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反义转基因烟草植株中多酚氧化酶基因对信号分子、食草动物侵害及机械损伤响应的表达分析

Expression analysis of the polyphenol oxidase gene in response to signaling molecules, herbivory and wounding in antisense transgenic tobacco plants.

作者信息

Aziz Ejaz, Batool Riffat, Akhtar Wasim, Rehman Shazia, Gregersen Per L, Mahmood Tariq

机构信息

Department of Botany, Government Degree College Khanpur KPK, Haripur, Pakistan.

2Department of Plant Sciences, Quaid-i-Azam University, Islamabad, Pakistan.

出版信息

3 Biotech. 2019 Feb;9(2):55. doi: 10.1007/s13205-019-1587-x. Epub 2019 Jan 25.

Abstract

KEY MESSAGE

We provide evidence that the expression of the gene was significantly reduced in response to wounding, MeJ and herbivory in transgenic tobacco under wound-inducible RGLP2 promoter in an anti-sense orientation.

ABSTRACT

Polyphenol oxidase (PPO) genes play an important role in plant defense mechanisms against biotic and abiotic stresses. In the present study, a 655 bp core sequence of the potato gene was placed under the control of wound-inducible RGLP2 promoter in an anti-sense direction to evaluate its potential effects during biotic ('s infestation) and various abiotic (wounding, MeJ, ABA) stresses. Transcriptional profiling of gene by real-time PCR (qRT-PCR) in transgenic tobacco revealed a significant suppression (3.5-fold) of in response to wounding than control plants after 24 h. In response to MeJ at different concentrations (100 µM and 200 µM), the expression was greatly down-regulated by 4.7-fold after 6 h at 100 µM MeJ, and a non-significant expression was observed with ABA treatment. Moreover, significant levels of reduction (sixfolds) was found in whitefly feeding assay indicating that expression of potato in an anti-sense orientation had down-regulated the activity. This down-regulation of by wounding, MeJ and whitefly infestation clearly links the specific expression of in biotic and abiotic stresses. In the future, gene suppression in transgenic plants using anti-sense potato gene construct can be used to inhibit enzymatic browning in fruits and vegetables, e.g., potato.

摘要

关键信息

我们提供的证据表明,在伤口诱导型RGLP2启动子反义方向的转基因烟草中,该基因的表达在受到创伤、茉莉酸甲酯(MeJ)和食草动物侵害时显著降低。

摘要

多酚氧化酶(PPO)基因在植物抵御生物和非生物胁迫的防御机制中发挥重要作用。在本研究中,将马铃薯基因的一段655bp核心序列置于伤口诱导型RGLP2启动子的控制下,以反义方向评估其在生物胁迫(烟粉虱侵染)和各种非生物胁迫(创伤、茉莉酸甲酯、脱落酸(ABA))期间的潜在影响。通过实时定量聚合酶链反应(qRT-PCR)对转基因烟草中的该基因进行转录谱分析,结果显示,在24小时后,与对照植株相比,转基因烟草在受到创伤时该基因的表达显著受到抑制(3.5倍)。在不同浓度(100µM和200µM)的茉莉酸甲酯处理下,在100µM茉莉酸甲酯处理6小时后,该基因的表达大幅下调了4.7倍,而在脱落酸处理下未观察到显著的表达变化。此外,在烟粉虱取食试验中发现该基因的表达显著降低(6倍),这表明反义方向的马铃薯基因表达下调了该基因的活性。创伤、茉莉酸甲酯和烟粉虱侵染导致的该基因下调清楚地表明了该基因在生物和非生物胁迫中的特异性表达。未来,利用反义马铃薯基因构建体抑制转基因植物中的该基因,可用于抑制水果和蔬菜(如马铃薯)中的酶促褐变。

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