Abstract
The pollen receptor kinases (PRK) are critical regulators of pollen tube growth. The Arabidopsis genome encodes eight PRK genes, of which six are highly expressed in pollen tubes. The potential functions of AtPRK1 through AtPRK5, but not of AtPRK6, in pollen growth were analyzed in tobacco. Herein, AtPRK6 was cloned, and its function was identified. AtPRK6 was expressed specifically in pollen tubes. A yeast two-hybrid screen of AtPRK6 against 14 Arabidopsis Rop guanine nucleotide exchange factors (RopGEFs) showed that AtPRK6 interacted with AtRopGEF8 and AtRopGEF12. These interactions were confirmed in Arabidopsis mesophyll protoplasts. The interactions between AtPRK6 and AtRopGEF8/12 were mediated by the C-termini of AtRopGEF8/12 and by the juxtamembrane and kinase domain of AtPRK6, but were not dependent on the kinase activity. In addition, transient overexpression of AtPRK6::GFP in Arabidopsis protoplasts revealed that AtPRK6 was localized to the plasma membrane. Tobacco pollen tubes overexpressing AtPRK6 exhibited shorter tubes with enlarged tips. This depolarized tube growth required the kinase domain of AtPRK6 and was not dependent on kinase activity. Taken together, the results show that AtPRK6, through its juxtamembrane and kinase domains (KD), interacts with AtRopGEF8/12 and plays crucial roles in polarized growth of pollen tubes.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alvarez-Buylla, E.R., Benítez, M., Corverapoiré, A., Álvaro Chaos Cador, Folter, S.D., Buen, A. G.D., Garay-Arroyo, A., García-Ponce, B. Fabiola Jaimes-Miranda, F., Pérez-Ruiz, R.V., Piñeyro-Nelson, A., and Sánchez-Corrales, Y.E. (2015). Flower development. In: Arabidopsis Book. (Passaic: Humana Press), p. e0127.
Arthur, K.M., Vejlupkova, Z., Meeley, R.B., Fowler, J.E. (2004). Maize ROP2 GTPase provides a competitive advantage in male gametophyte. Genetics 165, 2137–2151.
Bassham, D.C., and Raikhel, N.V. (2000). Unique features of the plant vacuolar sorting machinery. Curr Opin Cell Biol 12, 491–495.
Berken, A., Thomas, C., and Wittinghofer, A. (2005). A new family of RhoGEFs activates the Rop molecular switch in plants. Nature 436, 1176–1180.
Chang, F., Gu, Y., Ma, H., and Yang, Z. (2013). AtPRK2 promotes ROP1 activation via RopGEFs in the control of polarized pollen tube growth. Mol Plant 6, 1187–1201.
Chou, C.K., Dull, T.J., Russell, D.S., Ullrich, A. and Rosen, O.M. (1987) Human insulin receptors mutated at the ATP-binding site lack protein tyrosine kinase activity and fail to mediate postreceptor effects of insulin. J Biol Chem 262, 1842–1847.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method forAgrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735–743.
Fu, Y. (2015). The cytoskeleton in the pollen tube. Curr Opin Plant Biol 28, 111–119.
Fu, Y., Wu, G., and Yang, Z. (2001). RopGTPase-dependent dynamics of tip-localized F-actin controls tip growth in pollen tubes. J Cell Biol 152, 1019–1032.
Gish, L.A., and Clark, S.E. (2011). The RLK/Pelle family of kinases. Plant J 66, 117–127.
Gu, Y., Li, S., Lord, E.M., and Yang, Z. (2006). Members of a novel class of Arabidopsis Rho guanine nucleotide exchange factors control Rho GTPase-dependent polar growth. Plant Cell 18, 366–381.
Gu, Y., Wang, Z., and Yang, Z. (2004). ROP/RAC GTPase: an old new master regulator for plant signaling. Curr Opin Plant Biol 7, 527–536.
Gu, Y., Fu, Y., Dowd, P., Li, S., Vernoud, V., Gilroy, S., and Yang, Z. (2005). A Rho family GTPase controls actin dynamics and tip growth via two counteracting downstream pathways in pollen tubes. J Cell Biol 169, 127–138.
Gu, Y., Vernoud, V., Fu, Y., and Yang, Z. (2003). ROP GTPase regulation of pollen tube growth through the dynamics of tip-localized F-actin. J Exp Bot 54, 93–101.
Guan, Y., Guo, J., Li, H., and Yang, Z. (2013). Signaling in pollen tube growth: crosstalk, feedback, and missing links. Mol Plant 6, 1053–1064.
Gui, C.P., Dong, X., Liu, H.K., Huang, W.J., Zhang, D., Wang, S.J., Barberini, M.L., Gao, X.Y., Muschietti, J., McCormick, S., and Tang, W.H. (2014). Overexpression of the tomato pollen receptor kinase LePRK1 rewires pollen tube growth to a blebbing mode. Plant Cell 26, 3538–3555.
