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Plant and Cell Physiology Advance Access published online on July 4, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp100
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Pollen-specific SKP1-like proteins are components of functional SCF complexes and essential for lily pollen tube elongation

Liang-Chi Chang1,2, Cian-Ling Guo2, Yun-Shan Lin2, Hongyong Fu2, Co-Shing Wang3 and Guang-Yuh Jauh1,2,*

1 Institute of Life Science, National Defense Medical Center, Taipei, Taiwan, ROC.
2 Institute of Plant and Microbial Biology, Academia Sinica, Taipei 115, Taiwan, ROC.
3 Graduate Institute of Biotechnology, National Chung-Hsing University, Taichung 402, Taiwan, ROC.

*Corresponding author: Dr. Guang-Yuh Jauh, Institute of Plant and Microbial Biology, Academia Sinica Nankang, Taipei 11529, Taiwan, R.O.C. Phone: 886-2-27871156, Fax: 886-2-27827954, E-mail: jauh{at}gate.sinica.edu.tw


  Abstract

The ubiquitin-proteasome pathway mediates protein degradation and is involved in diverse plant development and differentiation aspects, including pollen tube elongation and self-incompatibility. We characterized three lily (Lilium longiflorum) SKP1-like genes, LSK1-3, that are specifically expressed in late pollen developmental stages and the elongating pollen tube. Encoded peptide sequences reveal that LSK1-3 share high identity to Arabidopsis ASK1 and contain putative N-terminal CUL1 and C-terminal F-box interacting domains. Yeast two-hybrid and in vitro affinity binding assays revealed that the LSKs associate with lily CULLIN1. In addition, the LSKs can functionally complement the yeast skp1 deletion mutant YDR328C. To investigate their biological functions in pollen tube elongation, an in vivo approach for GFP-tagged dominant-negative LSK1-3 was developed. Microprojectile bombardment with N-terminal-truncated LSK1-3 (LSK1-3{Delta}-GFP) significantly retarded pollen tube elongation on both in vitro germination and in vivo self- and cross-pollination after more than 12-hr incubation. Interestingly, elongation of pollen tubes harboring overexpressed LSK2{Delta}-GFP and LSK3{Delta}-GFP was substantially inhibited within the self-pollinated styles. The elongation of most LSK2{Delta}-GFP-transformed pollen tubes could germinate only on the stigmatic surface and showed statistically significant growth arrest as compared with control pollen tubes. Lily exhibits typical gametophytic self-incompatibility via an unknown mechanism, but LSK2 and LSK3 may be involved in this complex machinery. These results suggest critical roles for LSK1-3 in regulating fundamental pollen tube elongation in vitro and in vivo and the 26S proteasome-mediated protein pathway playing an important role in pollen tube elongation.

Keywords: lily (Lilium longiflorum) - LLCUL1 - LSK - pollen tube elongation - SCF complex - self-incompatibility

(Received June 2, 2009; Accepted July 1, 2009)
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