Stable Maize Transformation Protocols

Biolistic Transformation

This detailed protocol by Manish Raizada describes stable transformation of maize (TypeII callus) using biolistic transformation.  It includes details on initiating maize callus from immature embryos, callus maintenance, transformation, Basta selection and regeneration.  Also included are media recipes:

1. Maize Transformation Protocol.pdf
2. Shooting maize callus.pdf
3A. Media - TypeII Callus Initiation.pdf
3B. Media - TypeII Callus Initiation.pdf
4. Transformation Media-osmotic.pdf
5. Media - Callus Selection.pdf
6. Media - Regen 1.pdf
7. Media - Regen 2.pdf
8. Media-stock solutions.pdf
9. Suppliers of Materials.pdf
10. Transformation References.pdf
11. Appendix - Light conversion.pdf
12. Transformation Contacts List.pdf

Agrobacterium Transformation
PDF of Agrobacterium transformation protocol from Kan Wang’s group at Iowa State University from Plant Physiology (2002)

The United States Patent Office also publishes very detailed protocols and information on patented maize transformation protocols (search under “maize AND transformation” in Abstracts):
http://www.uspto.gov/patft/

Patents of interest:

1. United States Patent  #6,822,144  Zhao , et al. Nov 23, 2004
Methods for Agrobacterium-mediated transformation

Abstract

The present invention relates to methods for improving the transformation frequency of Agrobacterium-mediated transformation of maize embryos. A preferred method for transforming maize using Agrobacterium comprises the steps of: contacting at least one immature embryo from a maize plant with Agrobacterium capable of transferring at least one gene to said embryo; co-cultivating the embryos with Agrobacterium; culturing the embryos in medium comprising N6 salts, an antibiotic capable of inhibiting the growth of Agrobacterium, and a selective agent to select for embryos expressing the gene; and regenerating plants expressing the gene.

2. United States Patent #6,403,865 Koziel , et al.  June 11, 2002
Method of producing transgenic maize using direct transformation of commercially important genotypes

Abstract

Methods for transformation of maize with nucleic acid sequences of interest are disclosed. The method involves subjecting immature zygotic embryos or Type I callus to high velocity microprojectile bombardment. The method is capable of producing transformed maize lines of commercial importance and their hybrid combinations.