Li Ying, Sun Min, Wang Xin, Zhang Yue-Jing, Da Xiao-Wei, Jia Ling-Yun, Pang Hai-Long, Feng Han-Qing
School of Life Sciences, Northwest Normal University, Lanzhou, 730070, China.
Bioresour Bioprocess. 2021 Dec 14;8(1):124. doi: 10.1186/s40643-021-00480-5.
In the last decades, replicating expression vectors based on plant geminivirus have been widely used for enhancing the efficiency of plant transient expression. By using the replicating expression vector derived from bean yellow dwarf virus and green fluorescent protein as a reporter, we investigated the effects of α-naphthalene acetic acid, gibberellins, and 6-benzyladenine, as three common plant growth regulators, on the plant biomass and efficiency of transient expression during the process of transient expression in Nicotiana benthamiana L. leaves.
With the increase of the concentration of α-naphthalene acetic acid, gibberellins, and 6-benzyladenine (from 0.1 to 1.6 mg/L), the fresh weight, dry weight, and leaf area of the seedlings increased first and then returned to the levels similar to the controls (without chemical treatment). The treatment with α-naphthalene acetic acid at 0.2 and 0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.4 mg/L α-naphthalene acetic acid and was increased about by 19%, compared to the controls. Gibberellins at 0.1-0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.2 mg/L gibberellins and was increased by 25%. However, the application of 6-benzyladenine led to decrease in the level of transient expression of green fluorescent protein.
The appropriate plant growth regulators at moderate concentration could be beneficial to the expression of foreign genes from the Agrobacterium-mediated transient expression system in plants. Thus, appropriate plant growth regulators could be considered as exogenous components that are applied for the production of recombinant protein by plant-based transient expression systems.
在过去几十年中，基于植物双生病毒的复制型表达载体已被广泛用于提高植物瞬时表达的效率。我们以菜豆黄矮病毒衍生的复制型表达载体和绿色荧光蛋白作为报告基因，研究了三种常见植物生长调节剂α-萘乙酸、赤霉素和6-苄基腺嘌呤对本氏烟草叶片瞬时表达过程中植物生物量和瞬时表达效率的影响。
随着α-萘乙酸、赤霉素和6-苄基腺嘌呤浓度的增加（从0.1至1.6毫克/升），幼苗的鲜重、干重和叶面积先增加，然后恢复到与对照（未进行化学处理）相似的水平。0.2和0.4毫克/升的α-萘乙酸处理可提高绿色荧光蛋白的瞬时表达水平，在0.4毫克/升α-萘乙酸时达到峰值，与对照相比增加了约19%。0.1至0.4毫克/升的赤霉素可提高绿色荧光蛋白的瞬时表达水平，在0.2毫克/升赤霉素时达到峰值，增加了25%。然而，6-苄基腺嘌呤的应用导致绿色荧光蛋白的瞬时表达水平降低。
适度浓度的合适植物生长调节剂可能有利于植物中农杆菌介导的瞬时表达系统中外源基因的表达。因此，合适的植物生长调节剂可被视为用于基于植物的瞬时表达系统生产重组蛋白的外源成分。
