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Overexpression of the formaldehyde dehydrogenase gene from brevibacillus brevis to enhance formaldehyde tolerance and detoxification of tobacco

机译:短芽孢杆菌中甲醛脱氢酶基因的过表达增强烟草的甲醛耐受性和排毒

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摘要

The faldh gene coding for a putative Brevibacillus brevis formaldehyde dehydrogenase (FALDH) was isolated and then transformed into tobacco. A total of three lines of transgenic plants were generated, with each showing 2- to 3-fold higher specific formaldehyde dehydrogenase activities than wild-type tobacco, a result that demonstrates the functional activity of the enzyme in formaldehyde (HCHO) oxidation. Overexpression of faldh in tobacco confers a high tolerance to exogenous HCHO and an increased ability to take up HCHO. A 13C- nuclear magnetic resonance technique revealed that the transgenic plants were able to oxidize more aqueous HCHO to formate than the wild-type (WT) plants. When treated with gaseous HCHO, the transgenic tobacco exhibited an enhanced ability to transform more HCHO into formate, citrate acid, and malate but less glycine than the WT plants. These results indicate that the increased capacity of the transgenic tobacco to take up, tolerate, and metabolize higher concentrations of HCHO was due to the overexpression of B. brevis FALDH, revealing the essential function of this enzyme in HCHO detoxification. Our results provide a potential genetic engineering strategy for improving the phytoremediation of HCHO pollution.
机译:分离出一种编码faldh的基因,该faldh基因编码了短的短杆菌(Brevibacillus brevis)甲醛脱氢酶(FALDH),然后转化为烟草。共产生了三株转基因植物,每株都显示出比野生型烟草高2至3倍的比甲醛脱氢酶活性,这一结果证明了该酶在甲醛(HCHO)氧化中的功能活性。烟草中faldh的过度表达赋予了对外源HCHO的高度耐受性,并增加了吸收HCHO的能力。 13 C核磁共振技术表明,与野生型(WT)植物相比,转基因植物能够将更多的HCHO水溶液氧化成甲酸盐。当用气态HCHO处理时,与野生型植物相比,转基因烟草显示出将更多的HCHO转化为甲酸盐,柠檬酸和苹果酸的能力增强,但甘氨酸较少。这些结果表明,转基因烟草吸收,耐受和代谢更高浓度的HCHO的能力增强是由于短双歧杆菌FALDH的过表达,揭示了该酶在HCHO排毒中的基本功能。我们的结果为改善HCHO污染的植物修复提供了潜在的基因工程策略。

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