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Calcium signalling mediates self-incompatibility response in the Brassicaceae

Nature Plants volume 1, Article number: 15128 (2015) Cite this article

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An Erratum to this article was published on 03 September 2015

Abstract

Self-incompatibility in the Brassicaceae is controlled by multiple haplotypes encoding the pollen ligand (S-locus protein 11, SP11, also known as S-locus cysteine-rich protein, SCR) and its stigmatic receptor (S-receptor kinase, SRK). A haplotype-specific interaction between SP11/SCR and SRK triggers the self-incompatibility response that leads to self-pollen rejection, but the signalling pathway remains largely unknown. Here we show that Ca2+ influx into stigma papilla cells mediates self-incompatibility signalling. Using self-incompatible Arabidopsis thaliana expressing SP11/SCR and SRK, we found that self-pollination specifically induced an increase in cytoplasmic Ca2+ ([Ca2+]cyt) in papilla cells. Direct application of SP11/SCR to the papilla cell protoplasts induced Ca2+ increase, which was inhibited by D-(−)-2-amino-5-phosphonopentanoic acid (AP-5), a glutamate receptor channel blocker. An artificial increase in [Ca2+]cyt in papilla cells arrested wild-type (WT) pollen hydration. Treatment of papilla cells with AP-5 interfered with self-incompatibility, and Ca2+ increase on the self-incompatibility response was reduced in the glutamate receptor-like channel (GLR) gene mutants. These results suggest that Ca2+ influx mediated by GLR is the essential self-incompatibility response leading to self-pollen rejection.

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Figure 1: Dual pollination assay using self-incompatible A. thaliana.
Figure 2: Dynamics of [Ca2+]cyt in YC3.60pm-expressing papilla cells following self- and cross-pollination.
Figure 3: [Ca2+]cyt dynamics in papilla cell protoplasts treated with SP11/SCR.
Figure 4: A glutamate receptor antagonist AP-5 interferes with self-incompatibility response.
Figure 5: Glutamate receptor-like channel nonsense mutants are compromised in Ca2+ increase on self-incompatibility response.

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Acknowledgements

We thank Y. Ryokume, M. Okamura, F. Yamamoto, and M. Matsumura-Kawashima for their technical assistance. This work was supported in part by Grants-in-Aid for Scientific Research on Innovative Areas (21112003 to M. Iwano; 23113002 to S.T.), Grants-in-Aid for Scientific Research (23570056, 26440145 to M. Iwano; 21248014, 25252021 to S.T.), and Grants-in-Aid for Creative Scientific Research (16GS0316 to A.I.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT); and by Grants-in-Aid for JSPS Research Fellow (12J08273 to K.I.) from the Japan Society for the Promotion of Science (JSPS).

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Author notes

  1. Megumi Iwano & Tetsuyuki Entani

    Present address: †Present address: The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Japan,

  2. Kanae Ito

    Present address: •Present address: Medical Institute of Bioreguration, Kyushu University, Fukuoka, Japan,

  3. Mitsuru Kakita

    Present address: ‣Present address: Graduate School of Science, Nagoya University, Chikusa, Japan,

  4. Hiroshi Shiba

    Present address: ․Present address: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan,

  5. Megumi Iwano, Kanae Ito and Sota Fujii: These authors contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0192, Japan

    Megumi Iwano, Kanae Ito, Sota Fujii, Mitsuru Kakita, Hiroko Asano-Shimosato, Motoko Igarashi, Pulla Kaothien-Nakayama, Tetsuyuki Entani, Asaka Kanatani, Masashi Takehisa, Masaki Tanaka, Kunihiko Komatsu, Hiroshi Shiba, Akira Isogai & Seiji Takayama

  2. The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, 567-0047, Osaka, Japan

    Takeharu Nagai

  3. Brain Science Institute, The Institute of Physical and Chemical Research (RIKEN), Wako, 351-0198, Saitama, Japan

    Atsushi Miyawaki

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Contributions

M. Iwano, A.I. and S.T. initiated the project. M. Iwano, K.I., M.K., P.K.-N., T.N., A.M. and S.T. designed experiments. M. Iwano, K.I., H.A.-S., S.F., M. Igarashi, T.E., A.K., M.T., M.T., K.K. and H.S. performed the experiments. M. Iwano, K.I., S.F. and S.T. wrote the manuscript.

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Correspondence to Seiji Takayama.

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Iwano, M., Ito, K., Fujii, S. et al. Calcium signalling mediates self-incompatibility response in the Brassicaceae. Nature Plants 1, 15128 (2015). https://doi.org/10.1038/nplants.2015.128

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