Department of Cell and Molecular Biology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
J Plant Physiol. 2012 Feb 15;169(3):311-8. doi: 10.1016/j.jplph.2011.09.004. Epub 2011 Oct 22.
Monodehydroascorbate reductase (MDHAR), an important enzyme of the ascorbate-glutathione cycle, is involved in salt tolerance of plants through scavenging of reactive oxygen species (ROS). In this study, a cDNA encoding MDHAR from the mangrove plant Acanthus ebracteatus was introduced into rice to examine its role in salt tolerance. Three stable transgenic lines (MT22, MT24 and MT25) overexpressing AeMDHAR were selected in vitro using hygromycin and confirmed by PCR, quantitative reverse-transcription (qRT) PCR and enzyme assay. The transgenic line MT24 was predicted to possess a single copy of the transgene while the other two transgenic lines were predicted to have multiple transgene integrations. The AeMDHAR transcripts were detected only in transgenic rice lines but not in untransformed rice. The abundance of AeMDHAR transcripts in transgenic lines MT22 and MT25 was approximately 2.75 times the amount found in MT24. The transgenic rice lines overexpressing AeMDHAR showed a significant increase in MDHAR enzyme activity compared to untransformed plants under both NaCl and control conditions. All transgenic lines showed better yield attributes such as a higher tiller number and increased 1000-grain weight compared to non-transgenics. They also showed tolerance to salt at germination and seedling stages. The transgenic line MT24, which harbors a single copy of AeMDHAR, displayed a lower rate of sterility, a higher number of tillers and longer panicle compared to untransformed plants when subjected to salt stress.
单脱氢抗坏血酸还原酶(MDHAR)是抗坏血酸-谷胱甘肽循环中的一种重要酶,通过清除活性氧(ROS)参与植物的耐盐性。本研究从红树植物 Acanthus ebracteatus 中引入编码 MDHAR 的 cDNA 到水稻中,以研究其在耐盐性中的作用。通过潮霉素体外选择,获得了三个稳定的转 AeMDHAR 基因株系(MT22、MT24 和 MT25),并通过 PCR、定量反转录(qRT)PCR 和酶活性测定进行了验证。预测转 AeMDHAR 的 MT24 株系含有一个单拷贝的转基因,而另外两个株系预测有多个转基因整合。在转基因水稻株系中检测到 AeMDHAR 转录本,但在未转化的水稻中未检测到。MT22 和 MT25 中的 AeMDHAR 转录本的丰度约为 MT24 中的 2.75 倍。与对照条件下相比,在盐胁迫下,过表达 AeMDHAR 的转基因水稻株系的 MDHAR 酶活性显著增加。与非转基因相比,所有转基因株系的分蘖数和千粒重等产量性状均得到改善,在发芽和幼苗阶段也表现出耐盐性。与未转化的植物相比,含有 AeMDHAR 单拷贝的转基因株系 MT24 在受到盐胁迫时,不育率较低,分蘖数较多,穗长较长。