Abstract
Key message
Gymnosperm pollen grains release ATP to the extracellular matrix which is essential for the initiation of pollen germination and tube growth.
Abstract
Extracellular ATP (eATP) is an important signaling compound involved in various processes of animal and angiosperm cells. However, the role played by eATP in gymnosperm cells remains unclear. Using a bioluminescence assay, we found that pollen grains of Picea meyeri released ATP to the extracellular matrix before germination and during tube elongation. The addition of further exogenous ATP or an apyrase inhibitor to pollen suspensions inhibited germination and pollen tube elongation. Exogenous apyrase (which hydrolyzes eATP released from pollen per se) exerted a similar inhibitory effect. Moreover, incubation of pollen suspensions with purinoceptor inhibitors prevented germination. ATP intensified the influx of Ca2+ after germination, which was abrogated by purinoceptor inhibitors. Confocal microscopy revealed that the microfilament pattern became disorganized in pollen tubes when exposed to ATP. Together, our findings suggest that optimum concentration of eATP is essential for initiation of pollen germination, and eATP signaling regulates pollen tube growth by activating purinoceptors to increase Ca2+ influx, thus modulating microfilament organization, which, in turn, is essential for pollen germination and tube growth. Hence, we provide a mechanistic framework for the role played by eATP in pollen germination and tube growth.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Adler EM (2014) Of ATP receptors, opioid receptors, and AKAP regulation of calcium channels. J Gen Physiol 143:313–314
Chen T, Teng N, Wu X, Wang Y, Tang W, Samaj J, Baluska F, Lin J (2007) Disruption of actin filaments by latrunculin B affects cell wall construction in Picea meyeri pollen tube by disturbing vesicle trafficking. Plant Cell Physiol 48:19–30
Chen KM, Wu GL, Wang YH, Tian CT, Samaj J, Baluska F, Lin JX (2008) The block of intracellular calcium release affects the pollen tube development of Picea wilsonii by changing the deposition of cell wall components. Protoplasma 233:39–49
Chivasa S, Murphy AM, Hamilton JM, Lindsey K, Carr JP, Slabas AR (2009) Extracellular ATP is a regulator of pathogen defence in plants. Plant J 60:436–448
Choi J, Tanaka K, Cao Y, Qi Y, Qiu J, Liang Y, Lee SY, Stacey G (2014) Identification of a plant receptor for extracellular ATP. Science 343:290–294
Clark G, Roux SJ (2011) Apyrases, extracellular ATP and the regulation of growth. Curr Opin Plant Biol 14:700–706
Demidchik V, Shang Z, Shin R, Thompson E, Rubio L, Laohavisit A, Mortimer JC, Chivasa S, Slabas AR, Glover BJ, Schachtman DP, Shabala SN, Davies JM (2009) Plant extracellular ATP signalling by plasma membrane NADPH oxidase and Ca2+ channels. Plant J 58:903–913
Dutta AK, Okada Y, Sabirov RZ (2002) Regulation of an ATP-conductive large-conductance anion channel and swelling-induced ATP release by arachidonic acid. J Physiol 542:803–816
Franklin-Tong VE (1999) Signaling and the modulation of pollen tube growth. Plant Cell 11:727–738
Holdaway-Clarke TL, Feijo JA, Hackett GR, Kunkel JG, Hepler PK (1997) Pollen tube growth and the intracellular cytosolic calcium gradient oscillate in phase while extracellular calcium influx is delayed. Plant Cell 9:1999–2010
Khakh BS, Burnstock G (2009) The double life of ATP. Sci Am 301:84–92
Kim SY, Sivaguru M, Stacey G (2006) Extracellular ATP in plants. Visualization, localization, and analysis of physiological significance in growth and signaling. Plant Physiol 142:984–992
Krichevsky A, Kozlovsky SV, Tian GW, Chen MH, Zaltsman A, Citovsky V (2007) How pollen tubes grow. Dev Biol 303:405–420
Maloney SF, Brass LF, Diamond SL (2010) P2Y12 or P2Y1 inhibitors reduce platelet deposition in a microfluidic model of thrombosis while apyrase lacks efficacy under flow conditions. Integr Biol (Camb) 2:183–192
Mazurek S, Michel A, Eigenbrodt E (1997) Effect of extracellular AMP on cell proliferation and metabolism of breast cancer cell lines with high and low glycolytic rates. J Biol Chem 272:4941–4952
Ou GS, Chen ZL, Yuan M (2002) Jasplakinolide reversibly disrupts actin filaments in suspension-cultured tobacco BY-2 cells. Protoplasma 219:168–175
Ralevic V, Burnstock G (1998) Receptors for purines and pyrimidines. Pharmacol Rev 50:413–492
Reichler SA, Torres J, Rivera AL, Cintolesi VA, Clark G, Roux SJ (2009) Intersection of two signalling pathways: extracellular nucleotides regulate pollen germination and pollen tube growth via nitric oxide. J Exp Bot 60:2129–2138
Rieder B, Neuhaus HE (2011) Identification of an Arabidopsis plasma membrane-located ATP transporter important for anther development. Plant Cell 23:1932–1944
Rittiner JE, Korboukh I, Hull-Ryde EA, Jin J, Janzen WP, Frye SV, Zylka MJ (2012) AMP is an adenosine A1 receptor agonist. J Biol Chem 287:5301–5309
Shabir S, Cross W, Kirkwood LA, Pearson JF, Appleby PA, Walker D, Eardley I, Southgate J (2013) Functional expression of purinergic P2 receptors and transient receptor potential channels by the human urothelium. Am J Physiol Renal Physiol 305:F396–F406
