Abstract
The present work is an attempt to develop transgenic hairy root system in Camellia assamica, using leaves infected by Agrobacterium rhizogenes strain LBA9402, harboring binary vector (pART27 and pBI121) carrying β-glucuronidase (gus) as reporter gene and neomycin phosphotransferase (nptII) as selection marker. The transformed hairy roots were grown in phytohormone free MS media in the presence of kanamycin 50 µg/ml. The incorporation and expression of foreign genes were checked by initial gus assay followed by PCR analysis for the presence of nptII and β-glucuronidase (gus) gene. This protocol therefore facilitates the study of gene expression system in general and for root functional genomics in particular, giving scope for transgenic hairy roots as future explants for secondary metabolite production and plantlet regeneration. Key words: Agrobacterium rhizogenes, transformation, Camellia assamica, nptII, β-glucuronidase (gus).
Highlights
Tea is a widely consumed non-alcoholic and healthy beverage in the world belonging to the family of Theaceae
The present study focuses mainly the use of reporter gene in tea leaf explants that allows easy selection of transgenic lines through monitoring the function of reporter genes in the transgenic hairy roots developed through infection with A. rhizogenes strain LBA 9402
Transformation and selection of A. rhizogenes harboring the binary vector in tea
Summary
In many countries including India, tea is an important cash crop and has great value as a source of secondary metabolic products. The growing industrialization has always demanded for continual improvement and an increased productivity of this crop but the recalcitrant nature of this woody perennial plant has restricted its scope for genetic improvement unlike other woody perennials, breeding practices followed in tea takes a longer time because of long life cycle and selfincompatible nature of the crop (Banerjee, 1992). The effective strategy to overcome the problem could be the use of biotechnological approaches which lessens the constraints for genetic transformation and the time required to obtain a desired character. The natural responsiveness of plants towards transformation manifested the study for genetic analysis in perennial plant like tea.
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