DipTRANS: an improved method for in planta transformation and genome engineering in Nicotiana benthamiana.

IF 16.6 1区工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Trends in biotechnology Pub Date : 2026-09-12 DOI:10.1016/j.tibtech.2026.08.013

Fazhan Qi, Weiwei Chen, Jon P Cody, Hao Cheng, Jiayi Gao, James C Chamness, Yumeng Jiang, Fang Wei, Daniel F Voytas, Gongyao Shi

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引用次数: 0

Abstract

Plant transformation remains constrained by labor-intensive tissue culture. Our previous work showed that direct delivery of developmental regulators (DRs) can induce de novo meristems on plants, offering a promising transformation approach. In this resource article, we introduced DipTRANS (Direct in planta Transformation), an optimized, soil-based heritable transformation platform for Nicotiana benthamiana that bypasses sterile culture entirely. DipTRANS is built on DR-induced de novo meristem formation. After optimizing parameters, including regulator combinations, Agrobacterium strain, and infiltration density, DipTRANS yielded transformation efficiencies to 46.7%. Developmental abnormalities associated with regulator expression are resolved through cutting-based propagation and virus-induced transgene excision, enabling recovery of fertile, transgenic progeny. Furthermore, DipTRANS supports tissue culture-free, transgene-free iterative genome modification via virus-induced genome editing. Overall, DipTRANS enables the generation of transgenic plants within 30 days and engineered progeny within 90 days. This methodology provides a rapid, versatile platform and a blueprint for extending direct in planta transformation to other plant species.

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DipTRANS：一种改良的烟草植物转化和基因组工程方法。

植物转化仍然受到劳动密集型组织培养的限制。我们之前的研究表明，直接递送发育调节因子（DRs）可以诱导植物新生分生组织，这是一种很有前途的转化方法。在这篇资源文章中，我们介绍了DipTRANS (Direct In planta Transformation)，这是一个优化的、基于土壤的可遗传转化平台，完全绕过了无菌培养。DipTRANS是建立在dr诱导的新生分生组织形成上的。优化调控剂组合、农杆菌菌株和浸润密度等参数后，DipTRANS转化效率达到46.7%。通过基于切割的繁殖和病毒诱导的转基因切除，解决了与调节因子表达相关的发育异常，使可育的转基因后代得以恢复。此外，DipTRANS支持通过病毒诱导的基因组编辑进行无组织培养、无转基因的迭代基因组修饰。总的来说，DipTRANS可以在30天内产生转基因植物，并在90天内产生工程后代。该方法提供了一个快速、通用的平台，并为直接将植物转化扩展到其他植物物种提供了蓝图。

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

Trends in biotechnology 工程技术-生物工程与应用微生物

CiteScore

28.60

自引率

1.20%

发文量

198

审稿时长

1 months

期刊介绍： Trends in Biotechnology publishes reviews and perspectives on the applied biological sciences, focusing on useful science applied to, derived from, or inspired by living systems. The major themes that TIBTECH is interested in include: Bioprocessing (biochemical engineering, applied enzymology, industrial biotechnology, biofuels, metabolic engineering) Omics (genome editing, single-cell technologies, bioinformatics, synthetic biology) Materials and devices (bionanotechnology, biomaterials, diagnostics/imaging/detection, soft robotics, biosensors/bioelectronics) Therapeutics (biofabrication, stem cells, tissue engineering and regenerative medicine, antibodies and other protein drugs, drug delivery) Agroenvironment (environmental engineering, bioremediation, genetically modified crops, sustainable development).