Hwang, J.U., Gu, Y., Lee, Y.J., and Yang, Z. (2005). Oscillatory ROP GTPase activation leads the oscillatory polarized growth of pollen tubes. Mol Biol Cell 16, 5385–5399.
Jefferson, R.A., Kavanagh, T.A., and Bevan, M.W. (1987). GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6, 3901–3907.
Johnson, M.A., and Preuss, D. (2003). On your mark, get set, GROW! LePRK2-LAT52 interactions regulate pollen tube growth. Trends Plant Sci 8, 97–99.
Kaothien, P., Ok, S.H., Shuai, B., Wengier, D., Cotter, R., Kelley, D., Kiriakopolos, S., Muschietti, J., and McCormick, S. (2005). Kinase partner protein interacts with the LePRK1 and LePRK2 receptor kinases and plays a role in polarized pollen tube growth. Plant J 42, 492–503.
Khamsuk, O. (2011). Functions of leucine rich repeat receptor-like kinases (LRR-RLKs), in Arabidopsis sexual reproduction. Doctoral Dissertation. (California: University of California Riverside).
Kim, H.U., Cotter, R., Johnson, S., Senda, M., Dodds, P., Kulikauskas, R., Tang, W., Ezcurra, I., Herzmark, P., and McCormick, S. (2002). New pollen-specific receptor kinases identified in tomato, maize and Arabidopsis: the tomato kinases show overlapping but distinct localization patterns on pollen tubes. Plant Mol Biol 50, 1–16.
Lee, H.S., Karunanandaa, B., McCubbin, A., Gilroy, S., and Kao, T. (1996). PRK1, a receptor-like kinase of Petunia inflata, is essential for postmeiotic development of pollen. Plant J 9, 613–624.
Lee, Y.J., Szumlanski, A., Nielsen, E., and Yang, Z. (2008). Rho-GTPase–dependent filamentous actin dynamics coordinate vesicle targeting and exocytosis during tip growth. J Cell Biol 181, 1155–1168.
Lennon, K.A., and Lord, E.M. (2000). In vivo pollen tube cell of Arabidopsis thaliana. I. Tube cell cytoplasm and wall. Protoplasma 214, 45–56.
Lew, R.R. (2004). Osmotic effects on the electrical properties of Arabidopsis root hair vacuoles in situ. Plant Physiol 134, 352–360.
Li, H., Lin, Y., Heath, R.M., Zhu, M.X., and Yang, Z.B. (1999). Control of pollen tube tip growth by a Rop GTPase-dependent pathway that leads to tip-localized calcium influx. Plant Cell 11, 1731–1742.
Li, H., and Yang, W.C. (2016). RLKs orchestrate the signaling in plant malefemale interaction. Sci China Life Sci 59, 867–877.
Li, L., Kim, B.G., Cheong, Y.H., Pandey, G.K., and Luan, S. (2006). A Ca2 signaling pathway regulates a K2 channel for low-K response in Arabidopsis. Proc Natl Acad Sci USA 103, 12625–12630.
Li, Z., and Liu, D. (2012). ROPGEF1 and ROPGEF4 are functional regulators of ROP11 GTPase in ABA-mediated stomatal closure in Arabidopsis. FEBS Lett 586, 1253–1258.
Lin, Y., Li, Y., Zhu, Y., Zhang, J., Li, Y., Liu, X., Jiang, W., Yu, S., You, X.F., Xiao, C., Hong, B., Wang, Y., Jiang, J.D., and Si, S. (2012). Identification of antituberculosis agents that target ribosomal protein interactions using a yeast two-hybrid system. Proc Natl Acad Sci USA 109, 17412–17417.
MacRobbie, E.A. (2006). Osmotic effects on vacuolar ion release in guard cells. Proc Natl Acad Sci USA 103, 1135–1140.
Mu, J.H., Lee, H.S., and Kao, T.H. (1994). Characterization of a pollenexpressed receptor-like kinase gene of Petunia inflata and the activity of its encoded kinase. Plant Cell 6, 709–721.
Muschietti, J., Eyal, Y., and Mccormick, S. (1998) Pollen tube localization implies a role in pollen-pistil interactions for the tomato receptor-like protein kinases LePRK1 and LePRK2. Plant Cell 10, 319–330.
Qin, Y., and Yang, Z. (2011). Rapid tip growth: insights from pollen tubes. Semin Cell Dev Biol 22, 816–824.
Salem, T., Mazzella, A., Barberini, M.L., Wengier, D., Motillo, V., Parisi, G., and Muschietti, J. (2011). Mutations in two putative phosphorylation motifs in the tomato pollen receptor kinase LePRK2 show antagonistic effects on pollen tube length. J Biol Chem 286, 4882–4891.
Tang, W., Ezcurra, I., Muschietti, J., and McCormick, S. (2002). A cysteinerich extracellular protein, LAT52, interacts with the extracellular domain of the pollen receptor kinase LePRK2. Plant Cell 14, 2277–2287.
