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大豆基因组编辑中视觉选择的报告基因。

The reporter for visual selection in soybean genome editing.

作者信息

Chen Li, Cai Yupeng, Liu Xiaoqian, Yao Weiwei, Wu Shuiqing, Hou Wensheng

机构信息

State Key Laboratory of Crop Gene Resources and Breeding, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081 China.

Ministry of Agriculture Key Laboratory of Soybean Biology (Beijing), Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081 China.

出版信息

aBIOTECH. 2024 Mar 18;5(2):209-213. doi: 10.1007/s42994-024-00148-6. eCollection 2024 Jun.

Abstract

Current systems to screen for transgenic soybeans () 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 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 reporter for visual verification during soybean transformation. We show that 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 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 system, we quickly identified a transgene-free mutant in the T1 generation. This system thus provides an efficient and convenient tool for soybean genome editing.

摘要

当前用于筛选转基因大豆的系统涉及繁琐的分子检测或在大豆植株中难以观察到的荧光标记表达。因此,一种用于早期筛选转基因植物的可视化系统将通过基因组编辑提高作物改良的效率。由三个编码甜菜碱生物合成酶的基因组成的报告系统,可导致转基因组织中积累紫色色素,已在一些植物和双核真菌中得到应用。在此,我们评估了该报告系统在大豆转化过程中的可视化验证效果。我们发现该系统能够在大豆中表达,无需专门设备即可实现对转基因事件的可视化确认。在任何组织中可见紫色色素积累的植株已成功转化,证实了该报告系统作为可视化指标的准确性。我们还评估了转基因在后代中的遗传稳定性,这可以使用后代的子叶在早期进行。无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无转基因的大豆幼苗进行收获。利用该报告系统,我们在T1代中快速鉴定出了一个无转基因的突变体。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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b730/11224211/d35882b3161b/42994_2024_148_Fig1_HTML.jpg

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