Oilseeds Research Station, Tamil Nadu Agricultural University, Tindivanam 604 002, India.
Biochimie. 2010 Oct;92(10):1321-9. doi: 10.1016/j.biochi.2010.06.009. Epub 2010 Jul 1.
Plant growth and productivity are adversely affected by various abiotic stress factors. In our previous study, we used Avicennia marina, a halophytic mangrove, as a model plant system for isolating genes functioning in salt stress tolerance. A large scale random EST sequencing from a salt stressed leaf tissue cDNA library of one month old A. marina plants resulted in identification of a clone showing maximum homology to Monodehydroascorbate reductase (Am-MDAR). MDAR plays a key role in regeneration of ascorbate from monodehydroascorbate for ROS scavenging. In this paper, we report the cellular localization and the ability to confer salt stress tolerance in transgenic tobacco of this salt inducible Am-MDAR. A transit peptide at the N-terminal region of Am-MDAR suggested that it encodes a chloroplastic isoform. The chloroplastic localization was confirmed by stable transformation and expression of the Am-MDAR-GFP fusion protein in tobacco. Transgenic tobacco plants overexpressing Am-MDAR survived better under conditions of salt stress compared to untransformed control plants. Assays of enzymes involved in ascorbate-glutathione cycle revealed an enhanced activity of MDAR and ascorbate peroxidase whereas the activity of dehyroascorbate reductase was reduced under salt stressed and unstressed conditions in Am-MDAR transgenic lines. The transgenic lines showed an enhanced redox state of ascorbate and reduced levels of malondialdehyde indicating its enhanced tolerance to oxidative stress. The results of our studies could be used as a starting point for genetic engineering of economically important plants tolerant to salt stress.
植物的生长和生产力会受到各种非生物胁迫因素的不利影响。在我们之前的研究中,我们使用了盐生红树林植物海桑(Avicennia marina)作为模式植物系统,以分离在盐胁迫耐受中起作用的基因。从一个月大的海桑叶片组织 cDNA 文库中对盐胁迫进行大规模随机 EST 测序,结果鉴定出一个与单脱氢抗坏血酸还原酶(Am-MDAR)最大同源的克隆。MDAR 在从单脱氢抗坏血酸再生抗坏血酸以清除 ROS 方面发挥关键作用。在本文中,我们报告了这种盐诱导的 Am-MDAR 的细胞定位和在转基因烟草中赋予耐盐性的能力。Am-MDAR N 端区域的转运肽提示它编码一种质体同工型。通过稳定转化和 Am-MDAR-GFP 融合蛋白在烟草中的表达,证实了质体的定位。与未转化的对照植物相比,过表达 Am-MDAR 的转基因烟草在盐胁迫条件下更好地存活。参与抗坏血酸-谷胱甘肽循环的酶的测定表明,在盐胁迫和非胁迫条件下,MDAR 和抗坏血酸过氧化物酶的活性增强,而脱氢抗坏血酸还原酶的活性降低。转基因系表现出增强的抗坏血酸氧化还原状态和降低的丙二醛水平,表明其对氧化应激的耐受性增强。我们研究的结果可作为遗传工程经济上重要的耐盐植物的起点。