Method for boosting genetic transformation efficiency of wheat using agrobacterium-mediated method
Disclosed is a method for boosting genetic transformation efficiency of wheat using an Agrobacterium -mediated method, which belongs to the field of plant genetic engineering technology. A TaHRF2 gene and an encoded protein thereof used in the present disclosure can promote introduction of a nucleic acid molecule into a plant of interest and boost genetic transformation efficiency of the introduction of the nucleic acid molecule into the plant of interest. The plant of interest includes, but is not limited to, monocotyledonous plants such as wheat. By introducing a TaHRF2 gene high-expression vector into an Agrobacterium strain, and infecting a wheat explant using an Agrobacterium -mediated method, genetic transformation capacities of backbone parents such as Beijing 8 and Lumai 1 are enhanced, providing important information and fundamental technical support for the establishment of an efficient genetic transformation system for major wheat cultivars with difficulty in regeneration.
1 . A method for boosting transformation efficiency of wheat using Agrobacterium -mediated transformation, the method comprising:
ligating a TaHRF2 gene into an expression vector to construct a recombinant expression vector, and transforming the recombinant expression vector into an Agrobacterium competent cell to obtain an Agrobacterium strain for use in transformation; and
infecting a wheat immature embryo with the Agrobacterium strain, thereby boosting transformation efficiency and obtaining a wheat transgenic and gene-edited plant;
wherein the TaHRF2 gene is a DNA molecule selected from the group consisting of i) and ii) below:
i) the DNA molecule having the nucleotide sequence set forth in SEQ ID NO. 1; and
ii) the DNA molecule encoding the amino acid sequence set forth in SEQ ID NO. 2;
wherein the wheat is of variety “Beijing 8” or “Lumai 1”.
2 . The method of claim 1 , wherein the expression vector is a pUbi110 plasmid.
3 . The method of claim 1 , wherein the Agrobacteriu m competent cell is Agrobacteriu m EHA105.
4 . The method of claim 1 , wherein a protein encoded by the TaHRF2 gene is a protein selected from the group consisting of (A1) and (A2) below:
(A1) the protein consisting of the amino acid sequence set forth in SEQ ID NO. 2 of the Sequence Listing; and
(A2) a fusion protein obtained by attaching a protein tag to the N-terminus and/or the C-terminus of the protein defined in (A1).