Rapid in-planta chemical method for plant genetic transformation without tissue culture

Overview

This solution offers a rapid and simple in-planta method for plant genetic transformation and genome editing that bypasses the need for traditional plant tissue culture. By applying a chemical agent mixed with a plasmid construct directly onto the emerging flower bud tissue of actively growing plants, the cell wall is disrupted, allowing recombinant DNA or guide RNA to enter the cell and integrate into the host genome. This approach addresses a major bottleneck in plant biotechnology, particularly for monocots, legumes, and economically important horticulture crops that are otherwise recalcitrant to standard transformation methods. The method has been validated on multiple model and crop species, including mungbean, chilli, papaya, and mango, and is under patent consideration in India.

Technical specifications

• Chemical-mediated delivery: A plant-compatible chemical disrupts the cell wall, enabling plasmid entry into meristematic cells without laboratory-based tissue culture steps.
• Direct flower bud application: The plasmid construct carrying a reporter gene cassette or guide RNA is mixed with the chemical at standardized concentrations and gently placed on the meristematic region of actively growing plants.
• Stable integration demonstrated: Transgene integration has been confirmed in leaves, pollen, and ovary tissue of putatively transformed T0 plants.
• Molecular validation: PCR analysis using gene-specific primers produced amplicons of expected size, confirming positive correlation with observed transformation results.
• Broad applicability: Suitable for food crops (monocots, legumes) and horticulture crops (chilli, papaya, mango) that are typically difficult to transform.
• Reduced timeline: Enables transformation during the window between flower initiation and fruit maturity, with transformed flower buds used for pollination to shorten the path to stable transformants.

Technology readiness level

The technology has been demonstrated and validated on multiple crop species including mungbean, chilli, papaya, and mango, with confirmed stable integration of transgenes in T0 plants verified through molecular analysis. The protocol is under consideration for patent protection in India. Future validation efforts will focus on standardizing the transformation method for flower buds across additional crops of interest, screening for gene inserts, pollination of transformed buds, seed harvesting, and screening for stable transformants in subsequent generations. Molecular analysis and field studies comparing transformed and control plants for agronomic performance are also planned to advance the technology toward broader deployment.