Transformation improvement with the Standardized Pressure Agrobacterium Infiltration Device (SPAID)

The treatment of plant tissue with Agrobacterium tumefaciens is often a critical first step to both stable and transient plant transformation. In both applications bacterial suspensions are oftentimes physically introduced into plant tissues using hand-driven pressure from a needleless syringe. Whil...

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Bibliographic Details
Published in:	BMC research notes Vol. 12; no. 1; p. 144
Main Authors:	Mad' Atari, Mohamad Fadhli Bin, Folta, Kevin M
Format:	Journal Article
Language:	English
Published:	England BioMed Central Ltd 15-03-2019 BioMed Central BMC
Subjects:	Adventitious shoots Agrobacterium Agrobacterium tumefaciens - genetics Arabidopsis Arabidopsis - genetics Arabidopsis thaliana Bacteria Contact angle Containers Diploid strawberry Ethanol Fragaria - genetics Gene expression Genetic Engineering - instrumentation Genetic Engineering - methods Laboratories Localization Methods Nicotiana - genetics Plant Leaves - genetics Plants, Genetically Modified Pressure Proteins Reproducibility of Results Research Note Stable Strawberries Transformation Transformation, Genetic Transient Arabidopsis thaliana Nicotiana benthamiana Transformation Agrobacterium Stable Transient Adventitious shoots Fragaria vesca Diploid strawberry
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Summary:	The treatment of plant tissue with Agrobacterium tumefaciens is often a critical first step to both stable and transient plant transformation. In both applications bacterial suspensions are oftentimes physically introduced into plant tissues using hand-driven pressure from a needleless syringe. While effective, this approach has several drawbacks that limit reproducibility. Pressure must be provided with the syringe perfectly perpendicular to the tissue surface. The researcher must also attempt to provide even and consistent pressure, both within and between experimental replicates. These factors mean that the procedures do not always translate well between research groups or biological replicates. We have devised a method to introduce Agrobacterium suspensions into plant leaves with greater reproducibility. Using a decommissioned dissecting microscope as an armature, a syringe body with the bacterial suspension is mounted to the nosepiece. Gentle, even pressure is applied by rotating the focus knob. The treatment force is measured using a basic kitchen scale. The development of the Standardized Pressure Agrobacterium Infiltration Device (SPAID) provides a means to deliver consistent amounts of bacterial suspensions into plant tissues with the goal of increasing reproducibility between replicates and laboratories.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1756-0500 1756-0500
DOI:	10.1186/s13104-019-4117-3