Plant Cell Reports

, Volume 33, Issue 2, pp 255–263 | Cite as

Apoplastic calmodulin promotes self-incompatibility pollen tube growth by enhancing calcium influx and reactive oxygen species concentration in Pyrus pyrifolia

  • Xueting Jiang
  • Yongbin Gao
  • Hongsheng Zhou
  • Jianqing Chen
  • Juyou Wu
  • Shaoling ZhangEmail author
Original Paper


Key message

This study indicated that Ca 2+ , ROS and actin filaments were involved with CaM in regulating pollen tube growth and providing a potential way for overcoming pear self-incompatibility.


Calmodulin (CaM) has been associated with various physiological and developmental processes in plants, including pollen tube growth. In this study, we showed that CaM regulated the pear pollen tube growth in a concentration-dependent bi-phasic response. Using a whole-cell patch-clamp configuration, we showed that apoplastic CaM induced a hyperpolarization-activated calcium ion (Ca2+) current, and anti-CaM largely inhibited this type of Ca2+ current. Moreover, upon anti-CaM treatment, the reactive oxygen species (ROS) concentration decreased and actin filaments depolymerized in the pollen tube. Interestingly, CaM could partially rescue the inhibition of self-incompatible pear pollen tube growth. This phenotype could be mediated by CaM-enhanced pollen plasma membrane Ca2+ current, tip-localized ROS concentration and stabilized actin filaments. These data indicated that Ca2+, ROS and actin filaments were involved with CaM in regulating pollen tube growth and provide a potential way for overcoming pear self-incompatibility.


Pollen tube Pear Calmodulin Self-incompatibility 



This work was supported by the National Natural Science of China (31,071,759, 31,230,063 and 31,272,119), Doctoral Fund of Ministry of Education of China (20120097120046), Fund for Independent Innovation of Agricultural Sciences in Jiangsu Province (CX(11)1013, CX(13)3,010), National Natural Science Foundation of Jiangsu Province (BK2011067, BK2012366), and the Fundamental Research Funds for the Central Universities (KYRC201201).

Supplementary material

299_2013_1526_MOESM1_ESM.pdf (223 kb)
Supplementary material 1 (PDF 211 kb)


