An Optimized Tobacco Hairy Root Induction System for Functional Analysis of Nicotine Biosynthesis-Related Genes

Rhizobium rhizogenes-mediated plant hairy root induction is a convenient method for functional study of root-specific genes. To develop an optimized tobacco system and gene function in nicotine biosynthesis, we investigated by three R. rhizogenes strains, R1601, K599, LBA9402, on different media with leaf discs from plants ages, observed that the strain LBA9402 used explant infection exhibited highest rate 4 8 week old ‘Coker176’ 2/3MS medium, it could also be as cargo delivering foreign genes to roots. Overexpression MsSPL12 gene, alfalfa (Medicago sativa) SQUAMOSA promoter binding protein-like (SPL) transcription factor, significantly improved production transgenic roots, reaching 1.38–1.85 mg/g compared 0.5 controls. Expression analysis biosynthesis transport-related responding methyl-jasmonate (MeJA) treatment revealed significant upregulation NtMPO2 responsible increased Our results establish high-throughput approach characterization roots elite cultivar, ‘Coker176’, well suggest efficiently manipulating biosynthesis.