Exploring an economic and highly efficient genetic transformation and genome-editing system for radish through developmental regulators and visible reporter.

IF 6.2 1区 生物学 Q1 PLANT SCIENCES
Xiaofang Yi, Congcong Wang, Xiaoqi Yuan, Mi Zhang, Changwei Zhang, Tiaojiao Qin, Haiyun Wang, Liang Xu, Liwang Liu, Yan Wang
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Abstract

Radish (Raphanus sativus L.) is one of the most important root vegetable crops worldwide. However, gene function exploration and germplasm innovation still face tremendous challenges due to its extremely low transformation efficiency. Here, an economic and highly efficient genetic transformation method for radish was explored by Agrobacterium rhizogenes-mediated transformation with the help of combining special developmental regulator (DR) genes and the visual identification reporter. Firstly, the RUBY gene, a betalain biosynthesis system, could result in a visual red-violet color used as a convenient and effective reporter for monitoring transgenic hairy roots screening of radish. However, the hairy roots-to-shoots conversion system of radish still stands as a barrier to the obtainment of whole transgenic plants, although different hormone combinations and various culture conditions were tried. Following, two DR genes including Wuschel2 (Wus2) and isopentenyl transferase (ipt), as well as their combination Wus2-ipt were introduced for the shoot regeneration capacity improvement. The results showed that the transgenic shoots could be directly generated without externally supplying any hormones in the presence of a Wus2-ipt combination. Then, Wus2-ipt along with the RUBY reporter was employed to establish an efficient genetic transformation system of radish. Moreover, this system was applied in generating gene-edited radish plants and the phytoene desaturase (RsPDS) gene was effectively knockout through albino phenotype observation and sequencing analysis. These findings have the potential to be widely applied in genetic transformation and genome-editing genetic improvement of other vegetable species.

通过发育调节剂和可见报告基因,探索经济高效的萝卜遗传转化和基因组编辑系统。
萝卜(Raphanus sativus L.)是世界上最重要的根茎蔬菜作物之一。然而,由于其转化效率极低,基因功能探索和种质创新仍面临巨大挑战。本文通过根瘤农杆菌介导的转化,结合特殊的发育调节因子(DR)基因和视觉识别报告基因,探索了一种经济、高效的萝卜遗传转化方法。首先,RUBY 基因是一种甜菜碱生物合成系统,可产生视觉上的红紫色,是监测萝卜转基因毛细根筛选的一种方便有效的报告基因。然而,尽管尝试了不同的激素组合和各种培养条件,萝卜毛细根到根的转化系统仍然是获得完整转基因植株的障碍。为了提高萝卜芽的再生能力,研究人员引入了两个 DR 基因,包括 Wuschel2(Wus2)和异戊烯基转移酶(ipt),以及它们的组合 Wus2-ipt。结果表明,在 Wus2-ipt组合存在的情况下,无需外部提供任何激素就能直接产生转基因芽。随后,Wus2-ipt 与 RUBY 报告基因一起被用于建立一个高效的萝卜遗传转化系统。此外,该系统还被应用于生成基因编辑萝卜植株,通过白化表型观察和测序分析,植二烯去饱和酶(RsPDS)基因被有效敲除。这些发现有望广泛应用于其他蔬菜物种的遗传转化和基因组编辑遗传改良。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
The Plant Journal
The Plant Journal 生物-植物科学
CiteScore
13.10
自引率
4.20%
发文量
415
审稿时长
2.3 months
期刊介绍: Publishing the best original research papers in all key areas of modern plant biology from the world"s leading laboratories, The Plant Journal provides a dynamic forum for this ever growing international research community. Plant science research is now at the forefront of research in the biological sciences, with breakthroughs in our understanding of fundamental processes in plants matching those in other organisms. The impact of molecular genetics and the availability of model and crop species can be seen in all aspects of plant biology. For publication in The Plant Journal the research must provide a highly significant new contribution to our understanding of plants and be of general interest to the plant science community.
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