Protocol for Agrobacterium-mediated transformation in rice. Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2–3 months with a transformation frequency of 30–50%, both in easily cultured varieties and recalcitrant elite japonica rice. - [Read Agrobacterium-Mediated Transformation in Rice Protocol]
Electrotransformation of Agrobacterium with a plasmid that has been replicating in E. coli. Growth of Arabidopsis thaliana. Arabidopsis dunking. Seed Harvesting. Plant tissue culture. Very detailed protocol. Stockinger lab. PDF - [Read Arabidopsis transformation with Agrobacterium PDF]
Arabidopsis transformation and selection on kanamycin agar, Preparation of plants for transformation, Agrobacterium, Dipping, Post-dip care, Agar plates, Tips. Adaptation of the short protocol described bySteve Clough and Andrew Bent, University of Illinois at Urbana-Champaign. - [Read Arabidopsis transformation with pictures]
Virus-induced gene silencing (VIGS) uses a virus to deliver a sequence from a gene of interest into a host plant. The virus carrying the fragment of the gene of interest must be capable of replication if dsRNA is to be produced. One or two leaves are inoculated with Agrobacterium strains carrying the VIGS vector possessing the gene fragment. The virus then replicates and spreads throughout the plant, mediating silencing. - [Read Delivery of dsRNA into Plants by VIGS Methodology]
Protocol describes a method to determine the presence of plasmid DNA in an Agrobacterium culture. Compared to selection of transformed Agrobacterium, which can be ambiguous and normally takes several days for resistant colonies to appear, the approach described here is both rapid and accurate. - [Read PCR Analysis of Agrobacterium Protocol]
Arabidopsis can be stably transformed using Agrobacterium tumefaciens-mediated transfer of T-DNA. We describe the generation of transgenic plants via root transformation in tissue culture, which can be useful for transforming sterile mutants. - [Read Root Transformation of Arabidopsis Protocol]
Protocol describes a method for transforming Agrobacterium with plasmid DNA using electroporation in a manner similar to that commonly used for Escherichia coli. Although the transformation efficiency for Agrobacterium is lower than that for E. coli, it is possible to obtain adequate numbers of Agrobacterium transformants with this technique. - [Read Transformation of Agrobacterium Using Electroporation Protocol]
Protocol describes a method for transforming Agrobacterium with plasmid DNA using a freeze-thaw technique. Although the transformation efficiency for Agrobacterium is lower than that for Escherichia coli, it is possible to obtain adequate numbers of transformants with this technique. - [Read Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol]
The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]
A. tumefaciens is a soil-dwelling bacterium that transforms normal plant cells into tumor-forming cells by inserting a piece of bacterial DNA (the transfer, or "T," DNA) into the plant cell genome. The Ti plasmid also carries many of the transfer functions for mobilizing the T-DNA. This article provides a brief discussion of the principles of T-DNA transformation, including consideration of T-DNA vectors and their hosts. - [Read Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol]