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1.
Generation of active pools of abscisic acid revealed by in vivo imaging of water-stressed Arabidopsis 总被引:2,自引:0,他引:2
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A noninvasive, cell-autonomous reporter system was developed to monitor the generation and distribution of physiologically active pools of abscisic acid (ABA). ABA response (abi1-1) and biosynthesis (aba2-1) mutants of Arabidopsis (Arabidopsis thaliana) were used to validate the system in the presence and absence of water stress. In the absence of water stress, low levels of ABA-dependent reporter activation were observed in the columella cells and quiescent center of the root as well as in the vascular tissues and stomata of cotyledons, suggesting a nonstress-related role for ABA in these cell types. Exposure of seedlings to exogenous ABA resulted in a uniform pattern of reporter expression. In marked contrast, reporter expression in response to drought stress was predominantly confined to the vasculature and stomata. Surprisingly, water stress applied to the root system resulted in the generation of ABA pools in the shoot but not in the root. The analysis of the response dynamics revealed a spread of physiologically active ABA from the vascular tissue into the areoles of the cotyledons. Later, ABA preferentially activated gene expression in guard cells. The primary sites of ABA action identified by in planta imaging corresponded to the sites of ABA biosynthesis, i.e. guard cells and cells associated with vascular veins. Hence, water stress recognized by the root system predominantly results in shoot-localized ABA action that culminates in a focused response in guard cells. 相似文献
2.
Tissue-specific localization of an abscisic acid biosynthetic enzyme, AAO3, in Arabidopsis 总被引:6,自引:3,他引:6
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Koiwai H Nakaminami K Seo M Mitsuhashi W Toyomasu T Koshiba T 《Plant physiology》2004,134(4):1697-1707
Arabidopsis aldehyde oxidase 3 (AAO3) is an enzyme involved in abscisic acid (ABA) biosynthesis in response to drought stress. Since the enzyme catalyzes the last step of the pathway, ABA production sites may be determined by the presence of AAO3. Here, AAO3 localization was investigated using AAO3 promoter:AAO3-GFP transgenic plants and by an immunohistochemical technique. AAO3-GFP protein exhibited an activity to produce ABA from abscisic aldehyde, and the transgene restored the wilty phenotype of the aao3 mutant. GFP-fluorescence was detected in the root tips, vascular bundles of roots, hypocotyls and inflorescence stems, and along the leaf veins. Intense immunofluorescence signals were localized in phloem companion cells and xylem parenchyma cells. Faint but significant GFP- and immuno-fluorescence signals were observed in the leaf guard cells. In situ hybridization with antisense AAO3 mRNA showed AAO3 mRNA expression in the guard cells of dehydrated leaves. These results indicate that the ABA synthesized in vascular systems is transported to various target tissues and cells, and also that the guard cells themselves are able to synthesize ABA. 相似文献
3.
The site of action of abscisic acid at the guard cell plasmalemma of Valerianella locusta 总被引:12,自引:5,他引:7
WOLFRAM HARTUNG 《Plant, cell & environment》1983,6(5):427-428
Abstract. Abscisic acid (ABA) is taken up by guard cells of isolated epidermata of Valerianella locusta only at low external pH values. At pH 8.0, when nearly all ABA molecules are present as the union of ABA (ABA− ), no uptake can be observed. ABA-dependent movement of stomata was tested at external pH values between 5.0 and 8.0. Independent of the external pH, ABA induced stomatal closure at all tested ABA concentrations. It is concluded that ABA need not be taken up into the cytosol of the guard cells in order to induce slomatal closure. The primary site of ABA action at the guard cell plasmalemma must be located either at the outer surface of the plasmalemma or at least be easily accessible from outside. ABA− is as effective as undissociated ABA (ABAH). 相似文献
4.
