Enhanced Antioxidant and Anticancer Potential of Buch Transformed with Gene.

Secondary metabolites have been shown to possess a range of biological functions. Flavonoids, due to their ability to scavenge ROS, are famous antioxidants. The plants of species are rich sources of flavonoids; however, the amount of these metabolites is less. In the current study, the flavonoid content was detected and then enhanced by genetically modifying the Buch with strain GV3101 carrying gene. The transformation of gene was confirmed with PCR and the gene copy number was confirmed by Southern blot analysis. The HPLC analysis revealed the presence of catechin (3.19 ug/mg DW) and geutisic acid (2.22 ug/mg DW) in transformed plants, unlike wild-type plants. In transformed plants, all detected flavonoids (vanillic acid, rutin, catechine, gallic acid, syringic acid, caffeic acid, coumaric acid, geutisic acid, ferulic acid, and cinnamic acid) were increased up to several folds. Real-time qPCR revealed the higher expression levels of the genes for flavonoid biosynthesis enzymes phenylalanine ammonia-lyase and chalcone synthase in plants transformed with genes, as the expression levels were increased up to 9-20-fold and 2-6-fold, respectively. The transgenic lines T3 and T5 carrying two copies of gene, particularly showed higher expression of both and gene, with the highest expression in T3 line. The transgenic lines demonstrated an average increase of 1.4-fold in the total phenolic content and 1-2-fold in the total flavonoid content as compared to wild-type plants. Total antioxidant capacity and total reducing power were increased up to an average of 1-2-fold and 1.5-2-fold respectively, along with increased free radical scavenging ability. Furthermore, the gene transgenics were found to have much greater cytotoxic capacity than the wild-type plant against the MCF7, HeLA, and HePG2 cancer cell lines. Current findings show that the gene effectively increases the flavonoid content of Buch, boosting the plant's capacity as an antioxidant and an anticancer. This is the first-ever report, demonstrating the genetic transformation of Buch with gene.