Effects of Cycloheximide on Abscisic Acid Biosynthesis and Stomatal Aperture in Bean Leaves

Li, Yongguang; Walton, Daniel C.

doi:10.1104/pp.93.1.128

Cited by 11 publications

(6 citation statements)

References 13 publications

(9 reference statements)

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“…Their effect on the decrease in ABA at 300 min was relatively slight. Cycloheximide also had a small effect at 0.01 mM when infiltrated into a leaf (Table 4), and, as expected from the findings of Guerrero and Mullet (1986) and Li and Walton (1990), it prevented the formation of any stress ABA because it blocked the synthesis of the dioxygenase.…”

Section: Infiltration Of Leaves With Toxic and Osmotic Agentssupporting

confidence: 71%

“…Cycloheximide (1 m M ) also caused a considerable release of free ABA (1.1 µg ABA g −1 leaf) but the mechanism is unclear. It does block protein synthesis by cytosolic ribosomes and Guerrero and Mullet (1986) and Li and Walton (1990) found that it prevented the increase in stress ABA caused by wilting. The large amounts of free ABA, therefore, establish that the biosynthetic, enzymic processes that normally produce the large rise in stress ABA after 2–3 h are not responsible for the production of the ABA released.…”

Section: Resultsmentioning

confidence: 99%

“…Compounds tested on leaf homogenates to find if they caused the release of free ABA included cycloheximide (Table 1) which Guerrero and Mullet (1986) and Li and Walton (1990) had found to block the biosynthesis of stress ABA and thereby established that at least one enzyme of ABA biosynthesis was formed by cytosolic ribosomes. The rapid release of ABA, therefore, is independent of cytoplasmic protein synthesis.…”

Section: Resultsmentioning

confidence: 99%

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Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis

Georgopoulou

Milborrow

2012

Physiologia Plantarum

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The 'fettered' fraction of abscisic acid (ABA) that is held within the chloroplasts of unwilted bean and Commelina communis leaves is released when the leaves wilt and it is this 'free' ABA that is now proposed to cause the stomata to close within 2 or 3 min, well before the rise in total ABA can be detected. The large increase in 'stress' ABA begins 2-3 h later. The fettered ABA in a centrifuged homogenate is released by hyperosmotic solutions of mannitol (0.8 M) and NaCl (0.4 M). Dilute solution of halothane (10 mM) and colchicine (1 mM), the detergent sodium dodecyl sulphate (1 mM) the herbicide 2,4-d (0.1 mM) and dithiothreitol (0.01 mM) also caused ABA to be released. Zeatin (0.01 mM), cumene hydroperoxide (0.01 mM) and CaCl(2) (1 mM) had negligible effects. It was postulated that the ABA released from the chloroplasts by wilting could be the signal that initiates the synthesis of the dioxygenase and other enzymes necessary to produce the rise (up to 40-fold) in the amount of stress ABA that is seen 2-3 h later. To test this hypothesis, a solution of (+)-[(2) H(6) ]ABA was vacuum infiltrated into unwilted Commelina leaves to mimic the rise in ABA caused by wilting and gas chromatography/mass spectrometry of the ABA in the extract after 3 h showed that concentrations of (+)-[(2) H(6) ]ABA of up to 0.3 µM stimulated synthesis of endogenous [(1) H]ABA by 15-fold in the unwilted leaves. A 0.5 µM solution blocked the increase in the amount of ABA formed and also reduced the amount of ABA formed in response to a 0.8 M mannitol solution.

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Section: Infiltration Of Leaves With Toxic and Osmotic Agentssupporting

confidence: 71%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis

Georgopoulou

Milborrow

2012

Physiologia Plantarum

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“…Passive closure at a high degree of dehydration was also observed at 200C in detached leaves previously fed 0.1 mm CH for 2 h and then allowed to dehydrate. CH is known to block water stress-induced ABA accumulation and to inhibit active stomatal closure in bean leaves (12). As reported in Table II, CH-fed leaves, which did not accumulate ABA, had low LDR (2-3 s-cm-') despite a severe water deficit, and stomatal closure occurred only at a very low RWC (53%).…”

Section: Effect Of Exogenously Applied Aba On Chilling Resistance Of mentioning

confidence: 90%

Involvement of Abscisic Acid in Regulating Water Status in Phaseolus vulgaris L. during Chilling

Pardossi¹,

Vernieri²,

Tognoni³

1992

Plant Physiol.

