Precise plant breeding using plant DNA border like sequences instead of Agrobacterium borders

摘要

Disclosed is a method of modifying a trait of a selected plant by stably integrating only a desired polynucleotide into the genome of the plant, comprising the steps of: (a) transforming the selected plant with the desired polynucleotide, wherein: (i) the desired polynucleotide consists essentially of a nucleic acid sequence that is native to the selected plant, native to a plant from the same species as the selected plant, or native to a plant that is sexually interfertile with said selected plant, and wherein the desired polynucleotide is flanked by at least one border-like sequence that comprises the sequence SEQ ID NO: 93; (ii) the desired polynucleotide does not contain DNA that is not from: (1) the selected plant species or (2) a plant that is sexually compatible with the selected plant species; and (iii) the at least one border-like sequence does not comprise a nucleotide sequence that is identical in nucleotide sequence to an Agrobacterium TDNA border sequence; (b) selecting one or more plant cells from the plant transformed by step (a) that comprise only the desired polynucleotide in their genome; and (c) obtaining at least one stably transformed plant from the one or more selected transformed plant cells, wherein the transformed plant contains only the desired polynucleotide stably integrated into its genome and wherein the stably transformed plant exhibits a trait that is modified in comparison to the selected plant.