Drought is one of the most severe environmental stresses affecting plant growth and limiting crop production. Although many genes involved in adaptation to drought stress have been disclosed, the relevant molecular mechanisms are far from understood. This study describes an Arabidopsis gene, ASPG1 (ASPARTIC PROTEASE IN GUARD CELL 1), that may function in drought avoidance through abscisic acid (ABA) signalling in guard cells. Overexpression of the ASPG1 gene enhanced ABA sensitivity in guard cells and reduced water loss in ectopically overexpressing ASPG1 (ASPG1-OE) transgenic plants. In ASPG1-OE plants, some downstream targets in ABA and/or drought-signalling pathways were altered at various levels, suggesting the involvement of ASPG1 in ABA-dependent drought avoidance in Arabidopsis. By analysing the activities of several antioxidases including superoxide dismutase and catalase in ASPG1-OE plants, the existence was demonstrated of an effective detoxification system for drought avoidance in these plants. Analysis of ProASPG1-GUS lines showed a predominant guard cell expression pattern in various aerial tissues. Moreover, the protease activity of ASPG1 was characterized in vitro, and two aspartic acid sites, D180 and D379, were found to be key residues for ASPG1 aspartic protease activity in response to ABA. In summary, these findings suggest that functional ASPG1 may be involved in ABA-dependent responsiveness and that overexpression of the ASPG1 gene can confer drought avoidance in Arabidopsis. 相似文献
5.
Desikan R Cheung MK Bright J Henson D Hancock JT Neill SJ 《Journal of experimental botany》2004,55(395):205-212
Increased synthesis and redistribution of the phytohormone abscisic acid (ABA) in response to water deficit stress initiates an intricate network of signalling pathways in guard cells leading to stomatal closure. Despite the large number of ABA signalling intermediates that are known in guard cells, new discoveries are still being made. Recently, the reactive oxygen species hydrogen peroxide (H2O2) and the reactive nitrogen species nitric oxide (NO) have been identified as key molecules regulating ABA-induced stomatal closure in various species. As with many other physiological responses in which H2O2 and NO are involved, stomatal closure in response to ABA also appears to require the tandem synthesis and action of both these signalling molecules. Recent pharmacological and genetic data have identified NADPH oxidase as a source of H2O2, whilst nitrate reductase has been identified as a source of NO in Arabidopsis guard cells. Some signalling components positioned downstream of H2O2 and NO are calcium, protein kinases and cyclic GMP. However, the exact interaction between the various signalling components in response to H2O2 and NO in guard cells remains to be established. 相似文献
6.
ABA-regulated promoter activity in stomatal guard cells 总被引:4,自引:0,他引:4
Jane E. Taylor Kirstie F. Renwick Alex A.R. Webb Martin R. McAinsh Antonella Furini Dorothea Bartels Ralph S. Quatrano William R. Marcotte Jr. Alistair M. Hetherington 《The Plant journal : for cell and molecular biology》1995,7(1):129-134
CDeT6-19 is an ABA-regulated gene which has been isolated from Craterostigma plantagineum . The CDeT6-19 gene promoter has been fused to the β- glucuronidase reporter gene ( GUS ) and used to stably transform Arabidopsis thaliana and Nicotiana tabacum . This construct has been shown to be expressed in stomatal guard cells and often in the adjacent epidermal cells of both species in response to both exogenous ABA and drought stress. These results indicate that the stomatal guard cell is competent to relay an ABA signal to the nucleus. In contrast GUS expression directed by the promoter from a predominantly seed-specific, ABA-regulated gene, Em , or the promoter from the ABA-regulated CDeT27-45 gene is not detectable in the epidermal or guard cells of tobacco or Arabidopsis in response to ABA. The fact that not all ABA-regulated gene promoters are active in stomatal guard cells suggests that effective transduction of the signal is dependent upon particular regions within the gene promoter or that guard cells lack all or part of the specific transduction apparatus required to couple the ABA signal to these promoters. This suggests that there are multiple ABA stimulus response coupling pathways. The identification of a regulatory sequence from an ABA-induced gene which is expressed in stomatal guard cells creates the possibility of examining the role of Ca2+ and other second messengers in ABA-induced gene expression. 相似文献
7.
