Gene cloning: Applications in vegetable crops

Abstract

Gene cloning is a revolutionary technique in agricultural biotechnology, particularly in vegetable farming, enabling precise genetic modifications to enhance crop traits such as pest resistance, disease tolerance, and nutritional improvement. The process involves inserting DNA fragments into vectors, which replicate within bacterial hosts. This facilitates large-scale gene duplication. Unlike PCR, gene cloning allows for the replication of longer DNA sequences, making it essential for studying gene functions. The first step involves isolating a target gene, typically from a cDNA library, to overcome the challenges of large plant genomes and introns. The Ti plasmid from Agrobacterium tumefaciens is widely used in plant genetic engineering, though modified binary vector systems improve its efficiency. Essential enzymes like restriction endonucleases, DNA ligases and polymerases facilitate DNA manipulation. Escherichia coli is the preferred host for cloning due to its ease of cultivation and rapid growth, while yeast and plant cell cultures serve as alternatives for eukaryotic gene expression. Different transformation methods are being used to introduce recombinant DNA into host cells. Successful transformants are identified through antibiotic selection, blue-white screening, PCR and DNA sequencing. Gene cloning has significant applications in vegetable science, enabling the development of stress-resistant, nutritionally enhanced crops, thereby contributing to sustainable agriculture and food security.