Vol. 9, Issue 6, Part K (2024)

Author(s): Bhoomikaben A Joshi, Chintan Kapadia and Alok Vaidya

Abstract: The post-harvest losses are mainly due to over-ripening of fruits or vegetables. The main hormone responsible for fruit ripening is ethylene, which is also produced in excess under stress conditions. ACC (1-aminocyclopropane-1-carboxylic acid) is a precursor of ethylene. Enzyme ACC deaminase breaks ACC and thus helps reduce ethylene levels in plants. Pectin is a major part of the plant cell wall, and enzyme pectin methylesterase (PME) breaks the pectin, ultimately loosening the cell wall, which greatly affects physiochemical plant activities. Enzyme pectin methylesterase inhibitor (PMEI) inhibits PME and reduces cell wall breakage. Both genes were isolated, fused together, and transformed into Pichia pastoris. Gene ACC deaminase was isolated from the fungus Trichoderma asperellum, and gene PMEI was isolated from Solanum lycopersicum (tomato), which were 1118 bp and 555 bp in size, respectively. Both genes were linked with YFP and GFP and cloned into the pET101 vector, then fused together using a 2A peptide linker. The fused construct was cloned into the pPIC9 vector and transformed into Pichia pastoris. From transformed cells, single-colony PCR results showed the amplification of both genes. The biochemical tests of both genes further demonstrated protein expression in the host. This study ultimately helps in increasing fruit shelf life and reducing post-harvest losses by mitigating abiotic stress.

DOI: 10.22271/veterinary.2024.v9.i6k.1972