PHO1 expression in guard cells mediates the stomatal response to abscisic acid in Arabidopsis 总被引:1,自引:0,他引:1
Céline Zimmerli Alain Vavasseur Hubert Bauer Rainer Hedrich Yves Poirier 《The Plant journal : for cell and molecular biology》2012,72(2):199-211
Stomatal opening and closing are driven by ion fluxes that cause changes in guard cell turgor and volume. This process is, in turn, regulated by environmental and hormonal signals, including light and the phytohormone abscisic acid (ABA). Here, we present genetic evidence that expression of PHO1 in guard cells of Arabidopsis thaliana is required for full stomatal responses to ABA. PHO1 is involved in the export of phosphate into the root xylem vessels and, as a result, the pho1 mutant is characterized by low shoot phosphate levels. In leaves, PHO1 was found expressed in guard cells and up‐regulated following treatment with ABA. The pho1 mutant was unaffected in production of reactive oxygen species following ABA treatment, and in stomatal movements in response to light cues, high extracellular calcium, auxin, and fusicoccin. However, stomatal movements in response to ABA treatment were severely impaired, both in terms of induction of closure and inhibition of opening. Micro‐grafting a pho1 shoot scion onto wild‐type rootstock resulted in plants with normal shoot growth and phosphate content, but failed to restore normal stomatal response to ABA treatment. PHO1 knockdown using RNA interference specifically in guard cells of wild‐type plants caused a reduced stomatal response to ABA. In agreement, specific expression of PHO1 in guard cells of pho1 plants complemented the mutant guard cell phenotype and re‐established ABA sensitivity, although full functional complementation was dependent on shoot phosphate sufficiency. Together, these data reveal an important role for phosphate and the action of PHO1 in the stomatal response to ABA. 相似文献
8.
The coronatine-insensitive 1 mutation reveals the hormonal signaling interaction between abscisic acid and methyl jasmonate in Arabidopsis guard cells. Specific impairment of ion channel activation and second messenger production 总被引:3,自引:0,他引:3
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Munemasa S Oda K Watanabe-Sugimoto M Nakamura Y Shimoishi Y Murata Y 《Plant physiology》2007,143(3):1398-1407
Methyl jasmonate (MeJA) elicits stomatal closing similar to abscisic acid (ABA), but whether the two compounds use similar or different signaling mechanisms in guard cells remains to be clarified. We investigated the effects of MeJA and ABA on second messenger production and ion channel activation in guard cells of wild-type Arabidopsis (Arabidopsis thaliana) and MeJA-insensitive coronatine-insensitive 1 (coi1) mutants. The coi1 mutation impaired MeJA-induced stomatal closing but not ABA-induced stomatal closing. MeJA as well as ABA induced production of reactive oxygen species (ROS) and nitric oxide (NO) in wild-type guard cells, whereas MeJA did not induce production of ROS and NO in coi1 guard cells. The experiments using an inhibitor and scavengers demonstrated that both ROS and NO are involved in MeJA-induced stomatal closing as well as ABA-induced stomatal closing. Not only ABA but also MeJA activated slow anion channels and Ca(2+) permeable cation channels in the plasma membrane of wild-type guard cell protoplasts. However, in coi1 guard cell protoplasts, MeJA did not elicit either slow anion currents or Ca(2+) permeable cation currents, but ABA activated both types of ion channels. Furthermore, to elucidate signaling interaction between ABA and MeJA in guard cells, we examined MeJA signaling in ABA-insensitive mutant ABA-insensitive 2 (abi2-1), whose ABA signal transduction cascade has some disruption downstream of ROS production and NO production. MeJA also did not induce stomatal closing but stimulated production of ROS and NO in abi2-1. These results suggest that MeJA triggers stomatal closing via a receptor distinct from the ABA receptor and that the coi1 mutation disrupts MeJA signaling upstream of the blanch point of ABA signaling and MeJA signaling in Arabidopsis guard cells. 相似文献
9.
Heterotrimeric G-protein regulation of ROS signalling and calcium currents in Arabidopsis guard cells 总被引:1,自引:0,他引:1
Heterotrimeric G proteins composed of Gα, Gβ, and Gγ subunits are important signalling agents in both animals and plants. In plants, G proteins modulate numerous responses, including abscisic acid (ABA) and pathogen-associated molecular pattern (PAMP) regulation of guard cell ion channels and stomatal apertures. Previous analyses of mutants deficient in the sole canonical Arabidopsis Gα subunit, GPA1, have shown that Gα-deficient guard cells are impaired in ABA inhibition of K(+) influx channels, and in pH-independent activation of anion efflux channels. ABA-induced Ca(2+) uptake through ROS-activated Ca(2+)-permeable channels in the plasma membrane is another key component of ABA signal transduction in guard cells, but the question of whether these channels are also dependent on Gα for their ABA response has not been evaluated previously. We used two independent Arabidopsis T-DNA null mutant lines, gpa1-3 and gpa1-4, to investigate this issue. We observed that gpa1 mutants are disrupted both in ABA-induced Ca(2+)-channel activation, and in production of reactive oxygen species (ROS) in response to ABA. However, in response to exogenous H(2)O(2) application, I(Ca) channels are activated normally in gpa1 guard cells. In addition, H(2)O(2) inhibition of stomatal opening and promotion of stomatal closure are not disrupted in gpa1 mutant guard cells. These data indicate that absence of GPA1 interrupts ABA signalling between ABA reception and ROS production, with a consequent impairment in Ca(2+)-channel activation. 相似文献
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12.
