P2K1 Receptor, Heterotrimeric Gα Protein and CNGC2/4 Are Involved in Extracellular ATP-Promoted Ion Influx in the Pollen of Arabidopsis thaliana.

As an apoplastic signal, extracellular ATP (eATP) is involved in plant growth and development. eATP promotes tobacco pollen germination (PG) and pollen tube growth (PTG) by stimulating Ca2+ or K+ absorption. Nevertheless, the mechanisms underlying eATP-stimulated ion uptake and their role in PG and PTG are still unclear. Here, ATP addition was found to modulate PG and PTG in 34 plant species and showed a promoting effect in most of these species. Furthermore, by using Arabidopsis thaliana as a model, the role of several signaling components involved in eATP-promoted ion (Ca2+, K+) uptake, PG, and PTG were investigated. ATP stimulated while apyrase inhibited PG and PTG. Patch-clamping results showed that ATP promoted K+ and Ca2+ influx into pollen protoplasts. In loss-of-function mutants of P2K1 (dorn1-1 and dorn1-3), heterotrimeric G protein α subunit (gpa1-1, gpa1-2), or cyclic nucleotide gated ion channel (cngc2, cngc4), eATP-stimulated PG, PTG, and ion influx were all impaired. Our results suggest that these signaling components may be involved in eATP-promoted PG and PTG by regulating Ca2+ or K+ influx in Arabidopsis pollen grains.

Extracellular ATP promoted pollen germination and tube growth of Nicotiana tabacum through promoting K+ and Ca2+ absorption.

Extracellular ATP (eATP) plays an essential role in plant growth, development, and stress tolerance. Here, we report that eATP participated in Nicotiana tabacum pollen germination (PG) and pollen tube growth (PTG) by regulating K+ and Ca2+ influx. Exogenous ATP or ADP effectively promoted PG and PTG in a dose-dependent manner; weakly hydrolysable ATP analog (ATPγS) showed a similar effect. AMP, adenosine, adenine, and phosphate did not affect PG or PTG. Within a certain range, higher concentrations of K+ or Ca2+ in the medium increased the effect of ATP in promoting PG and PTG. However, in mediums containing K+ or Ca2+ concentrations above this range, the effect of ATP was reversed, resulting in PG and PTG inhibition. Ca2+ chelators (EGTA), Ca2+ channel blockers, and K+ channel blockers suppressed ATP-promoted PG and PTG. Results from a patch clamp showed that ATP activated a K+ and Ca2+ influx in pollen protoplasts. These results suggest that, as an apoplastic signal, eATP may be involved in PG and PTG via regulating Ca2+ and K+ absorption.