PJB-2022-522
Tissue Culture Optimization and Genetic Transformation of Fragaria Vesca L. to Enhance its Antioxidant Potential
Afshan Anwar
Abstract
Tissue Culture Optimization and Genetic Transformation of Fragaria Vesca L. to Enhance its Antioxidant Potential
Afshan Anwar1, Shabana Haneef 1,2*, Khadija Tahir1, Iram Murtaza1, Bushra Mirza1
1 Department of Biochemistry, Faculty of Bioscience, Quaid I Azam University, Islamabad, Pakistan
2 Alpha Genomics Private Limited, PWD, Islamabad, Pakistan
*Correspondence: shabana.hanif@bs.qau.edu.pk
Abstract: Fragaria vesca L. is, prototypical for molecular and genetic research, the representative of the Rosaceae family. Being ideal it has a small life cycle, facile seed germination and vegetative propagation along with a small diploid genome that is already sequenced. Plants secondary metabolites productivity can be enhanced by using rol genes of Agrobacterium rhizogenes, however, rolB is the most influential stimulator. We aimed to elucidate the rolB functions in this model plants by designing an effectual, reproducible Agrobacterium-mediated prototype for its genetic transformation. Tissue culturing medium along with indole-3-butyric acid and benzyladenine combination was optimized for transgenic plants regeneration of shoots. Agrobacterium tumefaciens strain LBA4404, harbouring binary vector pLBR30 having rolB expression cassette used. Transgenic were obtained by determining the concentrations of antibiotics that have the least phytotoxic effects on tissue regeneration and are effective in the elimination of Agrobacterium and selection of transformant of rolB respectively. Molecular and expressional analysis was confirmed for the successful transgenic integration. Further antioxidant assays were performed to determine the effect of transgene on the antioxidant potential of F. vesca. Transgenic lines showed increased antioxidant activity correlated with increased secondary metabolites as compared to wild-type plants. Phenotypic alterations in growth associated with rolB gene were also observed in all transgenic plants. Concluding, this prototype can be used for the genetic transformation of plants with rolB gene to yield transgenic fruits, crop species with increased antioxidant potential.
Keywords: Fragaria vesca L., Agrobacterium-mediated transformation, rolB gene, Antioxidant assays
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