Crossref journal-article
American Association for the Advancement of Science (AAAS)
Science (221)
Abstract

The phytohormone abscisic acid (ABA) promotes plant water conservation by decreasing the apertures of stomatal pores in the epidermis through which water loss occurs. We found that Arabidopsis thaliana plants harboring transferred DNA insertional mutations in the sole prototypical heterotrimeric GTP-binding (G) protein α subunit gene, GPA1 , lack both ABA inhibition of guard cell inward K + channels and pH-independent ABA activation of anion channels. Stomatal opening in gpa1 plants is insensitive to inhibition by ABA, and the rate of water loss from gpa1 mutants is greater than that from wild-type plants. Manipulation of G protein status in guard cells may provide a mechanism for controlling plant water balance.

Bibliography

Wang, X.-Q., Ullah, H., Jones, A. M., & Assmann, S. M. (2001). G Protein Regulation of Ion Channels and Abscisic Acid Signaling in Arabidopsis Guard Cells. Science, 292(5524), 2070–2072.

Authors 4
  1. Xi-Qing Wang (first)
  2. Hemayet Ullah (additional)
  3. Alan M. Jones (additional)
  4. Sarah M. Assmann (additional)
References 49 Referenced 410
  1. 10.1146/annurev.ph.52.030190.001213
  2. 10.1152/physrev.1995.75.4.865
  3. 10.1016/0968-0004(92)90005-T
  4. 10.1016/0092-8674(91)90358-6
  5. 10.1007/BF01160403
  6. Lemtri-Chlieh F., MacRobbie E. A. C., J. Membr. Biol. 137, 99 (1994). / J. Membr. Biol. by Lemtri-Chlieh F. (1994)
  7. 10.1104/pp.118.4.1421
  8. 10.1104/pp.110.3.987
  9. 10.1073/pnas.96.21.12192
  10. 10.1073/pnas.96.4.1779
  11. 10.1038/346769a0
  12. 10.1038/343186a0
  13. 10.1073/pnas.95.26.15837
  14. 10.1073/pnas.220417197
  15. 10.1038/338427a0
  16. Allen G. J., Kuchitsu K., Chu S. P., Murata Y., Schroeder J. I., Plant Cell 11, 1785 (1999). / Plant Cell by Allen G. J. (1999)
  17. 10.1073/pnas.87.10.3821
  18. 10.1007/BF00016493
  19. RNA was extracted from guard cell (purity percentage of 99:1 guard cell protoplasts:mesophyll cell protoplasts) and mesophyll cell protoplast preparations. Reverse transcription followed by polymerase chain reaction (PCR) was performed [SuperScript first-strand synthesis system for reverse transcriptase (RT)–PCR Gibco BRL Life Technologies]. The forward primer started at 662 base pairs (bp) in the GPA1 cDNA clone (forward; 5′ ggc tgc tga aat cga aag ac 3′); the reverse primer started at 1266 bp (reverse; 5′ gtc cac cca cgt caa aca at 3′).
  20. 10.1016/S0014-5793(98)00129-X
  21. S. M. Assmann in Signal Transduction in Plant Growth and Development D. P. S. Verma Ed. (Springer Vienna 1996) pp. 39–61. (10.1007/978-3-7091-7474-6_2)
  22. 10.1126/science.1059040
  23. 10.1093/oxfordjournals.pcp.a029314
  24. We used 4- to 6-week-old plants grown under short days. Seeds were germinated in a standard medium (23) in 7.5% (w/v) phytagel (Sigma) with kanamycin (50 mg/liter) for mutants. After 10 days robust seedlings were transferred to Metromix 250 potting mixture (Scotts Marysville OH).
  25. Leymarie J., Lasceve G., Vavasseur A., Aust. J. Plant Physiol. 25, 785 (1998). / Aust. J. Plant Physiol. by Leymarie J. (1998)
  26. Light-induced stomatal opening was assayed according to (25). Normalization of aperture widths by stomatal length (16) mirrored the results shown here for widths alone (48).
