Sexual Plant Reproduction

, Volume 24, Issue 4, pp 319–326 | Cite as

Misregulation of phosphoinositides in Arabidopsis thaliana decreases pollen hydration and maternal fertility

Short Communication


Phosphoinositides are important lipids involved in membrane identity, vesicle trafficking, and intracellular signaling. In recent years, phosphoinositides have been shown to play a critical role in polarized secretion in plants, as perturbations of phosphoinositide metabolism, through loss of function mutants, result in defects in root hair elongation and pollen tube growth, where polarized secretion occurs rapidly. In the Brassicaceae, responses of stigmatic papillae to compatible pollen are also thought to involve highly regulated secretory events to facilitate pollen hydration and penetration of the pollen tube through the stigmatic surface. We therefore sought to analyze the female sporophyte fertility of the root hair defective4-1 mutant and the PI 4-kinase β1/β2 double mutant, which differentially affect phosphatidylinositol-4-phosphate (PI4P) levels. Stigmas from both mutants supported slower rates of pollen grain hydration, and the fecundity of these mutants was also diminished as a result of failed pollination events. This study therefore concludes that PI4P is integral to appropriate pistil responses to compatible pollen.


Pollen-pistil interactions Compatible pollen PI 4-kinase Phosphoinositide phosphatase Arabidopsis 



We are very grateful to Dr. Erik Nielsen for helpful discussions and for providing the rhd4-1 and pi4kβ1/β2 mutants used in this study. We also thank Emily Indriolo for critical reading of the manuscript. LAC was supported by an Ontario Graduate Scholarship and a graduate scholarship from the Natural Sciences and Engineering Research Council of Canada (NSERC). Research in the laboratory of DRG is supported by grants from NSERC and a Canada Research Chair to DRG.