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During the first hours of chilling, bean (Phaseolus vulgaris L., cv Mondragone) seedlings suffer severe water stress and wilt without any significant increase in leaf abscisic acid (ABA) content (P. Aust J Plant Physiol 18: 25-35). Plants regain turgor after 30 to 40 h. We hypothesized that inability to rapidly synthesize ABA at low temperatures contributes to chilling-induced water stress and that turgor recovery after 30 to 40 h is mediated by changes in endogenous ABA content. Entire bean seedlings were subjected to long-term (up to 6 d) chilling (3°C, 0.2-0.4 kPa vapor pressure deficit, 100 Mmol.m-2.s-1 photosynthetic photon flux density, continuous fluorescent light). During the first 24 h, stomata remained open, and plants rapidly wilted as leaf transpiration exceeded root water absorption. During this phase, ABA did not accumulate in leaves or in roots. After 24 h, ABA content increased in both tissues, leaf diffusion resistance increased, and plants rehydrated and regained turgor. No osmotic adjustment was associated with turgor recovery. Following turgor recovery, stomata remained closed, and ABA levels in both roots and leaves were elevated compared with controls. The application of ABA (0.1 mM) to the root system of the plants throughout exposure to 3°C prevented the chilling-induced water stress. Excised leaves fed 0.1 mm ABA via the transpiration stream had greater leaf diffusion resistance at 20 and 3°C compared with non-ABA fed controls, but the amount of ABA needed to elicit a given degree of stomatal closure was higher at 3°C compared with 20°C. These findings suggest that endogenous ABA may play a role in ameliorating plant water status during chilling.

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“…As an inhibitor of stress-induced ABA accumulation we used cycloheximide (CHI) as described by Quarrie and Lister (1984) and Li and Walton (1990).…”

mentioning

confidence: 99%

Drought‐ and ABA‐induced changes in photosynthesis of barley plants

Попова

Tsonev

Lazova

et al. 1996

Physiologia Plantarum

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The changes caused by drought stress and abscisic acid (ABA) on photosynthesis of barley plants (Hordeum vulgare. L. cv. Alfa) have been studied. Drought stress was induced by allowing the leaves to lose 12% of their fresh weight. Cycloheximide (CHI), an inhibitor of stress‐induced ABA accumulation, was used to distinguish alterations in photosynthetic reactions that are induced after drought stress in response to elevated ABA levels from those that are caused directly by altered water relations. Four hoars after imposition of drought stress or 2 h after application of ABA, Ihe bulk of the leaf's ABA content measured by enzyme‐amplified ELISA, increased 14‐ and 16‐fold, respectively. CHI fully blocked the stress‐induced ABA accumulation. Gas exchange measurements and analysis of enzyme activities were used to study the reactions of photosynthesis to drought stress and ABA. Leaf dehydration or ABA treatment led to a noticeable decrease in both the initial slope of the curves representing net photosynthetic rate versus intercellular CO2 concentration and the maximal rate of photosynthesis; dehydration of CHI‐treated plants showed much slower inhibition of the latter. The calculated values of the intercellular CO2 concentration, CO2 compensation point and maximal carboxylating efficiency of ribulose 1,5‐bisphosphate (RuBP) carboxylase support the suggestion that biochemical factors are involved in the response of photosynthesis to ABA and drought stress. RuBP carboxylase activity was almost unaffected in ABA‐ and CHI‐treated, non‐stressed plants. A drop in enzyme activity was observed after leaf dehydration of the control and ABA‐treated plants. When barley plants were supplied with ABA, the activity of carbonic anhydrase (CA, EC 4.2.2.1) increased more than 2‐fold. Subsequent dehydration caused an over 1.5‐fold increase in CA activity of the control plants and a more than 2.5‐fold increase in ABA‐treated plants. Dehydration of CHI‐treated plants caused no change in enzyme activity. It is suggested that increased activity of CA is a photosynthetic response to elevated ABA concentration.

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Effects of Cycloheximide on Abscisic Acid Biosynthesis and Stomatal Aperture in Bean Leaves

Cited by 11 publications

References 13 publications

Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis

Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis

Involvement of Abscisic Acid in Regulating Water Status in Phaseolus vulgaris L. during Chilling

Drought‐ and ABA‐induced changes in photosynthesis of barley plants

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Effects of Cycloheximide on Abscisic Acid Biosynthesis and Stomatal Aperture in Bean Leaves

Cited by 11 publications

References 13 publications

Initiation of the synthesis of ‘stress’ ABA by (+)‐[2H6]ABA infiltrated into leaves of Commelina communis

Initiation of the synthesis of ‘stress’ ABA by (+)‐[2H6]ABA infiltrated into leaves of Commelina communis

Involvement of Abscisic Acid in Regulating Water Status in Phaseolus vulgaris L. during Chilling

Drought‐ and ABA‐induced changes in photosynthesis of barley plants

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Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis

Initiation of the synthesis of ‘stress’ ABA by (+)‐[²H₆]ABA infiltrated into leaves of Commelina communis