胞质ca2参与调节拟南芥下胚轴向光弯曲机制研究-study on the mechanism of cytoplasmic ca2 participating in the regulation of arabidopsis hypocotyls light bending.docx

that the PHOT1 and PHOT2 mediated BL-induced increase in [Ca2+ ]cyt differently, and PHOT2 mainly mediated the BL-induced [Ca2+]cyt signal in A.thaliana etiolated seedlings. By using Ca2+ channel blockers, LaCl3; Ca2+ chelating, EGTA; inhibitors of PLC, U73122 and internal Ca2+ stores inhibitors RR, we demonstrated that PHOT2 mainly induced Ca2+ released to the cytosolic through PLC-dependent signaling. Patch clamp results show that BL can activate PHOT1-dependent plasma membrane Ca2+ channel weaklybut not PHOT2-dependent, which was accordingly with the pharmacological results.BL induced phototropism and inhibited of hypocotyl elongation of etiolated seedlings.Phenotype analysis showed that both PHOT1 and PHOT2 mediated the BL-inhibited hypocotyl elongation under high intensity of BL. Because of the Ca2+ involved in the BL-induced hypocotyl elongation inhibition had beenstudied clearly, here we put our attention to the relationship between BL-induced Ca2+ signal andphototropism. When irradiation with higher fluence rates of BL for 12h, we found that the bend curvature of phot1 was more significant than that in wild-type (gl1) and phot2, with a mean of 75.9±12.6° in phot1 and 51.2±9.8 in gl1. Using [Ca2+]cyt reporter, FRET-Sensitized Emission Imaging of YC3.60, Ca2+differences between illuminated and shaded sides could be found obviously after strong BL-inducedphototropism. All of above indicate that Ca2+ involved in strong BL induced phototropism. In addition, inhibited auxin efflux by NPA strongly suppress the [Ca2+]cyt increase and the phototropism induced by strong BL, especially in phot1 mutant, with a proportion of 82.6% and 77.9%, respectively. So we conclude that the Ca2+ signal was induced by auxin polar transportation. However, yeast-two-hybrid and BiFC results showed that phototropins did not interact with the auxin efflux carrier PIN1/PIN3 in vivo andin vitro, this means PHOT1/ PHOT2 regulate auxin polar transportation by other m