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 BA 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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