  1. Abramoff MD, Magelhaes PJ, Ram SJ (2004) Image processing with Image. J Biophotonics Int 11:36–42Google Scholar
  2. Audhya A, Emr SD (2002) Stt4 PI 4-kinase localizes to the plasma membrane and functions in the Pkc1-mediated MAP kinase cascade. Dev Cell 2:593–605PubMedCrossRefGoogle Scholar
  3. Chapman LA, Goring DR (2010) Pollen-pistil interactions regulating successful fertilization in the Brassicaceae. J Exp Bot 61:1987–1999PubMedCrossRefGoogle Scholar
  4. Despres B, Bouissonnie F, Wu HJ, Gomord V, Guilleminot J, Grellet F, Berger F, Delseny M, Devic M (2003) Three SAC1-like genes show overlapping patterns of expression in Arabidopsis but are remarkably silent during embryo development. Plant J 34:293–306PubMedCrossRefGoogle Scholar
  5. Elleman CJ, Dickinson HG (1986) Pollen stigma interactions in Brassica. IV. Structural reorganization in the pollen grains during hydration. J Cell Sci 80:141–157PubMedGoogle Scholar
  6. Elleman CJ, Franklin-Tong V, Dickinson HG (1992) Pollination in species with dry stigmas: the nature of the early stigmatic response and the pathway taken by pollen tubes. New Phytol 121:413–424CrossRefGoogle Scholar
  7. Fendrych M, Synek L, Pecenková T, Toupalová H, Cole R, Drdová E, Nebesárová J, Sedinová M, Hála M, Fowler JE, Zársky V (2010) The Arabidopsis exocyst complex is involved in cytokinesis and cell plate maturation. Plant Cell 22:3053–3065PubMedCrossRefGoogle Scholar
  8. Galvao RM, Kota U, Soderblom EJ, Goshe MB, Boss WF (2008) Characterization of a new family of protein kinases from Arabidopsis containing phosphoinositide 3/4-kinase and ubiquitin-like domains. Biochem J 409:117–127PubMedCrossRefGoogle Scholar
  9. Gillaspy GE (2010) Signaling and the polyphosphoinositide phosphatases from plants. In: Munnik T (ed) Lipid signaling in plants. Springer, Berlin, pp 117–130CrossRefGoogle Scholar
  10. Hála M, Cole R, Synek L, Drdová E, Pecenková T, Nordheim A, Lamkemeyer T, Madlung J, Hochholdinger F, Fowler JE, Zárský V (2008) An exocyst complex functions in plant cell growth in Arabidopsis and tobacco. Plant Cell 20:1330–1345PubMedCrossRefGoogle Scholar
  11. He B, Guo W (2009) The exocyst complex in polarized exocytosis. Curr Opin Cell Biol 21:537–542PubMedCrossRefGoogle Scholar
  12. Hiscock SJ, Allen AM (2008) Diverse cell signalling pathways regulate pollen-stigma interactions: the search for consensus. New Phytol 179:286–317PubMedCrossRefGoogle Scholar
  13. Iwano M, Shiba H, Miwa T, Che FS, Takayama S, Nagai T, Miyawaki A, Isogai A (2004) Ca2+dynamics in a pollen grain and papilla cell during pollination of Arabidopsis. Plant Physiol 136:3562–3571PubMedCrossRefGoogle Scholar
  14. Kandasamy MK, Nasrallah JB, Nasrallah ME (1994) Pollen–pistil interactions and developmental regulation of pollen tube growth in Arabidopsis. Development 120:405–418Google Scholar
  15. Kang BH, Nielsen E, Preuss ML, Mastronarde D, Staehelin LA (2011) Electron tomography of RabA4b- and PI-4Kbeta1-labeled trans Golgi network compartments in Arabidopsis. Traffic 12:313–329PubMedCrossRefGoogle Scholar
  16. Konrad G, Schlecker T, Faulhammer F, Mayinger P (2002) Retention of the yeast Sac1p phosphatase in the endoplasmic reticulum causes distinct changes in cellular phosphoinositide levels and stimulates microsomal ATP transport. J Biol Chem 277:10547–10554PubMedCrossRefGoogle Scholar
  17. Kost B, Lemichez E, Spielhofer P, Hong Y, Tolias K, Carpenter C, Chua NH (1999) Rac homologues and compartmentalized phosphatidylinositol 4, 5-bisphosphate act in a common pathway to regulate polar pollen tube growth. J Cell Biol 145:317–330PubMedCrossRefGoogle Scholar
  18. Kulich I, Cole R, Drdová E, Cvrcková F, Soukup A, Fowler J, Zárský V (2010) Arabidopsis exocyst subunits SEC8 and EXO70A1 and exocyst interactor ROH1 are involved in the localized deposition of seed coat pectin. New Phytol 188:615–625PubMedCrossRefGoogle Scholar
  19. Mayfield JA, Preuss D (2000) Rapid initiation of Arabidopsis pollination requires the oleosin-domain protein GRP17. Nat Cell Biol 2:128–130PubMedCrossRefGoogle Scholar
  20. Meinke DW, Sussex IM (1979) Embryo-lethal mutants of Arabidopsis thaliana. A model system for genetic analysis of plant embryo development. Dev Biol 72:50–61PubMedCrossRefGoogle Scholar
  21. Mueller-Roeber B, Pical C (2002) Inositol phospholipid metabolism in Arabidopsis. Characterized and putative isoforms of inositol phospholipid kinase and phosphoinositide-specific phospholipase C. Plant Physiol 130:22–46PubMedCrossRefGoogle Scholar
  22. Preuss ML, Serna J, Falbel TG, Bednarek SY, Nielsen E (2004) The Arabidopsis Rab GTPase RabA4b localizes to the tips of growing root hair cells. Plant Cell 16:1589–1603PubMedCrossRefGoogle Scholar
  23. Preuss ML, Schmitz AJ, Thole JM, Bonner HK, Otegui MS, Nielsen E (2006) A role for the RabA4b effector protein PI-4Kbeta1 in polarized expansion of root hair cells in Arabidopsis thaliana. J Cell Biol 172:991–998PubMedCrossRefGoogle Scholar
