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Identifcation and expression of  DoCCaMK during Sebacina sp.  symbiosis of Dendrobium ofcinale

Yong-MeiXing1,4, Ming-MingZhao1,2,4, Li-ChengGuo2,3, Bing Li1, Juan Chen1 ✉ & ShunXingGuo1 ✉

Dendrobium ofcinale Kimura et Migo is a famous precious medicinal plant in China. Seed and seedling  were c*ted with the mycorrhizal fungus Sebacina sp. CCaMK was initially cloned from D. ofcinale based on a SSH cDNA library of symbiotically germinated seeds with Sebacina sp. Phylogenetic  analysis was performed among DoCCaMK and other CCaMKs. The particle bombardment technique  was used to visualize DoCCaMK-GFP. qRT-PCR and western blot analysis were conducted to determine  the tissue expression patterns of DoCCaMK with (SGS) and without (UGS) Sebacina sp. Furthermore,  the efect of KN-93 on CCaMK expression was also examined. Using NMT the net Ca2+ fuxes and the  CCaMK concentration were measured during D. ofcinale seed germination. DoCCaMK had the highest  homology with Lilium longiforum CCaMK. The DoCCaMK-GFP protein localized in the nucleus and cell  membrane. CCaMK expression was signifcantly upregulated after symbiosis with Sebacina sp. KN-93  could be used as an inhibitor of CCaMK to inhibit D. ofcinale seed germination. Ca2+ infux and the  concentration of the CCaMK in the SGS group was signifcantly more than that of the UGS group. The  characterization of CCaMK provides certain genetic evidence for the involvement of this gene during  seed germination and mycorrhizal c*tion in D. ofcinale.