Phospholipase C is required for the control of stomatal aperture by ABA 总被引:20,自引:0,他引:20
Hunt L Mills LN Pical C Leckie CP Aitken FL Kopka J Mueller-Roeber B McAinsh MR Hetherington AM Gray JE 《The Plant journal : for cell and molecular biology》2003,34(1):47-55
The calcium-releasing second messenger inositol 1,4,5-trisphosphate is involved in the regulation of stomatal aperture by ABA. In other signalling pathways, inositol 1,4,5-trisphosphate is generated by the action of phospholipase C. We have studied the importance of phospholipase C in guard cell ABA-signalling pathways. Immunolocalisation of a calcium-activated phospholipase C confirmed the presence of phospholipase C in tobacco guard cells. Transgenic tobacco plants with considerably reduced levels of phospholipase C in their guard cells were only partially able to regulate their stomatal apertures in response to ABA. These results suggest that phospholipase C is involved in the amplification of the calcium signal responsible for reductions in stomatal aperture in response to ABA. As full ABA-induced inhibition of stomatal opening was not observed, our results support a role for the action of other calcium-releasing second messengers in the guard cell ABA-signalling pathway. It is not known whether these different calcium-releasing second messengers act in the same or parallel ABA-signalling pathways. 相似文献
13.
Evidence for an Extracellular Reception Site for Abscisic Acid in Commelina Guard Cells 总被引:25,自引:7,他引:18
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The phytohormone abscisic acid (ABA) triggers stomatal closing as a physiological response to drought stress. Several basic questions limit an understanding of the mechanism of ABA reception in guard cells. Whether primary ABA receptors are located on the extracellular side of the plasma membrane, within the intracellular space of guard cells, or both remains unknown. Furthermore, it is not clear whether ABA must be transported into guard cells to exert control over stomatal movements. In the present study, a combination of microinjection into guard cells and physiological assays of stomatal movements have been performed to determine primary sites of ABA reception in guard cells. Microinjection of ABA into guard cells of Commelina communis L. resulted in injected cytosolic concentrations of 50 to 200 [mu]M ABA and in additional experiments in lower concentrations of approximately 1 [mu]M ABA. Stomata with ABA-loaded guard cells (n > 180) showed opening similar to stomata with uninjected guard cells. The viability of guard cells following ABA injection was demonstrated by neutral red staining as well as monitoring of stomatal opening. Extracellular application of 10 [mu]M ABA inhibited stomatal opening by 98% at pH 6.15 and by 57% at pH 8.0. The pH dependence of extracellular ABA action may suggest a contribution of an intracellular ABA receptor to stomatal regulation. The findings presented here show that intracellular ABA alone does not suffice to inhibit stomatal opening under the imposed conditions. Furthermore, these data provide evidence that a reception site for ABA-mediated inhibition of stomatal opening is on the extracellular side of the plasma membrane of guard cells. 相似文献
14.
15.
