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Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Characterisation and immunolocalisation of a pollen-specific calmodulin-binding protein from rice

Qiusheng Zhang A , Jingjing Zhang A , Daichang Yang A , Yangsheng Li A , Shaoqing Li A and Yingguo Zhu A B
+ Author Affiliations
- Author Affiliations

A Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, People’s Republic of China.

B Corresponding author. Email: zhuyg@public.wh.hb.cn

Functional Plant Biology 33(6) 555-562 https://doi.org/10.1071/FP05320
Submitted: 23 December 2005  Accepted: 10 March 2006   Published: 1 June 2006

Abstract

A novel calmodulin (CaM)-binding protein of rice (Oryza sativa pollen CaM-binding protein, OsPCBP) was isolated. It contains six tetratrcopeptide repeats (TPRs) with a molecular mass of ~76.7 kDa. Database searches show that OsPCBP is conserved in monocots and dicots. The results from CaM–agarose pull-down assays show that OsPCBP binds to CaM in a Ca2+-dependent manner and its CaM-binding domain (CBD) is located in a segment VSKGWRLLALVLSAQQRY. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis have demonstrated that OsPCBP is a pollen-specific and late-expressed gene. Immunolocalisation showed that OsPCBP was localised in the amyloplast and intine during the late stage of pollen development. However, upon pollen hydration, there was some leakage of protein and the distribution of OsPCBP in the intine varied with the duration of hydration. After pollen germination, much OsPCBP was observed in the pollen tube wall. These results indicate the relationship of OsPCBP with starch accumulation and its involvement in pollen germination.

Keywords: amyloplast, calmodulin-binding protein, immunolocalisation, intine, Oryza sativa L., pollen.


Acknowledgments

We are very grateful to Binglian Xu (Nanjing Agricultural University, China) and Prof. Jie Zhao (Wuhan University, China) for critical reading of the manuscript. We thank Jing Yang and Yuan Qin for technical assistance. This work was supported by the Chinese National ‘973’ Grant Program (No. 2001CB108806).


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