  1. Biro RL, Daye S, Serlin BS, Terry ME, Datta N, Sopory SK, Roux SJ (1984) Characterization of oat calmodulin and radioimmunoassay of its subcellular distribution. Plant Physiol 75:382–386PubMedCentralPubMedCrossRefGoogle Scholar
  2. Boonburapong B, Buaboocha T (2007) Genome-wide identification and analyses of the rice calmodulin and related potential calcium sensor proteins. BMC Plant Biol 7:4PubMedCentralPubMedCrossRefGoogle Scholar
  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  4. Brown PH, Ho THD (1986) Barley aleurone layers secrete a nuclease in response to gibberellic acid. Plant Physiol 82:801–806PubMedCentralPubMedCrossRefGoogle Scholar
  5. Cardennas L, Lovy-Wheeler A, Kunkel JG, Hepler PK (2008) Pollen tube growth oscillations and intracellular calcium levels are reversibly modulated by actin polymerization. Plant Physiol 146:1611–1621CrossRefGoogle Scholar
  6. Del Duca S, Bregoli AM, Bergamini C, Serafini-Fracassini D (1997) Transglutaminase-catalyzed modification of cytoskeletal proteins by polyamines during the germination of Malus domestica pollen. Sex Plant Reprod 10:89–95CrossRefGoogle Scholar
  7. Del Duca S, Serafini-Fracassini D, Bonner P, Cresti M, Cai G (2009) Effects of post-translational modifications catalysed by pollen transglutaminase on the functional properties of microtubules and actin filaments. Biochem J 418:651–664PubMedCrossRefGoogle Scholar
  8. Demidchik V, Maathuis FJ (2007) Physiological roles of nonselective cation channels in plants: from salt stress to signalling and development. New Phytol 175:387–404PubMedCrossRefGoogle Scholar
  9. Dixit R, Nasrallah JB (2001) Recognizing self in the self-incompatibility response. Plant Physiol 125:105–108PubMedCentralPubMedCrossRefGoogle Scholar
  10. Dutta R, Robinson KR (2004) Identification and characterization of stretch-activated ion channels in pollen protoplasts. Plant Physiol 135:1398–1406PubMedCentralPubMedCrossRefGoogle Scholar
  11. Foreman J, Demidchik V, Bothwell JHF, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JDG, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446PubMedCrossRefGoogle Scholar
  12. Franklin-Tong VE (1999) Signaling and the modulation of pollen tube growth. Plant Cell 11:727–738PubMedCentralPubMedGoogle Scholar
  13. Hiratsuka S, Zhang SL, Nakagawa E, Kawai Y (2001) Selective inhibition of the growth of incompatible pollen tubes by S-protein in the Japanese pear. Sex Plant Reprod 13:209–215CrossRefGoogle Scholar
  14. Huang S, Blanchoin L, Chaudhry F, Franklin-tong VE, Staiger CJ (2004) A gelsolin-like protein from Papaver rhoeas pollen (PrABP80) stimulates calcium-regulated severing and depolymerization of actin filaments. J Biol Chem 279:23364–23375PubMedCrossRefGoogle Scholar
  15. Kwak JM, Mori IC, Pei Z-M, Leonhardt N, Torres MA, Dangl JL, Bloom RE, Bodde S, Jones JD, Schroeder JI (2003) NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis. EMBO J 22:2623–2633PubMedCrossRefGoogle Scholar
  16. Lenartowska M, Rodriguez-Garcia M, Bednarska E (2001) Calmodulin and calmodulin-like protein are involved in pollen–pistil interaction: immunocytochemical studies on Petunia hybrida Hort. Acta Biol Cracov 43:117–123Google Scholar
  17. Liu ZQ, Xu GH, Zhang SL (2007) Pyrus pyrifolia stylar S-RNase induces alterations in the actin cytoskeleton in self-pollen and tubes in vitro. Protoplasma 232:61–67PubMedCrossRefGoogle Scholar
  18. Ma LG, Sun DY (1997) The effects of extracellular calmodulin on initiation of Hippeastrum rutilum pollen germination and tube growth. Planta 202:336–340CrossRefGoogle Scholar
  19. Ma LG, Xu XD, Cui SJ, Sun DY (1998) The involvement of phosphoinositide signaling pathway in the initiatory effects of extracellular calmodulin on pollen germination and tube growth. Acta Phytophysiol Sin 24:196–200Google Scholar
  20. Ma LG, Xu XD, Cui SJ, Sun DY (1999) The presence of a heterotrimeric G protein and its role in signal transduction of extracellular calmodulin in pollen germination and tube growth. Plant Cell 11:1351–1364PubMedCentralPubMedGoogle Scholar
  21. Ma LG, Fan QS, Yu ZQ, Zhou HL, Zhang FS, Sun DY (2000) Does aluminum inhibit pollen germination via extracellular calmodulin? Plant Cell Physiol 41:372–376PubMedCrossRefGoogle Scholar
  22. Malhó R, Read ND, Trewavas AJ, Pais MS (1995) Calcium channel activity during pollen tube growth and reorientation. Plant Cell 7:1173–1184PubMedCentralPubMedGoogle Scholar
  23. McCormack E, Tsai YC, Braam J (2005) Handling calcium signaling: Arabidopsis CaMs and CMLs. Trends Plant Sci 10:383–389PubMedCrossRefGoogle Scholar
  24. Michard E, Lima PT, Borges F, Silva AC, Portes MT, Carvalho JE, Gilliham M, Liu L-H, Obermeyer G, Feijó JA (2011) Glutamate receptor-like genes form Ca2+ channels in pollen tubes and are regulated by pistil d-serine. Sci Signal 332:434–437Google Scholar