Abscisic acid (ABA) is a phytohormone that responds to environmental stresses, such as water deficiency. Recent studies have shown that ABA biosynthetic enzymes are expressed in the vascular area under both nonstressed and water-stressed growth conditions. However, specific cells in the vasculature involved in ABA biosynthesis have not been identified. Here, we detected the expression of two genes encoding ABA biosynthetic enzymes, ABSCISIC ACID DEFICIENT2 and ABSCISIC ALDEHYDE OXIDASE3, in phloem companion cells in vascular tissues. Furthermore, we identified an ATP-binding cassette transporter, Arabidopsis thaliana ABCG25 (AtABCG25), expressed in the same cells. Additionally, AtABCG25-expressing Spodoptera frugiperda9 culture cells showed an ABA efflux function. Finally, we observed that enhancement of ABA biosynthesis in phloem companion cells induced guard cell responses, even under normal growth conditions. These results show that ABA is synthesized in specific cells and can be transported to target cells in different tissues.Hormones are chemical substances that exert a biochemical action on target cells at low concentrations. All multicellular organisms, including animals and plants, produce hormones to control their physiological status. In animals, many ordinary hormones are secreted from specific cells (such as endocrine cells) and transported to their target sites in other areas of the body. However, it remains unclear whether the concept of hormones as defined in animals is applicable to plants, because plant hormones are not generally synthesized in specific cells but are broadly produced (Weyers and Paterson, 2001; Gaspar et al., 2003; Forestan and Varotto, 2012).Abscisic acid (ABA) is a key phytohormone that prevents water loss from the plant body by acting on guard cells, of which stomata (epidermal pores) compose the aerial organs in plants (Hetherington, 2001; Schroeder et al., 2001; Fan et al., 2004; Joshi-Saha et al., 2011). Gene and protein expression analyses using antisense RNA or antibodies specific for ABA biosynthetic enzymes in Arabidopsis (Arabidopsis thaliana) have shown that parenchyma cells in vascular bundles are the abundant expression site of ABA biosynthesis under drought stress and well-watered growth conditions (Cheng et al., 2002; Koiwai et al., 2004; Endo et al., 2008). Because the vasculature is separated from guard cells, it has been suggested that ABA is transported from the site of synthesis to the site of action (Seo and Koshiba, 2011).We previously found that an ATP-binding cassette (ABC) transporter family, Arabidopsis thaliana ABCG25 (AtABCG25), is expressed mainly in vascular tissues, and it is expected to function as an ABA exporter that transports ABA from inside to outside cells (Kuromori et al., 2010). According to this observation, we have proposed a working model: ABA is exported from ABA-synthesizing cells in vascular tissues by AtABCG25 to reach distant guard cells and induce stomatal closure (Kuromori and Shinozaki, 2010; Umezawa et al., 2010, 2011). However, the parenchyma cells in vascular tissues expressing ABA biosynthetic enzymes or AtABCG25 transporting factor have not been identified.Here, we explored whether specific cells express ABA biosynthetic enzymes or an ABA transporter and found that their genes were expressed in phloem companion cells of vascular tissues. ABA synthesis in these cells enhances transsignaling to distant guard cells of the epidermis. These results show that ABA is synthesized in specific cells and transported to target cells in another tissue. This result is similar to endocrine hormones in animals and suggests that the ABA transport pathway between tissues in plants may be associated with specific transporters. 相似文献
16.
Regulation of the ABA-sensitive Arabidopsis potassium channel gene GORK in response to water stress 总被引:8,自引:0,他引:8
Becker D Hoth S Ache P Wenkel S Roelfsema MR Meyerhoff O Hartung W Hedrich R 《FEBS letters》2003,554(1-2):119-126
17.
Wenxiu Ye Mohammad Anowar Hossain Shintaro Munemasa Yoshimasa Nakamura Izumi C. Mori Yoshiyuki Murata 《Journal of plant physiology》2013
We recently demonstrated that endogenous abscisic acid (ABA) is involved in methyl jasmonate (MeJA)-induced stomatal closure in Arabidopsis thaliana. In this study, we investigated whether endogenous ABA is involved in MeJA-induced reactive oxygen species (ROS) and nitric oxide (NO) production and cytosolic alkalization in guard cells using an ABA-deficient Arabidopsis mutant, aba2-2, and an inhibitor of ABA biosynthesis, fluridon (FLU). The aba2-2 mutation impaired MeJA-induced ROS and NO production. FLU inhibited MeJA-induced ROS production in wild-type guard cells. Pretreatment with 0.1 μM ABA, which does not induce stomatal closure in the wild type, complemented the insensitivity to MeJA of the aba2-2 mutant. However, MeJA induced cytosolic alkalization in both wild-type and aba2-2 guard cells. These results suggest that endogenous ABA is involved in MeJA-induced ROS and NO production but not in MeJA-induced cytosolic alkalization in Arabidopsis guard cells. 相似文献
18.
