异位表达葡萄硝酸盐转运蛋白 VvNPF6.5 提高拟南芥中的硝酸盐含量和氮利用效率。
Ectopic expression of a grape nitrate transporter VvNPF6.5 improves nitrate content and nitrogen use efficiency in Arabidopsis.
机构信息
Research Institute of Forestry and Pomology, Shanghai Key Lab of Protected Horticultural Technology, Shanghai Academy of Agricultural Sciences, Shanghai, China.
Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
出版信息
BMC Plant Biol. 2020 Dec 7;20(1):549. doi: 10.1186/s12870-020-02766-w.
BACKGROUND
Nitrate plays an important role in grapevines vegetative and reproductive development. However, how grapevines uptake, translocate and utilize nitrate and the molecular mechanism still remains to be investigated.
RESULTS
In this study, we report the functional characterization of VvNPF6.5, a member of nitrate transporter 1/peptide transporter family (NRT1/PTR/NPF) in Vitis vinifera. Subcellular localization in Arabidopsis protoplasts indicated that VvNPF6.5 is plasma membrane localized. Quantitative RT-PCR analysis indicated that VvNPF6.5 is expressed predominantly in roots and stems and its expression is rapidly induced by nitrate. Functional characterization using cRNA-injected Xenopus laevis oocytes showed that VvNPF6.5 uptake nitrate in a pH dependent way and function as a dual-affinity nitrate transporter involved in both high- and low-affinity nitrate uptake. Further ectopic expression of VvNPF6.5 in Arabidopsis resulted in more NO accumulation in shoots and roots and significantly improved nitrogen use efficiency (NUE). Moreover, VvNPF6.5 might participate in the nitrate signaling by positively regulating the expression of primary nitrate response genes.
CONCLUSION
Our results suggested that VvNPF6.5 encodes a pH-dependent, dual-affinity nitrate transporter. VvNPF6.5 regulates nitrate uptake and allocation in grapevines and is involved in primary nitrate response.
背景
硝酸盐在葡萄的营养生长和生殖生长发育中起着重要作用。然而，葡萄对硝酸盐的吸收、转运和利用方式以及其分子机制仍有待研究。
结果
本研究报告了 Vitis vinifera 硝酸盐转运蛋白 1/肽转运蛋白家族（NRT1/PTR/NPF）成员 VvNPF6.5 的功能特征。在拟南芥原生质体中的亚细胞定位表明 VvNPF6.5 定位于质膜。定量 RT-PCR 分析表明，VvNPF6.5 主要在根和茎中表达，其表达受硝酸盐的快速诱导。利用 cRNA 注射非洲爪蟾卵母细胞进行功能表征表明，VvNPF6.5 以 pH 依赖的方式摄取硝酸盐，并作为一种双亲和性硝酸盐转运体，参与高亲和性和低亲和性硝酸盐的摄取。进一步在拟南芥中异位表达 VvNPF6.5 导致地上部和根部的硝酸盐积累增加，氮素利用效率（NUE）显著提高。此外，VvNPF6.5 可能通过正向调控初级硝酸盐响应基因的表达参与硝酸盐信号转导。
结论
我们的研究结果表明，VvNPF6.5 编码一种 pH 依赖性的、双亲和性的硝酸盐转运体。VvNPF6.5 调节葡萄对硝酸盐的吸收和分配，并参与初级硝酸盐响应。
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