shoot to root mass ratio; WUE: water use efficiency     

A comparative study of the effects of abscisic acid and methyl jasmonate on seedling growth of rice

The effects of abscisic acid (ABA) and methyl jasmonate (MJ) on growth of rice seedlings were compared. The lowest tested concentration of ABA and MJ that inhibited seedling growth was found to be 4.5 and 0.9 µM, respectively. Growth inhibition by ABA is reversible, whereas that by MJ is irreversible. GA₃ was found to be more effective in reversing inhibition of shoot growth by ABA than by MJ. KCl partially relieved MJ-inhibited, but not ABA-inhibited, growth of rice seedlings. The beneficial effect of K⁺ on growth of rice seedlings in MJ medium could not be replaced by Li⁺, Na⁺ or Cs⁺. MJ treatment caused a marked release of K⁺ into the medium. In order to understand whether cell wall-bound peroxidase activity was inversely related to rice seedling growth, effects of ABA and MJ on cell wall-bound peroxidase activity were also examined. Results indicated that both ABA and MJ increased cell wall-bound peroxidase activity in roots and shoots of rice seedlings. Although MJ (4.5 µM) was less effective in inhibiting root growth than ABA (9 µM), MJ was found to increase more cell wall-bound peroxidase activity in roots than ABA.     

Inducible effects of abscisic acid on sporophyte discs from Laminaria japonica Areschoug (Laminariales, Phaeophyceae)

The effects of exogenous abscisic acid (ABA;10^–7–10^–5 M), a knownplantgrowth regulator, on reproduction and growth were investigated by culturingdiscs from sporophytes of Laminaria japonica Areschoug.ABAplays a role in triggering sorus formation, and it was found that sorusformation of discs was fastest in 10^–5 M ABA. Theapplication of 10^–5 M ABA to culturing discs alsosuppressed the expansion of surface area. ABA contents in sorus and vegetativeparts of the sporophyte were determined by bioassay. The mean ABA content insorus parts obtained from sporophytes was 0.222 ± 0.053g equivalent-ABA g wet weight^–1, which wasabout five times higher than the content found in vegetative parts (0.048± 0.009 g equivalent-ABA g wetweight^–1). Taken together these results suggest that sorusdevelopment requires an elevated level of ABA and is associated with decreasingvegetative growth and that the ABA level of the sporophyte may play a crucialrole in reproduction.     

Interactions between ethylene, gibberellin and abscisic acid regulate emergence and growth rate of adventitious roots in deepwater rice

Growth of adventitious roots is induced in deepwater rice (Oryza sativa L.) when plants become submerged. Ethylene which accumulates in flooded plant parts is responsible for root growth induction. Gibberellin (GA) is ineffective on its own but acts in a synergistic manner together with ethylene to promote the number of penetrating roots and the growth rate of emerged roots. Studies with the GA biosynthesis inhibitor paclobutrazol revealed that root emergence was dependent on GA activity. Abscisic acid (ABA) acted as a competitive inhibitor of GA activity. Root growth rate on the other hand was dependent on GA concentration and ABA acted as a potent inhibitor possibly of GA but also of ethylene signaling. The results indicated that root emergence and elongation are distinct phases of adventitious root growth that are regulated through different networking between ethylene, GA and ABA signaling pathways. Adventitious root emergence must be coordinated with programmed death of epidermal cells which cover root primordia. Epidermal cell death is also controlled by ethylene, GA and ABA albeit with cell-type specific cross-talk. Different interactions between the same hormones may be a means to ensure proper timing of cell death and root emergence and to adjust the growth rate of emerged adventitious roots.     

Chemotaxonomic characterization of rice seedling blight complex using fatty acid methyl ester (FAME) profiles

Rice seedling blight is an important disease caused by a complex of fungi that include Fusarium, Rhizopus, Pythium, and Trichoderma species. A modified MIDI method was used for extraction of fatty acids from these causal pathogens, and fatty acid methyl ester (FAME) profiles were characterized. Factors that might affect fatty acid production, such as period of culture and saponification in extraction, were also evaluated. A total of 14 fatty acids were detected, and FAME profiles showed quantitative and qualitative variations by discriminant analysis and principal component analysis. Genus-specific FAME profiles consisting of the types of fatty acid produced and remarkable components of individual fatty acids were observed. The possibility of application as chemotaxonomic methods based on the FAME profiles for diagnosis of the rice seedling blight complex is also discussed.     

Abscisic acid pretreatment enhances salt tolerance of rice seedlings: Proteomic evidence

Enhanced salt tolerance of rice seedlings by abscisic acid (ABA) pretreatment was observed from phenotypic and physiological analyses. Total proteins from rice roots treated with ABA plus subsequent salt stress were analyzed by using proteomics method. Results showed that, 40 protein spots were uniquely upregulated in the seedlings under the condition of ABA pretreatment plus subsequent salt stress, whereas only 16 under the condition of salt treatment. About 78% (31 spots) of the 40 protein spots were only upregulated in the presence of the subsequent salt stress, indicating that plants might have an economical strategy to prevent energy loss under a false alarm. The results also showed that more enzymes involved in energy metabolism, defense, primary metabolism, etc. were upregulated uniquely in ABA-pretreated rice seedlings, suggesting more abundant energy supply, more active anabolism (nitrogen, nucleotide acid, carbohydrate, etc), and more comprehensive defense systems in ABA-pretreated seedlings than in salt stressed ones.     

Effect of zeatin on the growth and indolyl-3-acetic acid and abscisic acid levels in maize roots

Elongation, indolyl-3-acetic acid (IAA) and abscisic acid (ABA) levels, – gas chromatography-mass spectrometry quantification –, in the elongating zone were analysed for maize ( Zea mays L., Cv. LG11) roots immersed in buffer solution with or without zeatin (Z). The effect of Z depends on the initial extension rate of roots. The slower growing roots are more strongly inhibited by Z (10⁻⁷−10₋₅ M ) and they show a greater increase in IAA and ABA content. When compared to the rapidly growing roots, the larger reactivity of the 'slow'ones cannot be attributed to a higher Z uptake as shown when using [¹⁴C]-Z. It is suggested that Z could regulate root elongation by acting on the IAA and/or ABA level. The comparative action of these two hormones is discussed.