Red fluorescent protein (DsRed2), an ideal reporter for cotton genetic transformation and molecular breeding
Red fluorescent protein (DsRed2), an ideal reporter for cotton genetic transformation and molecular breeding
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DOI：
10.1016/j.cj.2018.05.002
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发表时间：
2018-08-01
期刊：
影响因子：
6.6
通讯作者：
Jin, Shuangxia
中科院分区：
文献类型：
--
作者：
Sun, Lin;Alariqi, Muna;Jin, Shuangxia
编码报告蛋白的基因在遗传转化和植物育种中被用作视觉标记辅助工具。本研究成功地将珊瑚Discosoma sp. coral中鉴定的红色荧光蛋白（DsRed2）作为棉花基因工程的视觉标记。在CaMV-35S启动子的驱动下，通过农杆菌介导的转化，DsRed2在棉花品种JIN668和YZ1中成功表达。我们的结果表明，DsRed2的表达通过视觉观察为转基因愈伤组织的早期选择提供了一个工具。红色荧光不仅可以在愈伤组织和体细胞胚中检测到，而且可以在成熟植物的大多数组织和器官中检测到。将转基因品系Yz-2-DsRed2与4个不同棉花品种杂交，在不同遗传背景下评价转基因的遗传力和稳定性。以Yz-2-DsRed2为父本时，红色遗传力高度稳定。DsRed2基因在F1杂交种中100%表达。为了研究DsRed2转录与DNA甲基化之间的关系，我们对CaMV-35S启动子区域采用了甲基化特异性PCR方法。结果表明，启动子区DNA甲基化水平与转基因转录呈负相关。综上所述，这些发现表明DsRed2是棉花遗传转化和分子育种计划的视觉报告基因。(C) 2018年中国作物科学学会，中国农业科学院作物科学研究所。由Elsevier B.V.代表KeAi Communications Co. Ltd制作和主持。
Genes encoding reporter proteins are used as visual marker-assisted tools in genetic transformation as well as plant breeding. In this study, the red fluorescent protein identified in Discosoma sp. coral (DsRed2) was successfully used as a visual marker for cotton genetic engineering. DsRed2 was successfully expressed in two cotton cultivars, JIN668 and YZ1, driven by the CaMV-35S promoter via the Agrobacterium-mediated transformation. Our results suggest that DsRed2 expression provides an early-stage selection tool for the transgenic calli via visual observation. Red fluorescence can be detected not only in callus and somatic embryos but also in most tissues and organs of mature plants. The transgenic line Yz-2-DsRed2 was crossed with four different cotton cultivars to assess the transgene heritability and stability in different genetic backgrounds. The heritability of the red color was highly stable when Yz-2-DsRed2 was used as a male parent. The DsRed2 gene expressed 100% in the F1 hybrids. To investigate the relationship between DsRed2 transcription and DNA methylation, a methylation-specific PCR approach was applied to the CaMV-35S promoter region. The results showed a negative association between DNA methylation level in the promoter region and the transgene transcription. Taken together, these findings suggest DsRed2 a visual reporter gene for cotton genetic transformation and molecular breeding programs. (C) 2018 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.