Division of Plant Biology, Centenary Campus, Bose Institute, Kolkata, 700054, India.
Department of Molecular Medicine, Centenary Campus, Bose Institute, Kolkata, 700054, India.
Planta. 2017 Nov;246(5):1029-1044. doi: 10.1007/s00425-017-2750-4. Epub 2017 Aug 2.
Transgenic mustard plants ( Brassica juncea ) expressing non-allergenic and biologically safe RiD peptide show higher tolerance against Lipaphis erysimi. Rorippa indica defensin (RiD) has previously been reported as a novel insecticidal protein derived from a wild crucifer Rorippa indica. RiD was found to have an effective insecticidal property against mustard aphid, Lipaphis erysimi. In the present study, RiD was highly upregulated in R. indica during aphid infestation initiating a defense system mediated by jasmonic acid (JA), but not by salicylic acid (SA)/abscisic acid (ABA). RiD has also been assessed for biosafety according to the FAO/WHO guideline (allergenicity of genetically modified foods; Food And Agriculture Organisation of the United Nations, Rome, Italy, 2001) and Codex Alimentarius Guideline (Guidelines for the design and implementation of national regulatory food safety assurance programme associated with the use of veterinary drugs in food producing animals. Codex Alimentarius Commission. GL, pp 71-2009, 2009). The purified protein was used to sensitize BALB/c mice and they showed normal histopathology of lung and no elevated IgE level in their sera. As the protein was found to be biologically safe and non-allergenic, it was used to develop transgenic Brassica juncea plants with enhanced aphid tolerance, which is one of the most important oilseed crops and is mostly affected by the devastating pest-L. erysimi. The transgene integration was monitored by Southern hybridization, and the positive B. juncea lines were further analyzed by Western blot, ELISA, immunohistolocalization assays and in planta insect bioassay. Transgenic plants expressing RiD conferred a higher level of tolerance against L. erysimi. All these results demonstrated that RiD is a novel, biologically safe, effective insecticidal agent and B. juncea plants expressing RiD are important components of integrated pest management.
表达无过敏原和生物安全 RiD 肽的转基因芥菜（Brassica juncea）对李蚜（Lipaphis erysimi）表现出更高的耐受性。先前已经报道过 RiD 是一种来自野生十字花科植物 Rorippa indica 的新型杀虫蛋白。RiD 被发现对芥菜蚜虫具有有效的杀虫特性。在本研究中，RiD 在蚜虫侵袭期间在 R. indica 中高度上调，启动了由茉莉酸（JA）介导的防御系统，但不由水杨酸（SA）/脱落酸（ABA）介导。根据粮农组织/世卫组织准则（转基因食品的变应原性；联合国粮食及农业组织，意大利罗马，2001 年）和食品法典委员会准则（与在食用动物中使用兽药相关的国家监管食品安全保证计划的设计和实施指南。食品法典委员会。GL，第 71-2009 页，2009 年），RiD 还进行了生物安全性评估。纯化的蛋白用于敏化 BALB/c 小鼠，它们的肺部组织病理学正常，血清中的 IgE 水平没有升高。由于该蛋白被发现具有生物安全性和非变应原性，因此被用于开发具有增强的蚜虫耐受性的转基因芥菜植物，芥菜是最重要的油料作物之一，主要受到毁灭性害虫-李蚜的影响。通过 Southern 杂交监测转基因的整合，并用 Western blot、ELISA、免疫组化定位分析和植物体内昆虫生物测定进一步分析阳性 B. juncea 系。表达 RiD 的转基因植物对 L. erysimi 表现出更高水平的耐受性。所有这些结果表明，RiD 是一种新型的、生物安全的、有效的杀虫剂，表达 RiD 的 B. juncea 植物是综合虫害管理的重要组成部分。
