Laboratory of Modern Biotechnology, School of Forestry and Landscape Architecture, Anhui Agricultural University, Hefei, 230036, China.
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China.
Plant Physiol Biochem. 2021 Jul;164:205-221. doi: 10.1016/j.plaphy.2021.05.002. Epub 2021 May 12.
Plant monovalent cation/proton antiporters (CPAs), types of transmembrane transporters, play important roles in resistance to salt stress. In this study, 37 CPA genes from moso bamboo (Phyllostachys edulis) were identified and characterised. The expression profiles of 10 CPA1 genes (PheNHXs) of moso bamboo were detected by qRT-PCR, which showed that they were specifically expressed in six tissues. In addition, the expression of 10 PheNHXs in leaves and roots changed significantly under 150/200 mM NaCl and 100 μM ABA treatments. In particular, the expression of PheNHX2 in leaves and roots was significantly upregulated under NaCl treatment, thus, we cloned PheNHX2 and analysed its function. Subcellular localisation analysis showed that PheNHX2 was located on the vacuolar membrane. Overexpression of PheNHX2 reduced seed germination and root growth of Arabidopsis thaliana under salt stress, as well as severely affecting cellular Na and K content, which in turn reduced the salt tolerance of transgenic Arabidopsis. Measurements of physiological indicators, including chlorophyll content, malondialdehyde content, peroxidase and catalase enzyme activities and relative electrical conductivity, all supported this conclusion. Under salt stress, PheNHX2 also inhibited the expression of some stress-related and ion transport-related genes in transgenic Arabidopsis. Overall, these results indicate that overexpression of PheNHX2 reduces the salt tolerance of transgenic Arabidopsis. This investigation establishes a foundation for subsequent functional studies of moso bamboo CPA genes, and it provides a deeper understanding of PheNHX2 regulation in relation to the salt tolerance of moso bamboo.
植物单价阳离子/质子反向转运蛋白(CPAs)是一类跨膜转运蛋白,在耐盐胁迫中发挥重要作用。本研究从毛竹(Phyllostachys edulis)中鉴定并分析了 37 个 CPA 基因。通过 qRT-PCR 检测了毛竹 10 个 CPA1 基因(PheNHXs)的表达谱,结果表明它们在 6 种组织中特异性表达。此外,10 个 PheNHXs 在叶片和根中对 150/200 mM NaCl 和 100 μM ABA 处理的表达变化显著。特别是,PheNHX2 在叶片和根中对 NaCl 处理的表达显著上调,因此我们克隆了 PheNHX2 并分析了其功能。亚细胞定位分析表明 PheNHX2 定位于液泡膜上。过表达 PheNHX2 会降低拟南芥种子萌发和根生长对盐胁迫的耐性,严重影响细胞内 Na 和 K 含量,从而降低转基因拟南芥的耐盐性。对生理指标的测量,包括叶绿素含量、丙二醛含量、过氧化物酶和过氧化氢酶活性以及相对电导率,都支持这一结论。在盐胁迫下,PheNHX2 还抑制了转基因拟南芥中一些与应激和离子转运相关基因的表达。总之,这些结果表明过表达 PheNHX2 降低了转基因拟南芥的耐盐性。本研究为毛竹 CPA 基因的后续功能研究奠定了基础,同时也深入了解了 PheNHX2 对毛竹耐盐性的调控作用。