Home LiteratureArticle Details
PMID: 19114538 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Functional proteomics of Arabidopsis thaliana guard cells uncovers new stomatal signaling pathways.

The Plant cell ·Vol. 20 ·No. 12 ·2008-12-00 ·Pages 3210-26

Zhao Z, Zhang W, Stanley BA, Assmann SM

Abstract

We isolated a total of 3 x 10(8) guard cell protoplasts from 22,000 Arabidopsis thaliana plants and identified 1734 unique proteins using three complementary proteomic methods: protein spot identification from broad and narrow pH range two-dimensional (2D) gels, and 2D liquid chromatography-matrix assisted laser desorption/ionization multidimensional protein identification technology. This extensive single-cell-type proteome includes 336 proteins not previously represented in transcriptome analyses of guard cells and 52 proteins classified as signaling proteins by Gene Ontology analysis, of which only two have been previously assessed in the context of guard cell function. THIOGLUCOSIDE GLUCOHYDROLASE1 (TGG1), a myrosinase that catalyzes the production of toxic isothiocyanates from glucosinolates, showed striking abundance in the guard cell proteome. tgg1 mutants were hyposensitive to abscisic acid (ABA) inhibition of guard cell inward K(+) channels and stomatal opening, revealing that the glucosinolate-myrosinase system, previously identified as a defense against biotic invaders, is required for key ABA responses of guard cells. Our results also suggest a mechanism whereby exposure to abiotic stresses may enhance plant defense against subsequent biotic stressors and exemplify how enhanced knowledge of the signaling networks of a specific cell type can be gained by proteomics approaches.

MeSH Terms
Arabidopsis/genetics,metabolism Arabidopsis Proteins/genetics,metabolism,physiology Chromatography, Liquid Electrophoresis, Gel, Two-Dimensional Glycoside Hydrolases/genetics,metabolism,physiology Plant Stomata/cytology,genetics Proteome/analysis Signal Transduction/genetics,physiology Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Chemicals
Arabidopsis Proteins Proteome Glycoside Hydrolases TGG1 protein, Arabidopsis
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zhao Zhixin
Biology Department, Pen State University, University Park, Pensylvania 16802, USA.
Zhang Wei
Stanley Bruce A
Assmann Sarah M
References (111)
111 references, click to expand
  1. Roles of ion channels and transporters in guard cell signal transduction.
    FEBS Lett. 2007 May 25;581(12):2325-36 PMID: 17462636
  2. The zinc-finger protein Zat12 plays a central role in reactive oxygen and abiotic stress signaling in Arabidopsis.
    Plant Physiol. 2005 Oct;139(2):847-56 PMID: 16183833
  3. Ribulosebisphosphate carboxylase activity and photosynthetic o(2) evolution rate in vicia guard-cell protoplasts.
    Plant Physiol. 1989 Oct;91(2):459-63 PMID: 16667053
  4. Abscisic acid regulation of guard-cell K+ and anion channels in Gbeta- and RGS-deficient Arabidopsis lines.
    Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8476-81 PMID: 18541915
  5. Arabidopsis myrosinases TGG1 and TGG2 have redundant function in glucosinolate breakdown and insect defense.
    Plant J. 2006 May;46(4):549-62 PMID: 16640593
  6. High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome.
    Plant Mol Biol. 2005 Mar;57(4):577-91 PMID: 15821981
  7. Simultaneous analysis of phytohormones, phytotoxins, and volatile organic compounds in plants.
    Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10552-7 PMID: 12874387
  8. Mutants of Arabidopsis deficient in the synthesis of alpha-linolenate. Biochemical and genetic characterization of the endoplasmic reticulum linoleoyl desaturase.
