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Published on March 2, 2007; 10.1104/pp.106.095273

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Received December 26, 2006
Accepted February 17, 2007

Actin Dynamics in Papilla Cells of Brassica rapa during Self- and Cross-pollination

Megumi Iwano *, Hiroshi Shiba , Kyoko Matoba , Teruhiko Miwa , Miyuki Funato , Tetsuyuki Entani , Pulla Nakayama , Hiroko Shimosato , Akio Takaoka , Akira Isogai , and Seiji Takayama

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0101, Japan; Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, Suita, Osaka 565-0781, Japan

* Corresponding author; email: m-iwano{at}bs.naist.jp.

The self-incompatibility (SI) system of the plant species Brassica is controlled by the S-locus, which contains SRK (S-receptor kinase) and SP11 (S-locus protein 11). SP11 binding to SRK induces SRK autophosphorylation and initiates a signaling cascade leading to the rejection of self-pollen. However, the mechanism controlling hydration and germination arrest during self-pollination is unclear. In this study, we examined the role of actin, a key cytoskeletal component regulating the transport system for hydration and germination, in the papilla cell during pollination. Using rhodamine-phalloidin staining, we showed that cross-pollination induced actin polymerization, while self-pollination induced actin reorganization and likely depolymerization. By monitoring transiently expressed GFP-mTalin, we observed the concentration of actin bundles at the cross-pollen attachment site and actin reorganization and likely depolymerization at self-pollen attachment site, the results corresponding to those obtained by rhodamine-phalloidin staining. We further showed that the coat of self-pollen is sufficient to mediate this response. The actin-depolymerizing drug cytochalasin D (CD) significantly inhibited pollen hydration and germination during cross-pollination, further emphasizing a role for actin in these processes. Additionally, three-dimensional electron microscopic tomography revealed the close association of the actin cytoskeleton with an apical vacuole network. Self-pollination disrupted the vacuole network while cross-pollination led to vacuolar rearrangements toward the site of pollen attachment. Taken together, our data suggest that self- and cross-pollination differentially affect the dynamics of actin cytoskeleton leading to changes in vacuolar structure associated with hydration and germination.




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