用于大豆基因组编辑视觉选择的 RUBY 报告器

IF 4.6 4区 农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Li Chen, Yupeng Cai, Xiaoqian Liu, Weiwei Yao, Shuiqing Wu, Wensheng Hou
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引用次数: 0

摘要

目前筛选转基因大豆(Glycine max)的系统涉及费力的分子检测或荧光标记的表达,而荧光标记在大豆植株中很难看到。因此,用于早期筛选转基因植物的可视系统将提高通过基因组编辑改良作物的效率。RUBY 报告系统由三个编码甜菜红素生物合成酶的基因组成,可导致转基因组织中紫色色素的积累,已在一些植物和二核真菌中使用。在此,我们对 RUBY 报告物进行了评估,以便在大豆转化过程中进行视觉验证。我们的研究表明,RUBY 可以在大豆中表达,从而无需专门设备即可对转基因事件进行视觉确认。在任何组织中都能看到紫色色素积累的植株被成功转化,这证实了 RUBY 系统作为视觉指标的准确性。我们还利用后代的子叶评估了转基因在不同世代间的遗传稳定性。不含转基因的幼苗具有明显的绿色,便于选择经过基因组编辑但不含转基因的大豆幼苗进行收获。利用 RUBY 系统,我们很快就在 T1 代中鉴定出了无转基因的 Gmwaxy 突变体。因此,该系统为大豆基因组编辑提供了一种高效便捷的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The RUBY reporter for visual selection in soybean genome editing

Current systems to screen for transgenic soybeans (Glycine max) involve laborious molecular assays or the expression of fluorescent markers that are difficult to see in soybean plants. Therefore, a visual system for early screening of transgenic plants would increase the efficiency of crop improvement by genome editing. The RUBY reporter system, which consists of three genes encoding betalain biosynthetic enzymes, leading to the accumulation of purple pigment in transgenic tissue, has been employed in some plants and dikaryon fungi. Here, we assessed the RUBY reporter for visual verification during soybean transformation. We show that RUBY can be expressed in soybean, allowing for visual confirmation of transgenic events without the need for specialized equipment. Plants with visible accumulation of purple pigment in any tissue were successfully transformed, confirming the accuracy of the RUBY system as a visual indicator. We also assessed the genetic stability of the transgene across generations, which can be performed very early, using the cotyledons of the progeny. Transgene-free seedlings have a distinct green color, facilitating the selection of genome-edited but transgene-free soybean seedlings for harvest. Using the RUBY system, we quickly identified a transgene-free Gmwaxy mutant in the T1 generation. This system thus provides an efficient and convenient tool for soybean genome editing.

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CiteScore
7.70
自引率
2.80%
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