    J Biol Chem. 1993 Aug 5;268(22):16345-51 PMID: 8102138
  9. Abscisic acid signal transduction in guard cells is mediated by phospholipase D activity.
    Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):12192-7 PMID: 10518598
  10. Regulation of abscisic acid-induced stomatal closure and anion channels by guard cell AAPK kinase.
    Science. 2000 Jan 14;287(5451):300-3 PMID: 10634783
  11. An Abscisic Acid-Activated and Calcium-Independent Protein Kinase from Guard Cells of Fava Bean.
    Plant Cell. 1996 Dec;8(12):2359-2368 PMID: 12239380
  12. Proteomics studies of post-translational modifications in plants.
    J Exp Bot. 2006;57(7):1547-51 PMID: 16551683
  13. Phospholipase D in the signaling networks of plant response to abscisic acid and reactive oxygen species.
    Biochim Biophys Acta. 2005 Sep 5;1736(1):1-9 PMID: 16099207
  14. The Paragon Algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra.
    Mol Cell Proteomics. 2007 Sep;6(9):1638-55 PMID: 17533153
  15. Regional vegetation die-off in response to global-change-type drought.
    Proc Natl Acad Sci U S A. 2005 Oct 18;102(42):15144-8 PMID: 16217022
  16. Patterns of protein oxidation in Arabidopsis seeds and during germination.
    Plant Physiol. 2005 Jun;138(2):790-802 PMID: 15908592
  17. A reference map of the Arabidopsis thaliana mature pollen proteome.
    Biochem Biophys Res Commun. 2005 Dec 2;337(4):1257-66 PMID: 16242667
  18. Arabidopsis thaliana MTP1 is a Zn transporter in the vacuolar membrane which mediates Zn detoxification and drives leaf Zn accumulation.
    FEBS Lett. 2005 Aug 1;579(19):4165-74 PMID: 16038907
  19. Proteomic analysis of soybean root hairs after infection by Bradyrhizobium japonicum.
    Mol Plant Microbe Interact. 2005 May;18(5):458-67 PMID: 15915644
  20. Genome-scale proteomics reveals Arabidopsis thaliana gene models and proteome dynamics.
    Science. 2008 May 16;320(5878):938-41 PMID: 18436743
  21. Proteome mapping of mature pollen of Arabidopsis thaliana.
    Proteomics. 2005 Dec;5(18):4864-84 PMID: 16247729
  22. Photosynthesis and nitrogen relationships in leaves of C3 plants.
    Oecologia. 1989 Jan;78(1):9-19 PMID: 28311896
  23. Ecosystem service supply and vulnerability to global change in Europe.
    Science. 2005 Nov 25;310(5752):1333-7 PMID: 16254151
  24. The plant innate immunity response in stomatal guard cells invokes G-protein-dependent ion channel regulation.
    Plant J. 2008 Dec;56(6):984-96 PMID: 18702674
  25. Proteomic analysis of different mutant genotypes of Arabidopsis led to the identification of 11 proteins correlating with adventitious root development.
    Plant Physiol. 2006 Jan;140(1):349-64 PMID: 16377752
  26. Blue-light- and phosphorylation-dependent binding of a 14-3-3 protein to phototropins in stomatal guard cells of broad bean.
    Plant Physiol. 2003 Dec;133(4):1453-63 PMID: 14605223
  27. Within-plant signalling via volatiles overcomes vascular constraints on systemic signalling and primes responses against herbivores.
    Ecol Lett. 2007 Jun;10(6):490-8 PMID: 17498148
  28. Ca(2+) signalling and control of guard-cell volume in stomatal movements.
    Curr Opin Plant Biol. 2000 Jun;3(3):196-204 PMID: 10837261
  29. Plant proteome analysis: a 2006 update.
    Proteomics. 2007 Aug;7(16):2947-62 PMID: 17654459
  30. Arabidopsis thaliana DNA gyrase is targeted to chloroplasts and mitochondria.
    Proc Natl Acad Sci U S A. 2004 May 18;101(20):7821-6 PMID: 15136745
  31. Phospholipase C is required for the control of stomatal aperture by ABA.
    Plant J. 2003 Apr;34(1):47-55 PMID: 12662308
  32. Glucosinolate genetics and the attraction of the aphid parasitoid Diaeretiella rapae to Brassica.
    Proc Biol Sci. 2000 Jan 7;267(1438):89-95 PMID: 10670958
  33. A proteome-wide protein interaction map for Campylobacter jejuni.
    Genome Biol. 2007;8(7):R130 PMID: 17615063
  34. Improved scoring of functional groups from gene expression data by decorrelating GO graph structure.
    Bioinformatics. 2006 Jul 1;22(13):1600-7 PMID: 16606683
  35. A proteome approach defines protective functions of tobacco leaf trichomes.
    Proteomics. 2005 Jul;5(10):2508-18 PMID: 15984042
  36. Voltage dependence of K channels in guard-cell protoplasts.
    Proc Natl Acad Sci U S A. 1987 Jun;84(12):4108-12 PMID: 16593851
  37. A gene expression map of the Arabidopsis root.
    Science. 2003 Dec 12;302(5652):1956-60 PMID: 14671301
  38. CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca(2+)-permeable channels and stomatal closure.
    PLoS Biol. 2006 Oct;4(10):e327 PMID: 17032064
  39. An internal rearrangement in an Arabidopsis inverted repeat locus impairs DNA methylation triggered by the locus.
    Genetics. 2004 Jan;166(1):437-48 PMID: 15020434
  40. Protein phosphorylation is a prerequisite for intracellular Ca2+ release and ion channel control by nitric oxide and abscisic acid in guard cells.
