Calreticulin expression in relation to exchangeable Ca2+ level that changes dynamically during anthesis, progamic phase, and double fertilization in Petunia

Abstract

Calcium (Ca2+) plays essential roles in plant sexual reproduction, but the sites and the mechanism of Ca2+ mobile storage during pollen–pistil interactions have not been fully defined. Because the Ca2+-buffering protein calreticulin (CRT) is able to bind and sequester Ca2+, it can serve as a mobile intracellular store of easily releasable Ca2+ and control its local concentration within the cytoplasm. Our previous studies showed an enhanced expression of Petunia hybrida CRT gene (PhCRT) during pistil transmitting tract maturation, pollen germination and tube outgrowth on the stigma, gamete fusion, and early embryogenesis. Here, we demonstrate that elevated expression of CRT results in the accumulation of this protein in response to anthesis, pollination, sperm cells deposition within the receptive synergid and fertilization, when the level of exchangeable Ca2+ changes dynamically. CRT localizes mainly to the endoplasmic reticulum and Golgi compartments in the pistil transmitting tract cells, germinated pollen/tubes, and sporophytic/gametophytic cells of the ovule and corresponds with loosely bound Ca2+. Additionally, the immunogold research shows, for the first time, highly selective CRT distribution in specific nuclear sub-domains. On the basis of our results, we discuss the possible functions of CRT with respect to the critical role of Ca2+ homeostasis during key events of the multi-step process of generative reproduction in angiosperms.