  24. Rhee SY, Beavis W, Berardini TZ, Chen GH, Dixon D, Doyle A, Garcia-Hernandez M, Huala E, Lander G, Montoya M, Miller N, Mueller LA, Mundodi S, Reiser L, Tacklind J, Weems DC, Wu YH, Xu I, Yoo D, Yoon J, Zhang PF (2003) The Arabidopsis information resource (TAIR): a model organism database providing a centralized, curated gateway to Arabidopsis biology, research materials and community. Nucleic Acids Res 31:224–228PubMedCrossRefGoogle Scholar
  25. Samuel MA, Chong YT, Haasen KE, Aldea-Brydges MG, Stone SL, Goring DR (2009) Cellular pathways regulating responses to compatible and self-incompatible pollen in Brassica and Arabidopsis stigmas intersect at Exo70A1, a putative component of the exocyst complex. Plant Cell 21:2655–2671PubMedCrossRefGoogle Scholar
  26. Schmid M, Davison TS, Henz SR, Pape UJ, Demar M, Vingron M, Scholkopf B, Weigel D, Lohmann JU (2005) A gene expression map of Arabidopsis thaliana development. Nature Genet 37:501–506PubMedCrossRefGoogle Scholar
  27. Stenzel I, Ischebeck T, Konig S, Holubowska A, Sporysz M, Hause B, Heilmann I (2008) The type B phosphatidylinositol-4-phosphate 5-kinase 3 is essential for root hair formation in Arabidopsis thaliana. Plant Cell 20:124–141PubMedCrossRefGoogle Scholar
  28. Stevenson JM, Perera IY, Boss WF (1998) A phosphatidylinositol 4-kinase pleckstrin homology domain that binds phosphatidylinositol 4-monophosphate. J Biol Chem 273:22761–22767PubMedCrossRefGoogle Scholar
  29. Stevenson-Paulik J, Love J, Boss WF (2003) Differential regulation of two Arabidopsis type III phosphatidylinositol 4-kinase isoforms. A regulatory role for the pleckstrin homology domain. Plant Physiol 132:1053–1064PubMedCrossRefGoogle Scholar
  30. Swanson R, Clark T, Preuss D (2005) Expression profiling of Arabidopsis stigma tissue identifies stigma-specific genes. Sex Plant Reprod 18:163–171CrossRefGoogle Scholar
  31. Synek L, Schlager N, Eliás M, Quentin M, Hauser MT, Zárský V (2006) AtEXO70A1, a member of a family of putative exocyst subunits specifically expanded in land plants, is important for polar growth and plant development. Plant J 48:54–72PubMedCrossRefGoogle Scholar
  32. Thole JM, Nielsen E (2008) Phosphoinositides in plants: novel functions in membrane trafficking. Curr Opin Plant Biol 11:620–631PubMedCrossRefGoogle Scholar
  33. Thole JM, Vermeer JE, Zhang Y, Gadella TW Jr, Nielsen E (2008) Root hair defective4 encodes a phosphatidylinositol-4-phosphate phosphatase required for proper root hair development in Arabidopsis thaliana. Plant Cell 20:381–395PubMedCrossRefGoogle Scholar
  34. Toufighi K, Brady SM, Austin R, Ly E, Provart NJ (2005) The botany array resource: e-Northerns, expression angling, and promoter analyses. Plant J 43:153–163PubMedCrossRefGoogle Scholar
  35. Tung CW, Dwyer KG, Nasrallah ME, Nasrallah JB (2005) Genome-wide identification of genes expressed in Arabidopsis pistils specifically along the path of pollen tube growth. Plant Physiol 138:977–989PubMedCrossRefGoogle Scholar
  36. Updegraff EP, Zhao F, Preuss D (2009) The extracellular lipase EXL4 is required for efficient hydration of Arabidopsis pollen. Sex Plant Reprod 22:197–204PubMedCrossRefGoogle Scholar
  37. Vermeer JE, Thole JM, Goedhart J, Nielsen E, Munnik T, Gadella TW Jr (2009) Imaging phosphatidylinositol 4-phosphate dynamics in living plant cells. Plant J 57:356–372PubMedCrossRefGoogle Scholar
  38. Williams ME, Torabinejad J, Cohick E, Parker K, Drake EJ, Thompson JE, Hortter M, Dewald DB (2005) Mutations in the Arabidopsis phosphoinositide phosphatase gene SAC9 lead to overaccumulation of PtdIns(4, 5)P2 and constitutive expression of the stress-response pathway. Plant Physiol 138:686–700PubMedCrossRefGoogle Scholar
  39. Winter D, Vinegar B, Nahal H, Ammar R, Wilson GV, Provart NJ (2007) An electronic fluorescent pictograph browser for exploring and analyzing large-scale biological data sets. PLoS One 2:e718(1–12)Google Scholar
  40. Xue HW, Pical C, Brearley C, Elge S, Muller-Rober B (1999) A plant 126-kDa phosphatidylinositol 4-kinase with a novel repeat structure. Cloning and functional expression in baculovirus-infected insect cells. J Biol Chem 274:5738–5745PubMedCrossRefGoogle Scholar
  41. Zárský V, Cvrcková F, Potocký M, Hála M (2009) Exocytosis and cell polarity in plants—exocyst and recycling domains. New Phytol 183:255–272PubMedCrossRefGoogle Scholar
  42. Zhang Y, Liu CM, Emons AM, Ketelaar T (2010) The plant exocyst. J Integr Plant Biol 52:138–146PubMedCrossRefGoogle Scholar
  43. Zhao Y, Yan A, Feijo J, Furutani M, Takenawa T, Hwang I, Fu Y, Yang Z (2010) Phosphoinositides regulate clathrin-dependent endocytosis at the tip of pollen tubes in Arabidopsis and tobacco. Plant Cell 22:4031–4044PubMedCrossRefGoogle Scholar
  44. Zhong R, Ye ZH (2003) The SAC domain-containing protein gene family in Arabidopsis. Plant Physiol 132:544–555PubMedCrossRefGoogle Scholar
  45. Zhong R, Burk DH, Nairn CJ, Wood-Jones A, Morrison WH 3rd, Ye ZH (2005) Mutation of SAC1, an Arabidopsis SAC domain phosphoinositide phosphatase, causes alterations in cell morphogenesis, cell wall synthesis, and actin organization. Plant Cell 17:1449–1466PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Cell & Systems BiologyUniversity of TorontoTorontoCanada
  2. 2.Centre for the Analysis of Genome Evolution and FunctionUniversity of TorontoTorontoCanada

Personalised recommendations