Liu D, Liu Y, Rao J, Wang G, Li H, Ge F, Chen C
Mol Biol (Mosk). 2013 Jul-Aug;47(4):591-601. doi: 10.7868/s0026898413040101.
Glutathione S-transferases (GSTs) are ubiquitous enzymes in animals and plants, and they are multifunctional proteins encoded by a large gene family. GSTs are involved in response to the oxidative stress including drought, salt, heavy metals, and so on. Under oxidative stress, the excessive reactive oxygen species (ROS) induce an increase in GST levels, and then the GSTs metabolize the toxic products of lipid peroxidation, damaged DNA and other molecules. Previously, a full-length cDNA of a novel zeta GST gene, PpGST, was characterized from fruit of Pyrus pyrifolia Nakai cv Huobali. In the present study, a constitutive plant expression vector of PpGSTwas constructed and transferred into tobacco (Nicotiana tabacum L. cv Xanthi) to verify the function of PpGST. As a result, the PpGSTgene was successfully integrated into the genome of the transgenic tobacco lines and expressed as expected in the transformants through Southern blotting and quantitative reverse transcription-polymerase chain reaction analysis. Growth of T1 generation plants of PpGST transgenic lines and WT under non-stressful conditions was similar, however, the transgenic tobacco lines showed relatively normal growth under drought, NaCl, and cadmium (Cd) stresses. Furthermore, the T1 transgenic tobacco lines showed significantly slower superoxide anion production rate than the WT under abiotic stress. Simultaneously, the MDA content of each T1 transgenic tobacco plant was only slightly increased and significantly lower than that of the WT under drought, salt and Cd stress. Together with the GST activity of the transgenic tobacco lines, which was significantly increased under stressful conditions, as compared with that in WT, overexpression of PpGSTin tobacco enhanced the tolerance of transgenic tobacco lines to oxidative damage caused by drought, NaCl, and Cd stresses.
谷胱甘肽S-转移酶(GSTs)是动植物中普遍存在的酶,它们是由一个大基因家族编码的多功能蛋白质。GSTs参与对包括干旱、盐、重金属等在内的氧化应激反应。在氧化应激下,过量的活性氧(ROS)会导致GST水平升高,然后GSTs代谢脂质过氧化的有毒产物、受损的DNA和其他分子。此前,从火巴梨(Pyrus pyrifolia Nakai cv Huobali)果实中鉴定出一个新型ζ类GST基因PpGST的全长cDNA。在本研究中,构建了PpGST的组成型植物表达载体并将其转入烟草(Nicotiana tabacum L. cv Xanthi)以验证PpGST的功能。结果,通过Southern杂交和定量逆转录-聚合酶链反应分析,PpGST基因成功整合到转基因烟草株系的基因组中,并在转化体中如预期表达。PpGST转基因株系的T1代植株和野生型在非胁迫条件下的生长相似,然而,转基因烟草株系在干旱、NaCl和镉(Cd)胁迫下表现出相对正常的生长。此外,在非生物胁迫下,T1转基因烟草株系的超氧阴离子产生速率明显比野生型慢。同时,在干旱、盐和Cd胁迫下,每个T1转基因烟草植株的丙二醛含量仅略有增加,且显著低于野生型。与野生型相比,转基因烟草株系在胁迫条件下的GST活性显著增加,PpGST在烟草中的过表达增强了转基因烟草株系对干旱、NaCl和Cd胁迫引起的氧化损伤的耐受性。