    Plant J. 2005 Aug;43(4):520-9 PMID: 16098106
  41. Cell identity mediates the response of Arabidopsis roots to abiotic stress.
    Science. 2008 May 16;320(5878):942-5 PMID: 18436742
  42. Chloroplast biogenesis: the use of mutants to study the etioplast-chloroplast transition.
    Proc Natl Acad Sci U S A. 2007 Jan 9;104(2):678-83 PMID: 17202255
  43. Drought-induced guard cell signal transduction involves sphingosine-1-phosphate.
    Nature. 2001 Mar 29;410(6828):596-9 PMID: 11279499
  44. From genomics to proteomics.
    Nature. 2003 Mar 13;422(6928):193-7 PMID: 12634792
  45. Calcium channels activated by hydrogen peroxide mediate abscisic acid signalling in guard cells.
    Nature. 2000 Aug 17;406(6797):731-4 PMID: 10963598
  46. Cytoplasmic alkalization precedes reactive oxygen species production during methyl jasmonate- and abscisic acid-induced stomatal closure.
    Plant Physiol. 2004 Apr;134(4):1536-45 PMID: 15064385
  47. Guard cell ABA and CO2 signaling network updates and Ca2+ sensor priming hypothesis.
    Curr Opin Plant Biol. 2006 Dec;9(6):654-63 PMID: 17010657
  48. 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.
    Plant Physiol. 2007 Mar;143(3):1398-407 PMID: 17220365
  49. Identification of a flavin-monooxygenase as the S-oxygenating enzyme in aliphatic glucosinolate biosynthesis in Arabidopsis.
    Plant J. 2007 Jun;50(5):902-10 PMID: 17461789
  50. Stress-induced protein S-glutathionylation in Arabidopsis.
    Plant Physiol. 2005 Aug;138(4):2233-44 PMID: 16055689
  51. Projected increase in continental runoff due to plant responses to increasing carbon dioxide.
    Nature. 2007 Aug 30;448(7157):1037-41 PMID: 17728755
  52. Shotgun proteomic analysis of Arabidopsis thaliana leaves.
    J Sep Sci. 2007 Sep;30(14):2225-30 PMID: 17654619
  53. Arabidopsis cell wall proteome defined using multidimensional protein identification technology.
    Proteomics. 2006 Jan;6(1):301-11 PMID: 16287169
  54. Inhibition of inward K+ channels and stomatal response by abscisic acid: an intracellular locus of phytohormone action.
    Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):4019-23 PMID: 8171028
  55. High resolution X-ray crystallography shows that ascorbate is a cofactor for myrosinase and substitutes for the function of the catalytic base.
    J Biol Chem. 2000 Dec 15;275(50):39385-93 PMID: 10978344
  56. Plant stomata function in innate immunity against bacterial invasion.
    Cell. 2006 Sep 8;126(5):969-80 PMID: 16959575
  57. TRP channel activation by reversible covalent modification.
    Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19564-8 PMID: 17164327
  58. Proteomic analysis of rice seedlings during cold stress.
    Proteomics. 2007 Apr;7(8):1293-302 PMID: 17380535
  59. Regulation of plant glucosinolate metabolism.
    Planta. 2007 Nov;226(6):1343-52 PMID: 17899172
  60. The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein alpha subunit GPA1 and regulates abscisic acid signaling.
    Plant Cell. 2004 Jun;16(6):1616-32 PMID: 15155892
  61. Hormone interactions in stomatal function.
    Plant Mol Biol. 2009 Mar;69(4):451-62 PMID: 19031047
  62. Metabolic energy for stomatal opening. Roles of photophosphorylation and oxidative phosphorylation.
    Planta. 1984 May;161(2):129-36 PMID: 24253600
  63. Variation of glucosinolate accumulation among different organs and developmental stages of Arabidopsis thaliana.
    Phytochemistry. 2003 Feb;62(3):471-81 PMID: 12620360
  64. Single cell proteomics for personalised medicine.
    Trends Mol Med. 2004 Dec;10(12):574-7 PMID: 15567324
  65. Gel-free mass spectrometry-based high throughput proteomics: tools for studying biological response of proteins and proteomes.