  25. Monshausen GB, Bibikova TN, Weisenseel MH, Gilroy S (2009) Ca2+ regulates reactive oxygen species production and pH during mechanosensing in Arabidopsis roots. Plant Cell 21:2341–2356PubMedCentralPubMedCrossRefGoogle Scholar
  26. Pei ZM, Murata Y, Benning G, Thomine S, Klüsener B, Allen GJ, Grill E, Schroeder JI (2000) Calcium channels activated by hydrogen peroxide mediate abscisic acid signaling in guard cells. Nature 406:731–734PubMedCrossRefGoogle Scholar
  27. Potocky M, Jones MA, Bezvoda R, Smirnoff N, Žárský V (2007) Reactive oxygen species produced by NADPH oxidase are involved in pollen tube growth. New Phytol 174:742–751PubMedCrossRefGoogle Scholar
  28. Qu HY, Shang ZL, Zhang SL, Liu LM, Wu JY (2007) Identification of hyperpolarization-activated calcium channels in apical pollen tubes of Pyrus pyrifolia. New Phytol 174:524–536PubMedCrossRefGoogle Scholar
  29. Shang ZL, Ma LG, Zhang HL, He RR, Wang XC, Cui SJ, Sun DY (2005) Ca2+ influx into lily pollen grains through a hyperpolarization-activated Ca2+-permeable channel which can be regulated by extracellular CaM. Plant Cell Physiol 46:598–608PubMedCrossRefGoogle Scholar
  30. Snowman BN, Kovar DR, Shevchenko G, Franklin-Tong VE, Staiger CJ (2002) Signal-mediated depolymerization of actin in pollen during the self-incompatibility response. Plant Cell 14:2613–2626PubMedCentralPubMedCrossRefGoogle Scholar
  31. Sun DY, Li HB, Cheng G (1994) Extracellular calmodulin accelerates the proliferation of suspension-cultured cells of Angelica dahurica. Plant Sci 99:1–8CrossRefGoogle Scholar
  32. Sun DY, Bian YQ, Zhao BH, Zhao LY, Yu XM, Duan SJ (1995) The effects of extracellular calmodulin on cell wall regeneration of protoplasts and cell division. Plant Cell Physiol 36:133–138Google Scholar
  33. Sun Y, Liu DL, Yu ZQ, Zhang Q, Bai J, Sun DY (2003) An apoplastic mechanism for short-term effects of rare earth elements at lower concentrations. Plant Cell Environ 26:887–896PubMedCrossRefGoogle Scholar
  34. Wang YF, Fan LM, Zhang WZ, Zhang W, Wu WH (2004) Ca2+-permeable channels in the plasma membrane of Arabidopsis pollen are regulated by actin microfilaments. Plant Physiol 136:3892–3904PubMedCentralPubMedCrossRefGoogle Scholar
  35. Wang CL, Xu GH, Jiang XT, Chen G, Wu J, Wu HQ, Zhang SL (2009) S-RNase triggers mitochondrial alteration and DNA degradation in the incompatible pollen tube of Pyrus pyrifolia in vitro. Plant J 57:220–229PubMedCrossRefGoogle Scholar
  36. Wang CL, Wu J, Xu GH, Gao Y, Chen G, Wu JY, Wu H, Zhang SL (2010) S-RNase disrupts tip-localized reactive oxygen species and induces nuclear DNA degradation in incompatible pollen tubes of Pyrus pyrifolia. J Cell Sci 123:4301–4309PubMedCrossRefGoogle Scholar
  37. Wang L, Lv X, Li H, Zhang M, Wang H, Jin B, Chen T (2013) Inhibition of apoplastic calmodulin impairs calcium homeostasis and cell wall modeling during Cedrus deodara pollen tube growth. PLoS ONE 8:e55411PubMedCentralPubMedCrossRefGoogle Scholar
  38. Wilkins KA, Bancroft J, Bosch M, Ings J, Smirnoff N, Franklin-Tong VE (2011) Reactive oxygen species and nitric oxide mediate actin reorganization and programmed cell death in the self-incompatibility response of Papaver. Plant Physiol 156:404–416PubMedCentralPubMedCrossRefGoogle Scholar
  39. Wu JY, Shang ZL, Wu J, Jiang XT, Moschou PN, Sun WD, Roubelakis-Angelakis KA, Zhang SL (2010) Spermidine oxidase-derived H2O2 regulates pollen plasma membrane hyperpolarization-activated Ca2+-permeable channels and pollen tube growth. Plant J 63:1042–1053PubMedCrossRefGoogle Scholar
  40. Wu JY, Qu HY, Jin C, Shang ZL, Wu J, Xu GH, Gao YB, Zhang SL (2011) cAMP activates hyperpolarization-activated Ca2+ channels in the pollen of Pyrus pyrifolia. Plant Cell Rep 30:1193–1200PubMedCrossRefGoogle Scholar
  41. Yang T, Poovaiah BW (2003) Calcium/calmodulin-mediated signal network in plants. Trends Plant Sci 8:505–512PubMedCrossRefGoogle Scholar
  42. Ye ZH, Sun DY, Guo JF (1988) Preliminary study on wheat cell wall calmodulin. Chin Sci Bull 33:624–626Google Scholar
  43. Zhang SL, Hiratsuka S (1999) Variations in S-protein levels in styles of Japanese pears and the expression of self-incompatibility. J Japan Soc Hort Sci 68:911–918CrossRefGoogle Scholar
  44. Zhang SL, Hiratsuka S (2000) Cultivar and developmental differences in S-protein concentration and self-incompatibility in the Japanese pear. HortScience 35:917–920Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Xueting Jiang
    • 1
  • Yongbin Gao
    • 1
  • Hongsheng Zhou
    • 1
  • Jianqing Chen
    • 1
  • Juyou Wu
    • 1
  • Shaoling Zhang
    • 1
    Email author
  1. 1.College of Horticulture, State Key Laboratory of Crop Genetics and Germplasm EnhancementNanjing Agricultural UniversityNanjingChina

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