title: Spermidine oxidase-derived H₂O₂ regulates pollen plasma membrane hyperpolarization-activated Ca(2+) -permeable channels and pollen tube growth.
abstract: Spermidine (Spd) has been correlated with various physiological and developmental processes in plants, including pollen tube growth. In this work, we show that Spd induces an increase in the cytosolic Ca(2+) concentration that accompanies pollen tube growth. Using the whole-cell patch clamp and outside-out single-channel patch clamp configurations, we show that exogenous Spd induces a hyperpolarization-activated Ca(2+) current: the addition of Spd cannot induce the channel open probability increase in excised outside-out patches, indicating that the effect of Spd in the induction of Ca(2+) currents is exerted via a second messenger. This messenger is hydrogen peroxide (H₂O₂), and is generated during Spd oxidation, a reaction mediated by polyamine oxidase (PAO). These reactive oxygen species trigger the opening of the hyperpolarization-activated Ca(2+) -permeable channels in pollen. To provide further evidence that PAO is in fact responsible for the effect of Spd on the Ca(2+) -permeable channels, two Arabidopsis mutants lacking expression of the peroxisomal-encoding AtPAO3 gene, were isolated and characterized. Pollen from these mutants was unable to induce the opening of the Ca(2+) -permeable channels in the presence of Spd, resulting in reduced pollen tube growth and seed number. However, a high Spd concentration triggers a Ca(2+) influx beyond the optimal, which has a deleterious effect. These findings strongly suggest that the Spd-derived H₂O₂ signals Ca(2+) influx, thereby regulating pollen tube growth.
authors.affiliation: College of Horticulture, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Juyou
authors.initials: J
authors.lastname: Wu
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Shang
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.forename: Xueting
authors.initials: X
authors.lastname: Jiang
authors.forename: Panagiotis N
authors.initials: PN
authors.lastname: Moschou
authors.forename: Wending
authors.initials: W
authors.lastname: Sun
authors.forename: Kalliopi A
authors.initials: KA
authors.lastname: Roubelakis-Angelakis
authors.forename: Shaoling
authors.initials: S
authors.lastname: Zhang
mesh_terms: D015220:Calcium Channels / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D008856:Microscopy, Fluorescence; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000235:genetics / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011058:Pollen / Q000378:metabolism*; D053205:Pollen Tube / Q000254:growth & development / Q000378:metabolism*; D015221:Potassium Channels / Q000378:metabolism*; D013095:Spermidine / Q000378:metabolism; D000097667:Polyamine Oxidase
chemical_list: D015220:Calcium Channels; D054815:Cyclic Nucleotide-Gated Cation Channels; D006861:Hydrogen Peroxide; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D000587:Oxidoreductases Acting on CH-NH Group Donors; D015221:Potassium Channels; D013095:Spermidine; D000097667:Polyamine Oxidase