Solini A, Iacobini C, Ricci C, Chiozzi P, Amadio L, Pricci F, Di Mario U, Di Virgilio F, Pugliese G (2005) Purinergic modulation of mesangial extracellular matrix production: role in diabetic and other glomerular diseases. Kidney Int 67:875–885
Steinebrunner I, Wu J, Sun Y, Corbett A, Roux SJ (2003) Disruption of apyrases inhibits pollen germination in Arabidopsis. Plant Physiol 131:1638–1647
Sun J, Zhang CL, Deng SR, Lu CF, Shen X, Zhou XY, Zheng XJ, Hu ZM, Chen SL (2012) An ATP signalling pathway in plant cells: extracellular ATP triggers programmed cell death in Populus euphratica. Plant Cell Environ 35:893–916
Tanaka K, Gilroy S, Jones AM, Stacey G (2010) Extracellular ATP signaling in plants. Trends Cell Biol 20:601–608
Vidali L, McKenna ST, Hepler PK (2001) Actin polymerization is essential for pollen tube growth. Mol Biol Cell 12:2534–2545
Wang Y, Chen T, Zhang C, Hao H, Liu P, Zheng M, Baluska F, Samaj J, Lin J (2009) Nitric oxide modulates the influx of extracellular Ca2+ and actin filament organization during cell wall construction in Pinus bungeana pollen tubes. New Phytol 182:851–862
Weerasinghe RR, Swanson SJ, Okada SF, Garrett MB, Kim SY, Stacey G, Boucher RC, Gilroy S, Jones AM (2009) Touch induces ATP release in Arabidopsis roots that is modulated by the heterotrimeric G-protein complex. FEBS Lett 583:2521–2526
Windsor JB, Thomas C, Hurley L, Roux SJ, Lloyd AM (2002) Automated colorimetric screen for apyrase inhibitors. Biotechniques 33:1024–1030
Wu J, Steinebrunner I, Sun Y, Butterfield T, Torres J, Arnold D, Gonzalez A, Jacob F, Reichler S, Roux SJ (2007) Apyrases (nucleoside triphosphate-diphosphohydrolases) play a key role in growth control in Arabidopsis. Plant Physiol 144:961–975
Yegutkin GG, Mikhailov A, Samburski SS, Jalkanen S (2006) The detection of micromolar pericellular ATP pool on lymphocyte surface by using lymphoid ecto-adenylate kinase as intrinsic ATP sensor. Mol Biol Cell 17:3378–3385
Yu W, Robson SC, Hill WG (2011) Expression and distribution of ectonucleotidases in mouse urinary bladder. PLoS ONE 6:e18704
Author contribution statement
Yanping Jing and Junhui Zhou designed the experiment, Junhui Zhou performed most of the experimental work, Kai Liu carried out ATP assay experiment, Junhui Zhou and Chengyu Fan wrote the paper. All authors read and approved the final manuscript.
Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (No. 31371348), Program for Changjiang Scholars and Innovative Research Team in University (No. IRT13047).
Conflict of interest
We declare that we have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by S. Chen.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplemental Figure 1. (A) Neither 0.5 mM ADP nor AMP significantly affected the pollen germination rate. (B) Neither 0.5 mM ADP nor AMP significantly affected pollen tube growth. (C) Pollen germination was similarly inhibited by 5.0 mM ADP and AMP. (D) Pollen tube growth was inhibited by 5.0 mM ADP and AMP, and the effect of AMP was stronger. Significant difference at P < 0.05 (*) or 0.01 (**).
Supplemental Figure 2. (A) Pollen germination was inhibited by 0.2 mM ATPγS. (B) Pollen tube growth was inhibited by 0.2 mM ATPγS. Significant difference at P < 0.05 (*) or 0.01 (**).
Supplemental Figure 3. Pollen grains were treated with various concentrations of suramin and observed at 24, 36 and 48 h of treatment. Bar: 500 μm. (A) Pollen grains cultured in normal culture medium. (B-G) Pollen grains were exposed to 0.01, 0.02, 0.05, 0.1, 0.2, or 1.0 mM suramin, respectively. (H) Pollen grains were treated with 5 mM ATP. (I–N) Pollen suspensions were mixed with 5 mM ATP and 0.01, 0.02, 0.05, 0.1, 0.2, or 1.0 mM suramin, respectively.
Supplemental Figure 4. Pollen grains were exposed to various concentrations of PPADS and observed at 24, 36, and 48 h of treatment. Bar: 100 μm. (A-1, A-2, A-3, A-4, A-5, and A-6) Pollen grains were exposed to 0.1, 0.2, 0.5, 1.0, 2.0, or 5.0 mM PPADS, respectively, and growth was inhibited to different extents. (B-1, B-2, B-3, B-4, B-5, and B-6) Pollen suspensions were mixed with 5 mM ATP and 0.1, 0.2, 0.5, 1.0, 2.0, or 5.0 mM PPADS, respectively. Germinated pollen grains were further inhibited by 5 mM ATP.
Supplemental Figure 5. Pollen grains were treated with various concentrations of PPADS and observed at 24, 36 and 48 h of treatment. Bar: 500 μm. (A) Pollen grains cultured in normal culture medium. (B-G) Pollen grains were exposed to 0.1, 0.2, 0.5, 1.0, 2.0, or 5.0 mM PPADS, respectively. (H) Pollen grains were treated with 5 mM ATP. (I–N) Pollen suspensions were mixed with 5 mM ATP and 0.1, 0.2, 0.5, 1.0, 2.0, or 5.0 mM PPADS, respectively.
Rights and permissions
About this article
Cite this article
Zhou, J., Fan, C., Liu, K. et al. Extracellular ATP is involved in the initiation of pollen germination and tube growth in Picea meyeri . Trees 29, 563–574 (2015). https://doi.org/10.1007/s00468-014-1135-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00468-014-1135-6