    Proteomics. 2006 Sep;6(17):4678-87 PMID: 16888762
  66. Microarray expression analyses of Arabidopsis guard cells and isolation of a recessive abscisic acid hypersensitive protein phosphatase 2C mutant.
    Plant Cell. 2004 Mar;16(3):596-615 PMID: 14973164
  67. G protein regulation of ion channels and abscisic acid signaling in Arabidopsis guard cells.
    Science. 2001 Jun 15;292(5524):2070-2 PMID: 11408655
  68. Glucosinolate metabolism and its control.
    Trends Plant Sci. 2006 Feb;11(2):89-100 PMID: 16406306
  69. The effect of alpha-amanitin on the Arabidopsis seed proteome highlights the distinct roles of stored and neosynthesized mRNAs during germination.
    Plant Physiol. 2004 Apr;134(4):1598-613 PMID: 15047896
  70. Before gene expression: early events in plant-insect interaction.
    Trends Plant Sci. 2007 Jul;12(7):310-6 PMID: 17596996
  71. Digging deeper into the plant cell wall proteome.
    Plant Physiol Biochem. 2004 Dec;42(12):979-88 PMID: 15707835
  72. Light regulation of stomatal movement.
    Annu Rev Plant Biol. 2007;58:219-47 PMID: 17209798
  73. Volatile signaling in plant-plant interactions: "talking trees" in the genomics era.
    Science. 2006 Feb 10;311(5762):812-5 PMID: 16469918
  74. Proteomics: from gel based to gel free.
    Anal Chem. 2005 Jun 15;77(12):3771-87 PMID: 15952756
  75. In-depth analysis of the thylakoid membrane proteome of Arabidopsis thaliana chloroplasts: new proteins, new functions, and a plastid proteome database.
    Plant Cell. 2004 Feb;16(2):478-99 PMID: 14729914
  76. Sealing plant surfaces: cuticular wax formation by epidermal cells.
    Annu Rev Plant Biol. 2008;59:683-707 PMID: 18251711
  77. Deep coverage mouse red blood cell proteome: a first comparison with the human red blood cell.
    Mol Cell Proteomics. 2008 Jul;7(7):1317-30 PMID: 18344233
  78. Profiling core proteomes of human cell lines by one-dimensional PAGE and liquid chromatography-tandem mass spectrometry.
    Mol Cell Proteomics. 2003 Dec;2(12):1297-305 PMID: 14532353
  79. AtVAM3 is required for normal specification of idioblasts, myrosin cells.
    Plant Cell Physiol. 2006 Jan;47(1):164-75 PMID: 16306062
  80. The Arabidopsis thaliana chloroplast proteome reveals pathway abundance and novel protein functions.
    Curr Biol. 2004 Mar 9;14(5):354-62 PMID: 15028209
  81. Phosphatidylinositol 3- and 4-phosphate are required for normal stomatal movements.
    Plant Cell. 2002 Oct;14(10):2399-412 PMID: 12368494
  82. Phosphorylation and Dephosphorylation of Guard-Cell Proteins from Vicia faba L. in Response to Light and Dark.
    Plant Physiol. 1993 Jul;102(3):917-923 PMID: 12231877
  83. Analysis of the Arabidopsis cytosolic ribosome proteome provides detailed insights into its components and their post-translational modification.
    Mol Cell Proteomics. 2008 Feb;7(2):347-69 PMID: 17934214
  84. Predicting essential components of signal transduction networks: a dynamic model of guard cell abscisic acid signaling.
    PLoS Biol. 2006 Oct;4(10):e312 PMID: 16968132
  85. Histone H2A.Z and homologues of components of the SWR1 complex are required to control immunity in Arabidopsis.
    Plant J. 2008 Feb;53(3):475-87 PMID: 17988222
  86. Five-group distribution of the Shaker-like K+ channel family in higher plants.
    J Mol Evol. 2003 Apr;56(4):418-34 PMID: 12664162
  87. In-depth analysis of the membrane and cytosolic proteome of red blood cells.
    Blood. 2006 Aug 1;108(3):791-801 PMID: 16861337
  88. Sphingolipid signalling in Arabidopsis guard cells involves heterotrimeric G proteins.
    Nature. 2003 Jun 5;423(6940):651-4 PMID: 12789341
  89. The role of stomata in sensing and driving environmental change.
    Nature. 2003 Aug 21;424(6951):901-8 PMID: 12931178
  90. The ectopically parting cells 1-2 (epc1-2) mutant exhibits an exaggerated response to abscisic acid.
    J Exp Bot. 2007;58(7):1813-23 PMID: 17426055
  91. Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool.
