ArticlePlant cell reports2009
Optimization of conditions for transient Agrobacterium-mediated gene expression assays in Arabidopsis.
Article in Plant cell reports, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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47 citing papers in PubMed.
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- A high-efficiency transient expression system mediated by Agrobacterium tumefaciens in Spinacia oleracea leaves.Plant methods · 2024Article
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- Crop genome editing through tissue-culture-independent transformation methods.Frontiers in genome editing · 2024Review
- Identification of a suitable method of inoculation for reducing background effect in mock-inoculated controls during gene expression studies inPhysiology and molecular biology of plants : an international journal of functional plant biology · 2023Article
- New methods for sorghum transformation in temperate climates.AoB PLANTS · 2023Review
- Effects of different light conditions on transient expression and biomass inOpen life sciences · 2023Article
- Evaluation of novel surfactants for plant transformation.BMC research notes · 2022Article
- Plant Molecular Pharming and Plant-Derived Compounds towards Generation of Vaccines and Therapeutics against Coronaviruses.Vaccines · 2022Review
- The Characterization of theInternational journal of molecular sciences · 2022Article
- An efficientFrontiers in plant science · 2022Article
- An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber.Frontiers in plant science · 2022Article
- A high-efficiency Agrobacterium-mediated transient expression system in the leaves of Artemisia annua L.Plant methods · 2021Article
- Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants.International journal of molecular sciences · 2021Review
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3 authors.
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Abstract
Transient genetic transformation of plant organs is an indispensable way of studying gene function in plants. This study was aimed to develop an optimized system for transient Agrobacterium-mediated transformation of the Arabidopsis leaves. The beta-glucuronidase (GUS) reporter gene was employed to evaluate growth and biochemical parameters that influence the levels of transient expression. The effects of plant culture conditions, Agrobacterial genetic backgrounds, densities of Agrobacterial cell suspensions, and of several detergents were analyzed. We found that optimization of plant culture conditions is the most critical factor among the parameters analyzed. Higher levels of transient expression were observed in plants grown under short day conditions (SDs) than in plants grown under long day conditions (LDs). Furthermore, incubation of the plants under SDs at high relative humidity (85-90%) for 24 h after infiltration greatly improved the levels of transient expression. Under the optimized culture conditions, expression of the reporter gene reached the peak 3 days after infiltration and was rapidly decreased after the peak. Among the five Agrobacterial strains examined, LAB4404 produced the highest levels of expression. We also examined the effects of detergents, including Triton X-100, Tween-20, and Silwet L-77. Supplementation of the infiltration media either with 0.01% Triton X-100 or 0.01% Tween-20 improved the levels of expression by approximately 1.6-fold. Our observations indicate that transient transformation of the Arabidopsis leaves in the infiltration media supplemented with 0.01% Triton X-100 and incubation of the infiltrated plants under SDs at high relative humidity are necessary for maximal levels of expression.
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