Comprehensive Optimization of Agroinfiltration in Transient Expression System in Nicotiana tabacum with RUBY Visible Reporter.

Q3 Agricultural and Biological Sciences

Doklady Biological Sciences Pub Date : 2026-02-01 Epub Date: 2026-03-30 DOI:10.1134/S0012496625600721

M R Sorokina, Yu A Yugay, A I Degtyarenko, A A Sadykov, Yu N Shkryl, V P Bulgakov

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引用次数: 0

Abstract

Agrobacterium-mediated transient expression is one of the most popular tools in functional genomics and plant biotechnology. However, its efficiency and reproducibility depend on multiple factors. Agroinfiltration conditions were optimized comprehensively to enhance the expression level of betalain biosynthetic genes in the model plant Nicotiana tabacum with the use of the RUBY visual reporter system. The effects of bacterial suspension concentration, infiltration buffer composition, and delivery method were evaluated. The highest expression efficiency was achieved with a suspension of OD₆₀₀ = 1.0; a buffer containing acetosyringone (50 μM), 2-(N-morpholino)ethanesulfonic acid (MES) (10 mM), sucrose (0.5%), and magnesium chloride (10 mM); and delivery via preliminary leaf micropuncture with a needle roller and subsequent vacuum infiltration. The resulting protocol significantly improves the efficiency of transient expression and can be employed in gene functional analysis, metabolic engineering, and production of biopharmaceutical compounds in plants.

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利用RUBY可视化报告软件对烟草瞬态表达体系中土壤渗透的综合优化。

农杆菌介导的瞬时表达是功能基因组学和植物生物技术中最常用的工具之一。然而，其效率和重现性取决于多种因素。利用RUBY可视化报告系统，综合优化农田入渗条件，提高模式植物烟叶中甜菜素生物合成基因的表达水平。评价了细菌悬浮液浓度、浸润缓冲液组成和递送方式的影响。OD600 = 1.0的悬液表达效率最高；缓冲液含有乙酰丁香酮（50 μM）、2-(N-morpholino)乙磺酸（MES）（10 mM）、蔗糖（0.5%）和氯化镁（10 mM）；用滚针器对叶片进行初步微穿刺然后进行真空渗透。该方案显著提高了瞬时表达效率，可用于植物基因功能分析、代谢工程和生物制药化合物的生产。

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

Doklady Biological Sciences Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)

CiteScore

1.10

自引率

0.00%

发文量

期刊介绍： Doklady Biological Sciences is a journal that publishes new research in biological sciences of great significance. Initially the journal was a forum of the Russian Academy of Science and published only best contributions from Russia in the form of short articles. Now the journal welcomes submissions from any country in the English or Russian language. Every manuscript must be recommended by Russian or foreign members of the Russian Academy of Sciences.