A simple and efficient protocol for hairy root culture of Arabidopsis thaliana

Hairy root culture (HRC) represent a valuable biotechnological tool for the production of plant secondary metabolites. Secondary metabolome study of Arabidopsis thaliana may help to understand the biological roles of various secondary metabolites present in it. The present work deals with the establishment of Agrobacterium rhizogenes strains transformed HRC of A. thaliana with very high transformation frequency resulting in long term hairy root cultures grown in hormone free media. Optimization of culture medium and standardisation of co-cultivation period are the key role players in obtaining high frequency of hairy roots. Four days of preculture in CIM medium and five min of co-cultivation in the bacterial suspension were found to be optimal conditions for root induction. This protocol could become a powerful tool for transcriptomics, metabolomics and proteomics-based studies for different transgenic root lines of A. thaliana. Insertion of rol genes of Agrobacterium rhizogenes enhanced rhizogenic potential of excised explants in Arabidopsis thaliana. The optimization process of hairy root culture induction and establishment led to maintenance upto 4 years in hormone free media.