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

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.

Structural cell wall proteins from five pollen species and their relationship with boron

Boron (B) is an essential micronutrient for the survival of vascular plants. The most severe effect derived from a deficiency of B is the alteration of cell wall biogenesis and pollen germination. We investigated pollen of plant species that require B for germination (Zea may L. and Nicotiana tabacum L.), as well as those that can germinate without B (Pinus sp, Lilum longiflorum, Impatiens sp.). In both groups, B addition in the growth medium increased the length of the pollen tube after germination. Hydroxyproline Rich Glycoproteins (HRGPs) are the most abundant cell wall structural proteins of dicotyledonous plants and the sexual tissues of monocotyledonous plants. Here, we show that maize pollen accumulated a significant pool of hydroxyproline (Hyp) and 63% of this amino acid was localized in the pollen tube wall. Maize pollen germinated in the presence of B accumulated soluble (48%) and non-soluble (16%) Hyp in the pollen tube wall in contrast to maize pollen germinated without B. In addition, B seems to modify the amount of HRGPs that become cross-linked to the wall. Immunolocalization of HRGPs showed that these glycoproteins were preferentially localized in the pollen tube of maize, not in the pollen grain itself. Hence, B might affect the assembling of HRGPs in the wall of pollen tubes grown in vitro.