Tang, W., Kelley, D., Ezcurra, I., Cotter, R., and McCormick, S. (2004). LeSTIG1, an extracellular binding partner for the pollen receptor kinases LePRK1 and LePRK2, promotes pollen tube growth in vitro. Plant J 39, 343–353.
Takeuchi, H., and Higashiyama, T. (2016). Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis. Nature 531, 245–248.
Thomas, C., and Berken, A. (2010). Structure and function of ROPs and their GEFs. (Berlin Heidelberg: Springer), pp. 49–69.
Twell, D., Yamaguchi, J., Wing, R.A., Ushiba, J., and McCormick, S. (1991). Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev 5, 496–507.
Twell, D., Klein, T.M., Fromm, M.E., and McCormick, S. (1989). Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiol 91, 1270–1274.
Vaid, N., Macovei, A., and Tuteja, N. (2013). Knights in action: lectin receptor-like kinases in plant development and stress responses. Mol Plant 6, 1405–1418.
Wang, T., Liang, L., Xue, Y., Jia, P.F., Chen, W., Zhang, M.X., Wang, Y.C., Li, H.J., and Yang, W.C. (2016). A receptor heteromer mediates the male perception of female attractants in plants. Nature 531, 241–244.
Yang, T., Wang, L., Li, C., Liu, Y., Zhu, S., Qi, Y., Liu, X., Lin, Q., Luan, S., and Yu, F. (2015). Receptor protein kinase FERONIA controls leaf starch accumulation by interacting with glyceraldehyde-3-phosphate dehydrogenase. Biochem Biophys Res Commun 465, 77–82.
Yang, Z. (2008). Cell polarity signaling in Arabidopsis. Annu Rev Cell Dev Biol 24, 551–575.
Yang, Z., and Fu, Y. (2007). ROP/RAC GTPase signaling. Curr Opin Plant Biol 10, 490–494.
Yoo, S.D., Cho, Y.H., and Sheen, J. (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2, 1565–1572.
Zhang, Y., and McCormick, S. (2008). Regulation of pollen tube polarity. Plant Signal Behav 3, 345–347.
Zhang, D., Wengier, D., Shuai, B., Gui, C.P., Muschietti, J., McCormick, S., and Tang, W.H. (2008). The pollen receptor kinase LePRK2 mediates growth-promoting signals and positively regulates pollen germination and tube growth. Plant Physiol 148, 1368–1379.
Zhang, Y., and McCormick, S. (2007). A distinct mechanism regulating a pollen-specific guanine nucleotide exchange factor for the small GTPase Rop in Arabidopsis thaliana. Proc Natl Acad Sci USA 104, 18830–18835.
Zhao, X.Y., Wang, Q., Li, S., Ge, F.R., Zhou, L.Z., McCormick, S., and Zhang, Y. (2013). The juxtamembrane and carboxy-terminal domains of Arabidopsis PRK2 are critical for ROP-induced growth in pollen tubes. J Exp Bot 64, 5599–5610.
Zhou, J.M., and Yang, W.C. (2016). Receptor-like kinases take center stage in plant biology. Sci China Life Sci 59, 863–866.
Zhou, L.M., Lan, W.Z., Jiang, Y.Q., Fang, W., and Luan, S. (2014). A calcium-dependent protein kinase interacts with and activates acalcium channel to regulate pollen tube growth. Mol Plant 7, 7361–7372.
Zhu, L., Zhang, Y., Kang, E., Xu, Q., Wang, M., Rui, Y., Liu, B., Yuan, M., and Fu, Y. (2013). MAP18 regulates the direction of pollen tube growth in Arabidopsis by modulating F-actin organization. Plant Cell 25, 851–867.
Zou, Y., Aggarwal, M., Zheng, W.G., Wu, H.M., and Cheung, A.Y. (2011). Receptor-like kinases as surface regulators for RAC/ROP-mediated pollen tube growth and interaction with the pistil. AoB Plants 2011, plr017.
Acknowledgements
The authors are grateful to Dr. Jing Li for reading and giving comments to this manuscript. This work was supported by the National Natural Science Foundation of China (31300247).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Figure S1
Large vacuoles accumulated at the tips of pollen tubes overexpressing AtPRK6.
Figure S2 Pollengermination rate of prk6 mutants. Pictures were taken after 5-h incubation on pollen germination medium (left). Percentage of germinated WT and prk6 mutants (right).
Figure S3 AtRopGEF8 induced pollen tube depolarization when expressed in tobacco pollen tubes.
Rights and permissions
About this article
Cite this article
Yu, Y., Song, J., Tian, X. et al. Arabidopsis PRK6 interacts specifically with AtRopGEF8/12 and induces depolarized growth of pollen tubes when overexpressed. Sci. China Life Sci. 61, 100–112 (2018). https://doi.org/10.1007/s11427-016-9107-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-016-9107-3