Production of Doubled Haploid Transgenic Wheat (Triticum aestivum L.): Transformation of microspores and regeneration of homozygous transformants

Production of transgenic wheat with desired traits is of great importance. The efficiency in obtaining transgenic wheat plants is low and labor intensive, and it may take years to obtain the desired homozygous transgenic plant in the desired genotypic background. This work describes a microspore transformation method for the production of large number of homozygous doubled haploid transgenic wheat in one generation. In the first part of this work, optimal conditions for generating large numbers of DH wheat plants from microspores were identified. In the second part of this work, microspores were transformed by co-cultivation with Agrobacterium tumefaciens. PCR, DNA sequencing, Southern blot analyses and assay of the recombinant enzyme confirmed the presence of transgenes in T0 primary transformants and their stable inheritance in homozygous T1 DH progenies. Several factors for successful transformation were identified. The method described and conditions worked out in this work are likely to be applicable to other plant species, and should be especially useful to professionals in universities, research institutions and agricultural industry.