  27. Guard cell protoplasts were isolated from rosette leaves of 4- to 5-week-old nonbolting plants [after (7)] except that the second enzyme solution contained 1.3% (w/v) Cellulase RS (Yakult Honsha Tokyo) and 0.0075% (w/v) Pectolyase Y-23 (Seishin Pharmaceutical Tokyo). Solutions used in patch-clamp experiments on K + channels were as follows: 10 mM MES (tris pH 6.0) 10 mM K-glutamate 4 mM MgCl 2 and 1 mM CaCl 2 [osmolality of 540 mmol/kg (bath solution)] and 10 mM Hepes-tris (pH 7.8) 80 mM K-glutamate and 20 mM KCl [osmolality of 560 mmol/kg (pipette solution)]. Fresh adenosine triphosphate (ATP) (5 mM from a 0.5 M Mg ATP stock solution in 0.5 M tris) was added daily to the pipette solution. Final osmolalities were obtained by addition of sorbitol. Time-activated currents were calculated as the difference between average steady-state current between 3000 and 3800 ms and instantaneous current at 20 ms.
  28. 10.1126/science.287.5451.300
  29. Pei Z.-M., Kuchitsu K., Ward J. M., Schwarz M., Schroeder J. I., Plant Cell 9, 409 (1997). / Plant Cell by Pei Z.-M. (1997)
  30. 10.1046/j.1365-313X.1997.12010203.x
  31. The initial pipette solution for anion channel recording was as follows: 150 mM CsCl 2 mM MgCl 2 6.7 mM EGTA 3.35 mM CaCl 2 10 mM Hepes-tris (pH 7.5) with osmolality of 540 mmol/kg (29). Fresh 5 mM Mg·ATP and 5 mM tris-GTP were added daily (29). When we studied the role of pH in anion channel regulation by ABA the pH buffer in the pipette solution was 0.1 mM Hepes-tris (pH 7.5). The bath solution was 30 mM CsCl 2 mM MgCl 2 5 mM CaCl 2 10 mM Mes-tris (pH 5.6) with osmolality of 560 mmol/kg. Protoplasts were pretreated for at least 1.5 hours with ABA and 50 μM ABA was added to bath and pipette solutions (29). Anion currents were recorded 11 to 14 min after achieving the whole-cell configuration. Steady-state currents were averaged between 52.5 and 62.5 s.
  32. 10.1093/jxb/48.Special_Issue.515
  33. 10.1002/j.1460-2075.1990.tb07608.x
  34. 10.1007/BF00195690
  35. For Fig. 3C stomatal closure was assayed according to (29) except that after ABA application (20 μM) leaves were incubated for 3 hours before aperture measurement. For Fig. 3F we assayed stomatal closure with the protocol of (34) although with minor modification. Leaves were incubated for 2 hours under 450 μmol m −2 s −1 of light followed by the addition of 1 mM Na-butyrate and 20 μM ABA. Apertures were measured 2 hours later. The wild-type butyrate response (48) was saturated with 1 mM Na-butyrate.
  36. 10.1073/pnas.89.5.1790
  37. Leung J., Merlot S., Giraudat J., Plant Cell 9, 759 (1997). / Plant Cell by Leung J. (1997)
  38. On the basis of (37) 5 to 6 young leaves were excised from an individual plant and their total fresh weight was measured during incubation in a chamber through which dry air was passed.
  39. 10.2307/3869164
  40. 10.1073/pnas.91.14.6310
  41. 10.1046/j.1365-313X.1995.08020187.x
  42. 10.1104/pp.102.1.95
  43. 10.1046/j.1365-313X.1995.8040479.x
  44. 10.1105/tpc.11.12.2283
  45. 10.1073/pnas.88.20.8925
  46. 10.1104/pp.113.1.269
  47. 10.1073/pnas.97.21.11638
  48. X.-Q. Wang S. M. Assmann unpublished observations.
  49. Supported by USDA grants 98-35304-6681 and 2001-35304-09916 and NSF grant MCB-9874438 to S.M.A. We thank Z.-M. Pei for advice on patch-clamping Arabidopsis guard cells.
Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 5:52 a.m.)
Deposited 1 year, 7 months ago (Jan. 9, 2024, 5:28 p.m.)
Indexed 1 week, 1 day ago (Aug. 29, 2025, 6:26 a.m.)
Issued 24 years, 2 months ago (June 15, 2001)
Published 24 years, 2 months ago (June 15, 2001)
Published Print 24 years, 2 months ago (June 15, 2001)
Funders 0

None

@article{Wang_2001, title={G Protein Regulation of Ion Channels and Abscisic Acid Signaling in Arabidopsis Guard Cells}, volume={292}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1059046}, DOI={10.1126/science.1059046}, number={5524}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Wang, Xi-Qing and Ullah, Hemayet and Jones, Alan M. and Assmann, Sarah M.}, year={2001}, month=jun, pages={2070–2072} }