Agrobacterium rhizogenes-mediated hairy roots transformation as a tool for exploring aluminum-responsive genes function

Abstract

Aim: To develop a useful alternative approach to evaluate the gene function in hairy roots. Methods: Arabidopsis and tobacco (wild-type or mutant) were a host for Agrobacterium rhizogenes transformation. Results: The hairy roots formation efficiency ranged from 53 to 98% in tobacco and 53 to 66% in Arabidopsis. Hairy and intact roots showed similar gene expression pattern in response to salt and aluminum stress. Genomic polymerase chain reaction and fluorescent images showed high rate (>80%) of co-integration of T-DNAs and uniform cell transformation without use of any antibiotic selection. Whole processes of hairy roots were completed within 1 month after the infection of Agrobacterium. Conclusion: Aluminum-responsive orthologous gene function could be evaluated by NtSTOP1-KD and Atstop1 as a host for hairy roots transformation. Hairy roots have been used for various purposes over the last several years. In the present study we used Agrobacterium rhizogenes-mediated hairy roots as an alternative approach for Agrobacterium tumefaciens transformation. We developed a simple, effective and reproducible hairy root protocol in tobacco and Arabidopsis. Developed hairy roots were compared with intact roots to characterize the gene response to aluminum, NaCl stress and cellular localization of genes. Aluminum-responsive orthologous genes function could be evaluated by using NtSTOP1-KD and Atstop1 as a host for hairy roots transformation.