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拓扑异构酶II的过表达增强烟草对盐胁迫的耐受性。

Over-expression of Topoisomerase II Enhances Salt Stress Tolerance in Tobacco.

作者信息

John Riffat, Ganeshan Uma, Singh Badri N, Kaul Tanushri, Reddy Malireddy K, Sopory Sudhir K, Rajam Manchikatla V

机构信息

Plant Molecular Biology Laboratory, Department of Botany, University of Kashmir Srinagar, India.

Plant Polyamine, Transgenic and RNAi Laboratory, Department of Genetics, University of Delhi South Campus New Delhi, India.

出版信息

Front Plant Sci. 2016 Aug 31;7:1280. doi: 10.3389/fpls.2016.01280. eCollection 2016.

Abstract

Topoisomerases are unique enzymes having an ability to remove or add DNA supercoils and untangle the snarled DNA. They can cut, shuffle, and religate DNA strands and remove the torsional stress during DNA replication, transcription or recombination events. In the present study, we over-expressed topoisomerase II (TopoII) in tobacco (Nicotiana tabaccum) and examined its role in growth and development as well as salt (NaCl) stress tolerance. Several putative transgenic plants were generated and the transgene integration and expression was confirmed by PCR and Southern blot analyses, and RT-PCR analysis respectively. Percent seed germination, shoot growth, and chlorophyll content revealed that transgenic lines over-expressing the NtTopoIIα-1 gene exhibited enhanced tolerance to salt (150 and 200 mM NaCl) stress. Moreover, over-expression of TopoII lead to the elevation in proline and glycine betaine levels in response to both concentrations of NaCl as compared to wild-type. In response to NaCl stress, TopoII over-expressing lines showed reduced lipid peroxidation derived malondialdehyde (MDA) generation. These results suggest that TopoII plays a pivotal role in salt stress tolerance in plants.

摘要

拓扑异构酶是一类独特的酶,能够去除或添加DNA超螺旋,并解开缠结的DNA。它们可以切割、重排和重新连接DNA链,并在DNA复制、转录或重组过程中消除扭转应力。在本研究中,我们在烟草(Nicotiana tabaccum)中过表达拓扑异构酶II(TopoII),并研究其在生长发育以及盐(NaCl)胁迫耐受性中的作用。通过PCR和Southern杂交分析以及RT-PCR分析分别鉴定了几个推定的转基因植株,并确认了转基因的整合和表达。种子发芽率、芽生长和叶绿素含量表明,过表达NtTopoIIα-1基因的转基因株系对盐(150和200 mM NaCl)胁迫表现出增强的耐受性。此外,与野生型相比,TopoII的过表达导致在两种NaCl浓度下脯氨酸和甘氨酸甜菜碱水平升高。响应NaCl胁迫,过表达TopoII的株系显示脂质过氧化衍生的丙二醛(MDA)生成减少。这些结果表明TopoII在植物耐盐胁迫中起关键作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00e6/5006100/5cf92e56920d/fpls-07-01280-g0001.jpg

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