Guan Qingjie, Wang Zhenjuan, Wang Xuhui, Takano Tetsuo, Liu Shenkui
Key Laboratory of Saline-Alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, No. 26 Hexing Road, Nangang District, Harbin 150040, China; Laboratory of Soybean Molecular Biology and Molecular Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, No. 138 Haping Road, Nangang District, Harbin 150081, China.
Key Laboratory of Saline-Alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, No. 26 Hexing Road, Nangang District, Harbin 150040, China.
J Plant Physiol. 2015 Mar 1;175:183-91. doi: 10.1016/j.jplph.2014.10.020. Epub 2014 Nov 29.
Ascorbate peroxidase (APX, EC 1.11.1.11) is one of the major members of the ROS scavenging system that plays an important role in improving saline-alkali tolerance. Puccinellia tenuiflora, as a perennial wild grass, is able to grow in extreme saline-alkali soil environments. In this study, we investigated the relationship between the P. tenuiflora ascorbate peroxidase (PutAPX) gene and saline-alkali tolerance. A phylogenetic analysis indicated that PutAPX is closely related to AtAPX3 and OsAPX4 and that these genes are on the same branch. The PutAPX-GFP fusion protein is located in the peroxisome in onion epidermal cells. The transcriptional expression of PutAPX increased with prolonged exposure to NaCl, NaHCO3, PEG6000 and H2O2 stresses in P. tenuiflora. The overexpression of PutAPX in Arabidopsis thaliana significantly increased the tolerance of plants treated with 150 and 175mM NaCl and decreased the extent of lipid peroxidation. The transgenic seedlings presented higher chlorophyll content than wild type (WT) seedlings treated with 1, 3, and 5mM NaHCO3 and 3mM H2O2. The DAB staining results revealed that the H2O2 content in transgenic seedlings was significantly lower than that in WT plants under both normal conditions and 200mM NaCl stress. Moreover, the expression of APX proteins and enzyme activity in the transgenic seedlings increased to level that were greater than twofold higher than those found in WT plants exposed to 200mM NaCl. The saline-alkali tolerance conferred by the PutAPX gene may provide a reliable basis for the use of molecular breeding techniques to improve plant tolerance and obtain a better understanding of the physiological mechanism of anti-oxidative and ROS stresses.
抗坏血酸过氧化物酶(APX,EC 1.11.1.11)是活性氧清除系统的主要成员之一,在提高耐盐碱能力方面发挥着重要作用。碱茅作为一种多年生野生草本植物,能够在极端盐碱土壤环境中生长。在本研究中,我们调查了碱茅抗坏血酸过氧化物酶(PutAPX)基因与耐盐碱能力之间的关系。系统发育分析表明,PutAPX与AtAPX3和OsAPX4密切相关,且这些基因位于同一分支上。PutAPX-GFP融合蛋白位于洋葱表皮细胞的过氧化物酶体中。在碱茅中,PutAPX的转录表达随着NaCl、NaHCO3、PEG6000和H2O2胁迫处理时间的延长而增加。在拟南芥中过表达PutAPX显著提高了经150和175mM NaCl处理的植株的耐受性,并降低了脂质过氧化程度。转基因幼苗在用1、3和5mM NaHCO3以及3mM H2O2处理时,其叶绿素含量高于野生型(WT)幼苗。DAB染色结果显示,在正常条件和200mM NaCl胁迫下,转基因幼苗中的H2O2含量均显著低于WT植株。此外,转基因幼苗中APX蛋白的表达和酶活性增加到高于暴露于200mM NaCl的WT植株两倍以上的水平。PutAPX基因赋予的耐盐碱能力可能为利用分子育种技术提高植物耐受性以及更好地理解抗氧化和活性氧胁迫的生理机制提供可靠依据。