Agrobacterium-Mediated Genetic Transformation in Wheat

Wheat (Triticum aestivum L.) is an important staple food crop that significantly impacts global food security. In terms of the need for an efficient genetic transformation protocol in major cereal crops, wheat comes immediately after rice to enhance their productivity level under varied global climatic conditions. Despite the availability of various Agrobacterium tumefaciens-based genetic transformation techniques for wheat crops, it remains the most challenging process and efficient transformation protocol continues to be a major hurdle in the development of genetically modified crops. Here in this study, we are describing the novel modified Agrobacterium tumefaciens-based in planta genetic transformation method which targets the 2-day-old apical meristematic germinated tissue as an explant for introgression of transgene. Germinated plants infected with Agrobacterium are screened for the presence of transgenes using PCR with gene-specific primers at the T₀ stage. The T₁ ds obtained from PCR-positive plants at T₀ generation are subjected to hygromycin B treatment. Surviving plants are then screened for transgene presence through PCR analysis, dot blot, and Southern hybridization. The expression of the transgene in the transgenic lines is assessed using qRT-PCR. The modified Agrobacterium-based in planta genetic transformation method for wheat is efficient, time-saving, and less labor-intensive. By introducing this method of plant transformation, the significant challenging obstacles in the genetic modification of wheat through genome editing or transgenic approaches to wheat are successfully addressed.