PbrSLAH3 is a nitrate-selective anion channel which is modulated by calcium-dependent protein kinase 32 in pear
PbrSLAH3 is a nitrate-selective anion channel which is modulated by calcium-dependent protein kinase 32 in pear
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DOI：
10.1186/s12870-019-1813-z
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发表时间：
2019-05-08
影响因子：
5.3
通讯作者：
Wu, Juyou
中科院分区：
文献类型：
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作者：
Chen, Guodong;Wang, Li;Wu, Juyou
研究背景对从拟南芥、杨树、大麦和水稻中分离的SLAC/SLAH家族成员的功能特性进行了全面的研究。然而，目前还没有关于蔷薇科植物SLAC/SLAH家族基因的报道。结果在本研究中，研究了主要在梨(Pyrus bretschneideri)根中表达的PbrSLAH3的功能。 PbrSLAH3可以挽救slah3突变体植物在高铵/低硝条件下的铵毒性现象。此外，酵母双杂交和双分子荧光互补测定证实PbrSLAH3与PbrCPK32相互作用。此外，当 PbrSLAH3 与拟南芥钙依赖性蛋白激酶 (CPK) 21 或 PbrCPK32 在非洲爪蟾卵母细胞中共表达时，卵母细胞会发出黄色荧光，并且在细胞外 NO3- 存在的情况下记录到典型的阴离子电流。然而，当单独注射 PbrSLAH3 时，没有记录到黄色荧光或阴离子电流，表明阴离子通道 PbrSLAH3 活性是通过磷酸化控制的。最后，电生理和转基因结果表明PbrSLAH3对NO3-的渗透性比Cl-更强。结论我们认为PbrSLAH3与PbrCPK32的相互作用可能参与了梨根中硝酸盐营养的转运。
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.