摘要:Abstract•This work describes a protocol for hairy root transformation of the medicinal crop legume fenugreek (Trigonella foenum-graecum L.). Hairy root plant transformation mediated byAgrobacterium rhizogenesis an established method for the rapid genetic transformation of various dicotyledonous plants. We have adapted a hairy root transformation protocol from the model legumeMedicago truncatulafor use in this metabolically rich non-model crop legume. Considering the great variety and abundance of phytochemicals in fenugreek and its established use in traditional medicine, we aim for this method to become a resource for metabolic pathway identification and for production of valuable specialised metabolites via metabolic engineering approaches.•Development rapid transformation (2.5–3 weeks) of fenugreek roots viaA. rhizogenes.•Marker gene cassette with suitable promoter for visual detection of transformed fenugreek rootsGraphical abstractDisplay Omitted