Overexpression of a specific soybean GmGSTU4 isoenzyme improves diphenyl ether and chloroacetanilide herbicide tolerance of transgenic tobacco plants
Overexpression of a specific soybean GmGSTU4 isoenzyme improves diphenyl ether and chloroacetanilide herbicide tolerance of transgenic tobacco plants
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DOI：
10.1016/j.jbiotec.2010.07.011
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发表时间：
2010-10-01
影响因子：
4.1
通讯作者：
Tsaftaris, Athanasios
中科院分区：
文献类型：
--
作者：
Benekos, Kostantinos;Kissoudis, Christos;Tsaftaris, Athanasios
植物谷胱甘肽转移酶（GSTs）超家族是一类多功能酶，是植物除草剂解毒酶网络的重要组成部分。来自大豆的tau类GST同工酶GmGSTU 4对二苯醚除草剂氟草醚表现出催化活性，并且作为谷胱甘肽依赖性过氧化物酶（GPOX）具有活性。通过农杆菌转化产生Basmas栽培品种的转基因烟草植物。目的是评估在植物中，GmGSTU 4的作用，在解毒二苯醚除草剂氟草醚和乙氧氟草醚和氯乙酰苯胺甲草胺和异丙甲草胺。通过PCR和Southern印迹杂交对转基因烟草植株进行了验证，并通过RT-PCR检测了GmGSTU 4的表达。转基因植物的叶提取物显示出对CDNB的GST活性适度增加，对氟草醚和甲草胺的GST活性显著增加，同时对氢过氧化枯烯的GPOX活性增加。当用200 μ M氟草醚或乙氧氟草醚处理时，与野生型植物相比，GmGSTU 4过表达植物表现出降低的相对电解质渗漏。此外，与野生型植物相比，当在7.5 mg/L甲草胺下体外生长时，所有GmGSTU 4过表达株系表现出对甲草胺的耐受性显著增加。未观察到对异丙甲草胺的耐受性显著增加。这些结果证实了这种特殊的GmGSTU 4同工酶从大豆中的氟草醚和甲草胺的解毒的贡献，并提供了基础，对转基因植物的发展与改善植物修复能力，为未来用于环境净化除草剂。(c)2010爱思唯尔有限公司版权所有。
Plant glutathione transferases (GSTs) superfamily consists of multifunctional enzymes and forms a major part of the plants herbicide detoxification enzyme network. The tau class GST isoenzyme GmGSTU4 from soybean, exhibits catalytic activity towards the diphenyl ether herbicide fluorodifen and is active as glutathione-dependent peroxidase (GPOX). Transgenic tobacco plants of Basmas cultivar were generated via Agrobacterium transformation. The aim was to evaluate in planta, GmGSTU4's role in detoxifying the diphenyl ether herbicides fluorodifen and oxyfluorfen and the chloroacetanilides alachlor and metolachlor. Transgenic tobacco plants were verified by PCR and Southern blot hybridization and expression of GmGSTU4 was determined by RT-PCR. Leaf extracts from transgenic plants showed moderate increase in GST activity towards CDNB and a significant increase towards fluorodifen and alachlor, and at the same time an increased GPOX activity towards cumene hydroperoxide. GmGSTU4 overexpressing plants when treated with 200 mu M fluorodifen or oxyfluorfen exhibited reduced relative electrolyte leakage compared to wild type plants. Moreover all GmGSTU4 overexpressing lines exhibited significantly increased tolerance towards alachlor when grown in vitro at 7.5 mg/L alachlor compared to wild type plants. No significant increased tolerance was observed to metolachlor. These results confirm the contribution of this particular GmGSTU4 isoenzyme from soybean in the detoxification of fluorodifen and alachlor, and provide the basis towards the development of transgenic plants with improved phytoremediation capabilities for future use in environmental cleanup of herbicides. (c) 2010 Elsevier B.V. All rights reserved.