Transformation of
View Patent ↗The present invention relates to a method for transforming Allium species with a heterologous gene using Agrobacterium.
1. A method for transforming a plant or plant tissue of an Allium cepa or Allium fistulosum with a DNA of interest from a heterologous gene, the method comprising the steps of: contacting an embryogenic callus from a plant of an Allium cepa or Allium fistulosum with a bacterium belonging to the genus Agrobacterium which contains a DNA of interest from a heterologous gene and obtaining a transformed Allium cepa or Allium fistulosum embryogenic callus under selective conditions.
2. The method of claim 1 wherein the bacterium belonging to the genus Agrobacterium is Agrobacterium rhizogenes or Agrobacterium tumefaciens.
3. The method of claim 1 wherein the bacterium belonging to the genus Agrobacterium contains a Ti plasmid or a Ri plasmid.
4. The method of claim 1 wherein the heterologous gene is the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene.
5. The method of claim 4 wherein the heterologous gene is a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene which encodes an enzyme that confers resistance to the herbicide glyphosate.
6. The method of claim 1 wherein the embryogenic callus is derived from immature embryos or flower buds from an Allium cepa or Allium fistulosum.
7. An Allium plant or plant tissue transformed by the method of claim 1 or transformed progeny thereof each containing a DNA of interest comprising a 5-enolpvruvylshikimate-3-phosphate synthase (EPSPS) gene.
8. A method for transforming a plant or plant tissue of an Allium cepa or Allium fistulosum with a DNA of interest from a heterologous gene, the method comprising the steps of:
a. culturing immature embryos or flower buds from a plant of an Allium cepa or Allium fistulosum on an initiation medium for a period of from 2 to 6 months until an embryogenic callus forms on the embryos or flower buds:
b. transferring the embryonic callus to a coculture medium and contacting the embryogenic callus with a suspension of Agrobacterium rhizogenes or Agrobacterium tumefaciens containing a DNA of interest from a heterologous gene; and
c. obtaining a transformed Allium cepa or Allium fistulosum embryogenic callus under selective conditions.
9. The method of claim 8 wherein the immature embryos or flower buds are cultured on the initiation medium in the dark and at a temperature of from 25° C. to 30° C.
10. The method of claim 8 wherein the heterologous gene is the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene.
11. The method of claim 10 wherein the heterologous gene is a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene which encodes an enzyme that confers resistance to the herbicide glyphosate.
12. The method of claim 8 further comprising the step of regenerating the transformed embryonic callus into transformed Allium plants containing the DNA of interest from the heterologous gene.
13. An Allium plant or plant tissue transformed by the method of claim 8 or transformed progeny thereof each containing a DNA of interest comprising a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene.