Abstract
BackgroundThe functional characteristics of SLAC/SLAH family members isolated from Arabidopsis thaliana, poplar, barley and rice have been comprehensively investigated. However, there are no reports regarding SLAC/SLAH family genes from Rosaceae plants.ResultsIn this study, the function of PbrSLAH3, which is predominately expressed in pear (Pyrus bretschneideri) root, was investigated. PbrSLAH3 can rescue the ammonium toxicity phenomenon of slah3 mutant plants under high-ammonium/low-nitrate conditions. In addition, yeast two-hybrid and bimolecular fluorescence complementation assays confirmed that PbrSLAH3 interacts with PbrCPK32. Moreover, when PbrSLAH3 was co-expressed with either the Arabidopsis calcium-dependent protein kinase (CPK) 21 or PbrCPK32 in Xenopus oocytes, yellow fluorescence was emitted from the oocytes and typical anion currents were recorded in the presence of extracellular NO3−. However, when PbrSLAH3 alone was injected, no yellow fluorescence or anion currents were recorded, suggesting that anion channel PbrSLAH3 activity was controlled through phosphorylation. Finally, electrophysiological and transgene results showed that PbrSLAH3 was more permeable to NO3− than Cl−.ConclusionWe suggest that PbrSLAH3 crossing-talk with PbrCPK32 probably participate in transporting of nitrate nutrition in pear root.
Highlights
The functional characteristics of SLAC/SLAH family members isolated from Arabidopsis thaliana, poplar, barley and rice have been comprehensively investigated
PbrSLAC1 was exclusively expressed in leaf, while PbrSLAH3, PbrSLAH2/3–2 and PbrSLAH2/3–3 were widely expressed in root, leaf and pollen
The transcriptome data analysis suggested that the expression level of PbrSLAH3 was up-regulated during nitrate starvation and after the re-supply of nitrate, its expression level was gradually down-regulated (Additional file 1: Figure S1).Taken together these results, we presume that PbrSLAH3 may be involved in nitrate transport in pear root and may play an important role in nitrate nutrition in pear trees
Summary
The functional characteristics of SLAC/SLAH family members isolated from Arabidopsis thaliana, poplar, barley and rice have been comprehensively investigated. SLAC/SLAH family genes (S-type anion channels) from species, including Arabidopsis thaliana, poplar, barley and rice, have been identified and studied, five members in Arabidopsis [8, 9], seven members in Populus [10] and nine members in rice [11]. AtSLAC1, together with SLAC1 homologous 3 (AtSLAH3), plays an important role in regulating ABA-induced stomatal closure [16,17,18,19] During this process, ABA is firstly perceived by the RCAR/PYR/ PYL–ABI1 complex, which releases protein kinase from ABI1-induced inhibition, and AtSLAC1 and AtSLAH3 are activated by the calcium-independent protein kinase OST1 and the calcium-dependent protein kinases (CPKs) of Arabidopsis [13]. The results from the Geiger group showed that SLAH1, as a silent subunit, facilitates SLAH3-mediated chloride efflux from pericycle
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