Khan Muhammad Sarwar, Kanwal Benish, Nazir Shahid
Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, Faisalabad Pakistan.
National Institute for Biotechnology and Genetic Eningeering, Faisalabad Pakistan.
Front Plant Sci. 2015 Sep 9;6:725. doi: 10.3389/fpls.2015.00725. eCollection 2015.
Osmoprotectants stabilize proteins and membranes against the denaturing effect of high concentrations of salts and other harmful solutes. In yeast, arabitol dehydrogenase (ArDH) reduces D-ribulose to D-arabitol where D-ribulose is derived by dephosphorylating D-ribulose-5-PO4 in the oxidized pentose pathway. Osmotolerance in plants could be developed through metabolic engineering of chloroplast genome by introducing genes encoding polyols since chloroplasts offer high level transgene expression and containment. Here, we report that ArDH expression in tobacco chloroplasts confers tolerance to NaCl (up to 400 mM). Transgenic plants compared to wild type (WT) survived for only 4-5 weeks on 400 mM NaCl whereas plants remained green and grew normal on concentrations up to 350 mM NaCl. Further, a-week-old seedlings were also challenged with poly ethylene glycol (PEG, up to 6%) in the liquid medium, considering that membranes and proteins are protected under stress conditions due to accumulation of arabitol in chloroplasts. Seedlings were tolerant to 6% PEG, suggesting that ARDH enzyme maintains integrity of membranes in chloroplasts under drought conditions via metabolic engineering. Hence, the gene could be expressed in agronomic plants to withstand abiotic stresses.
渗透保护剂可稳定蛋白质和膜,使其免受高浓度盐及其他有害溶质的变性作用。在酵母中,阿拉伯糖醇脱氢酶(ArDH)将D-核糖还原为D-阿拉伯糖醇,其中D-核糖是通过氧化戊糖途径中D-核糖-5-磷酸的去磷酸化作用产生的。由于叶绿体具有高水平的转基因表达和隔离作用,通过导入编码多元醇的基因对叶绿体基因组进行代谢工程改造,可以提高植物的耐渗性。在此,我们报道烟草叶绿体中ArDH的表达赋予了对NaCl(高达400 mM)的耐受性。与野生型(WT)相比,转基因植物在400 mM NaCl上仅存活4 - 5周,而在高达350 mM NaCl的浓度下,植物仍保持绿色并正常生长。此外,考虑到由于叶绿体中阿拉伯糖醇的积累,膜和蛋白质在胁迫条件下受到保护,一周龄的幼苗也在液体培养基中用聚乙二醇(PEG,高达6%)进行处理。幼苗对6% PEG具有耐受性,这表明通过代谢工程,ARDH酶在干旱条件下维持叶绿体中膜的完整性。因此,该基因可以在农艺植物中表达以抵御非生物胁迫。