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甘蓝血红素加氧酶基因BoHO1的异位过表达提高了拟南芥的耐盐性:蛋白质组学分析的案例研究

Ectopic over-expression of BoHO1, a cabbage heme oxygenase gene, improved salt tolerance in Arabidopsis: A case study on proteomic analysis.

作者信息

Duan Xingliang, Dai Chen, Li Zhiwei, Zhou Heng, Xiao Tianyu, Xie Yanjie, Shen Wenbiao

机构信息

College of Life Sciences, Laboratory Center of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.

College of Life Sciences, Laboratory Center of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

J Plant Physiol. 2016 Jun 1;196-197:1-13. doi: 10.1016/j.jplph.2016.02.019. Epub 2016 Mar 19.

Abstract

Plant heme oxygenase (HO) catalyzes the oxygenation of heme to biliverdin, carbon monoxide, and free iron, and is regarded as a stress-responsive protein. Here, a cabbage HO1 gene (named as BoHO1) was isolated and characterized. BoHO1 shares a high degree homology with Arabidopsis AtHO1, and could locate in Arabidopsis chloroplast. BoHO1 mRNA was ubiquitously expressed in cabbage tissues, and was responsive to several stimuli and chemicals. Genetic evidence illustrated that over-expression of BoHO1 in transgenic Arabidopsis plants (35S:BoHO1-1 and 35S:BoHO1-2) significantly alleviated salinity stress-inhibited seedling growth, which were accompanied with the re-establishment of reactive oxygen species and ion homeostasis. Comparative proteomic analysis was subsequently performed. Results revealed that protein abundance related to light reactions was greatly suppressed by NaCl stress in wild-type, whereas was partially recovered in 35S:BoHO1-1. Salinity stress also strongly activated stress-related metabolic processes in wild-type, i.e. carbon and energy metabolism, ammonium detoxification, and protein turnover, and these induced tendencies were more intensive in 35S:BoHO1-1. Particularly, proteins related to glutathione metabolism and ion homeostasis were specifically enriched in NaCl-stressed 35S:BoHO1-1. On the basis of above results, we propose that BoHO1 could activate multiple stress-responsive pathways to help Arabidopsis regain cellular homeostasis, thus presenting enhanced adaptation to salinity stress.

摘要

植物血红素加氧酶(HO)催化血红素氧化生成胆绿素、一氧化碳和游离铁,被认为是一种应激反应蛋白。在此,分离并鉴定了一个甘蓝HO1基因(命名为BoHO1)。BoHO1与拟南芥AtHO1具有高度同源性,且可定位于拟南芥叶绿体中。BoHO1 mRNA在甘蓝组织中普遍表达,并对多种刺激和化学物质有反应。遗传学证据表明,在转基因拟南芥植株(35S:BoHO1-1和35S:BoHO1-2)中过表达BoHO1可显著缓解盐胁迫对幼苗生长的抑制,这伴随着活性氧和离子稳态的重新建立。随后进行了比较蛋白质组学分析。结果显示,野生型中与光反应相关的蛋白质丰度在NaCl胁迫下受到极大抑制,而在35S:BoHO1-1中部分恢复。盐胁迫还强烈激活了野生型中与胁迫相关的代谢过程,即碳和能量代谢、铵解毒和蛋白质周转,且这些诱导趋势在35S:BoHO1-1中更为强烈。特别地,与谷胱甘肽代谢和离子稳态相关的蛋白质在NaCl胁迫下的35S:BoHO1-1中特异性富集。基于上述结果,我们提出BoHO1可激活多种应激反应途径,帮助拟南芥恢复细胞稳态,从而增强对盐胁迫的适应性。

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