    Plant Methods. 2008 Feb 19;4:6 PMID: 18284694
  92. Arabidopsis OST1 protein kinase mediates the regulation of stomatal aperture by abscisic acid and acts upstream of reactive oxygen species production.
    Plant Cell. 2002 Dec;14(12):3089-99 PMID: 12468729
  93. The HIC signalling pathway links CO2 perception to stomatal development.
    Nature. 2000 Dec 7;408(6813):713-6 PMID: 11130071
  94. Composition and content of glucosinolates in developing Arabidopsis thaliana.
    Planta. 2002 Feb;214(4):562-71 PMID: 11925040
  95. Guard cells: a dynamic signaling model.
    Curr Opin Plant Biol. 2004 Oct;7(5):537-46 PMID: 15337096
  96. Biology and biochemistry of glucosinolates.
    Annu Rev Plant Biol. 2006;57:303-33 PMID: 16669764
  97. The vegetative vacuole proteome of Arabidopsis thaliana reveals predicted and unexpected proteins.
    Plant Cell. 2004 Dec;16(12):3285-303 PMID: 15539469
  98. Proteome dynamics during plastid differentiation in rice.
    Plant Physiol. 2007 Feb;143(2):912-23 PMID: 17189339
  99. Glucosinolate research in the Arabidopsis era.
    Trends Plant Sci. 2002 Jun;7(6):263-70 PMID: 12049923
  100. NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis.
    EMBO J. 2003 Jun 2;22(11):2623-33 PMID: 12773379
  101. The ascorbic acid redox state controls guard cell signaling and stomatal movement.
    Plant Cell. 2004 May;16(5):1143-62 PMID: 15084716
  102. The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins.
    Genome Biol. 2006;7(9):R80 PMID: 16948836
  103. A bifurcating pathway directs abscisic acid effects on stomatal closure and opening in Arabidopsis.
    Science. 2006 Apr 14;312(5771):264-6 PMID: 16614222
  104. Arabidopsis hapless mutations define essential gametophytic functions.
    Genetics. 2004 Oct;168(2):971-82 PMID: 15514068
  105. Mass spectrometry. An essential tool in proteome analysis.
    Plant Physiol. 2000 Jul;123(3):807-9 PMID: 10889229
  106. Death don't have no mercy and neither does calcium: Arabidopsis CYCLIC NUCLEOTIDE GATED CHANNEL2 and innate immunity.
    Plant Cell. 2007 Mar;19(3):1081-95 PMID: 17384171
  107. Priming for stress resistance: from the lab to the field.
    Curr Opin Plant Biol. 2007 Aug;10(4):425-31 PMID: 17644024
  108. Cell-specific protein profiling in Arabidopsis thaliana trichomes: identification of trichome-located proteins involved in sulfur metabolism and detoxification.
    Phytochemistry. 2004 Jun;65(11):1641-9 PMID: 15276459
  109. Abscisic Acid-Induced Phosphoinositide Turnover in Guard Cell Protoplasts of Vicia faba.
    Plant Physiol. 1996 Mar;110(3):987-996 PMID: 12226236
  110. Guard cell- and phloem idioblast-specific expression of thioglucoside glucohydrolase 1 (myrosinase) in Arabidopsis.
    Plant Physiol. 2002 Apr;128(4):1180-8 PMID: 11950967
  111. Experimental analysis of the Arabidopsis mitochondrial proteome highlights signaling and regulatory components, provides assessment of targeting prediction programs, and indicates plant-specific mitochondrial proteins.
    Plant Cell. 2004 Jan;16(1):241-56 PMID: 14671022
Article Info
Journal
The Plant cell
Abbr.
Plant Cell
ISSN
1040-4651
DOI
10.1105/tpc.108.063263
Published
2008-12-00
Epub
2008-00-29
Pages
3210-26
Language
English
Region
England
NLM ID
9208688
PMCID
PMC2630442
Subset
IM
Corrections
CommentIn
PMID: 19114536
External Links
PubMed source DOI: 10.1105/tpc.108.063263 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2