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快速且高效的形态发生基因介导的六倍体小麦转化

Rapid and highly efficient morphogenic gene-mediated hexaploid wheat transformation.

作者信息

Johnson Kari, Cao Chu Uyen, Anthony Geny, Wu Emily, Che Ping, Jones Todd J

机构信息

Corteva Agriscience, Johnston, IA, United States.

出版信息

Front Plant Sci. 2023 Mar 29;14:1151762. doi: 10.3389/fpls.2023.1151762. eCollection 2023.

Abstract

The successful employment of morphogenic regulator genes, () and (), for -mediated transformation of maize ( L.) and sorghum ( L.) has been reported to improve transformation by inducing rapid somatic embryo formation. Here, we report two morphogenic gene-mediated wheat transformation methods, either with or without morphogenic and marker gene excision. These methods yield independent-transformation efficiency up to 58% and 75%, respectively. In both cases, the tissue culture duration for generating transgenic plants was significantly reduced from 80 to nearly 50 days. In addition, the transformation process was significantly simplified to make the procedure less labor-intensive, higher-throughput, and more cost-effective by eliminating the requirement for embryonic axis excision, bypassing the necessity for prolonged dual-selection steps for callus formation, and obviating the prerequisite of cytokinin for shoot regeneration. Furthermore, we have demonstrated the flexibility of the methods and generated high-quality transgenic events across multiple genotypes using herbicide (phosphinothricin, ethametsulfuron)- and antibiotic (G418)-based selections.

摘要

据报道,形态发生调节基因()和()成功用于介导玉米(L.)和高粱(L.)的转化,通过诱导快速体细胞胚胎形成来提高转化效率。在此,我们报告了两种形态发生基因介导的小麦转化方法,一种有形态发生和标记基因切除,另一种没有。这些方法的独立转化效率分别高达58%和75%。在这两种情况下,产生转基因植物的组织培养时间从80天显著缩短至近50天。此外,通过消除胚胎轴切除的要求、绕过愈伤组织形成所需的长时间双重选择步骤以及避免芽再生所需的细胞分裂素,转化过程得到了显著简化,从而使该程序劳动强度更低、通量更高且更具成本效益。此外,我们已经证明了这些方法的灵活性,并使用基于除草剂(草铵膦、乙磺隆)和抗生素(G418)的选择,在多个基因型中产生了高质量的转基因事件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1660/10090459/f20d94b4cc75/fpls-14-1151762-g001.jpg

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