Arabidopsis OST1 protein kinase mediates the regulation of stomatal aperture by abscisic acid and acts upstream of reactive oxygen species production 总被引:30,自引:0,他引:30
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During drought, the plant hormone abscisic acid (ABA) triggers stomatal closure, thus reducing water loss. Using infrared thermography, we isolated two allelic Arabidopsis mutants (ost1-1 and ost1-2) impaired in the ability to limit their transpiration upon drought. These recessive ost1 mutations disrupted ABA induction of stomatal closure as well as ABA inhibition of light-induced stomatal opening. By contrast, the ost1 mutations did not affect stomatal regulation by light or CO(2), suggesting that OST1 is involved specifically in ABA signaling. The OST1 gene was isolated by positional cloning and was found to be expressed in stomatal guard cells and vascular tissue. In-gel assays indicated that OST1 is an ABA-activated protein kinase related to the Vicia faba ABA-activated protein kinase (AAPK). Reactive oxygen species (ROS) were shown recently to be an essential intermediate in guard cell ABA signaling. ABA-induced ROS production was disrupted in ost1 guard cells, whereas applied H(2)O(2) or calcium elicited the same degree of stomatal closure in ost1 as in the wild type. These results suggest that OST1 acts in the interval between ABA perception and ROS production. The relative positions of ost1 and the other ABA-insensitive mutations in the ABA signaling network (abi1-1, abi2-1, and gca2) are discussed. 相似文献
19.
Arabidopsis mutants of AtABCG22, an ABC transporter gene, increase water transpiration and drought susceptibility 总被引:1,自引:0,他引:1
Kuromori T Sugimoto E Shinozaki K 《The Plant journal : for cell and molecular biology》2011,67(5):885-894
In plants, water vapour is released into the atmosphere through stomata in a process called transpiration. Abscisic acid (ABA) is a key phytohormone that facilitates stomatal closure through its action on guard cells. Recently, ATP-binding cassette (ABC) transporter genes, AtABCG25 and AtABCG40, were shown to be involved in ABA transport and responses. However, the functions of many other AtABCG family genes are still unknown. Here, we identified another ABCG gene (AtABCG22) that is required for stomatal regulation in Arabidopsis. The atabcg22 mutant plants had lower leaf temperatures and increased water loss, implying elevated transpiration through an influence on stomatal regulation. We also found that atabcg22 plants were more suspectible to drought stress than wild-type plants. AtABCG22 was expressed in aerial organs, mainly guard cells, in which the gene expression pattern was consistent with the mutant phenotypes. Using double mutants, we investigated the genetic relationships between the mutations. The atabcg22 mutation further increased the water loss of srk2e/ost1 mutants, which were defective in ABA signalling in guard cells. Also, the atabcg22 mutation enhanced the phenotype of nced3 mutants, which were defective in ABA biosynthesis. Accordingly, the additive roles of AtABCG22 functions in ABA signalling and ABA biosynthesis are discussed. 相似文献
20.
ABA-deficient (aba1) and ABA-insensitive (abi1-1, abi2-1) mutants of Arabidopsis have a wild-type stomatal response to humidity 总被引:3,自引:2,他引:1
In most plant species, a decrease in atmospheric humidity at the leaf surface triggers a decrease in stomatal conductance. While guard cells appear to respond to humidity‐induced changes in transpiration rate, as opposed to relative humidity or vapour pressure difference, the underlying cellular mechanisms for this response remain unknown. In the present set of experiments, abscisic acid (ABA)‐deficient (aba1) and ABA‐insensitive (abi1‐1 and abi2‐1) mutants of Arabidopsis thaliana were used to test the hypothesis that the humidity signal is transduced by changes in the flux or concentration of ABA delivered to the stomatal complex in the transpiration stream. In gas exchange experiments, stomatal conductance was as sensitive to changes in vapour pressure difference in aba1, abi1‐1 and abi2‐1 mutant plants as in wild‐type plants. These experiments appear to rule out an obligate role for either the concentration or flux of ABA or ABA conjugates as mediators of the guard cell response to atmospheric water potential. The results stand in contrast to the well‐established role of ABA in mediating guard cell responses to decreases in soil water potential. 相似文献