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在细胞质和叶绿体中积累苜蓿醛糖还原酶的转基因大麦的胁迫耐受性

Stress tolerance of transgenic barley accumulating the alfalfa aldose reductase in the cytoplasm and the chloroplast.

作者信息

Nagy Bettina, Majer Petra, Mihály Róbert, Pauk János, Horváth Gábor V

机构信息

Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.

Cereal Research Non-Profit Company, Alsó kikötő sor 9, 6726, Szeged, Hungary.

出版信息

Phytochemistry. 2016 Sep;129:14-23. doi: 10.1016/j.phytochem.2016.07.007. Epub 2016 Jul 25.

DOI:10.1016/j.phytochem.2016.07.007
PMID:27469099
Abstract

Barley represents one of the major crops grown worldwide; its genetic transformation provides an important tool for the improvement of crop quality and tolerance to environmental stress factors. Biotic and abiotic stresses produce reactive oxygen species in the plant cells that can directly oxidize the cellular components including lipid membranes; resulting in lipid peroxidation and subsequently the accumulation of reactive carbonyl compounds. In order to protect barley plants from the effects of stress-produced reactive carbonyls, an Agrobacterium-mediated transformation was carried out using the Medicago sativa aldose reductase (MsALR) gene. In certain transgenic lines the produced MsALR enzyme was targeted to the chloroplasts to evaluate its protective effect in these organelles. The dual fluorescent protein-based method was used for the evaluation of tolerance of young seedlings to diverse stresses; our results demonstrated that this technique could be reliably applied for the detection of cellular stress in a variety of conditions. The chlorophyll and carotenoid content measurements also supported the results of the fluorescent protein-based method and the stress-protective effect of the MsALR enzyme. Targeting of MsALR into the chloroplast has also resulted in increased stress tolerance, similarly to the observed effect of the cytosolic MsALR accumulation. The results of the DsRed/GFP fluorescent protein-based method indicated that both the cytosol and chloroplast accumulation of MsALR can increase the abiotic stress tolerance of transgenic barley lines.

摘要

大麦是全球种植的主要作物之一;其遗传转化为提高作物品质和增强对环境胁迫因子的耐受性提供了重要工具。生物和非生物胁迫会在植物细胞中产生活性氧,这些活性氧可直接氧化包括脂质膜在内的细胞成分;导致脂质过氧化,进而使活性羰基化合物积累。为了保护大麦植株免受胁迫产生的活性羰基化合物的影响,利用紫花苜蓿醛糖还原酶(MsALR)基因进行了农杆菌介导的转化。在某些转基因株系中,所产生的MsALR酶被靶向到叶绿体中,以评估其在这些细胞器中的保护作用。基于双荧光蛋白的方法用于评估幼苗对多种胁迫的耐受性;我们的结果表明,该技术可可靠地应用于检测各种条件下的细胞胁迫。叶绿素和类胡萝卜素含量的测量结果也支持了基于荧光蛋白的方法的结果以及MsALR酶的胁迫保护作用。将MsALR靶向到叶绿体中也导致胁迫耐受性增加,这与胞质中MsALR积累所观察到的效果类似。基于DsRed/GFP荧光蛋白的方法的结果表明,MsALR在胞质和叶绿体中的积累均可提高转基因大麦株系的非生物胁迫耐受性。

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