Stage I – Making Hairy Roots








Target Plant
Seeds
Plantlet
Infect Plantlet
Initial Roots/Mother Culture
Natural
A. rhizogenes
Bacterial
Culture
A. rhizogenes with
recombinant (R) gene*







Establishing a hairy root (HR) culture begins with the selection of a target plant, from which viable seeds are obtained. Seeds are germinated and the resulting plantlets are infected either with a bacterial suspension of a naturally occurring strain (wild type) of Agrobacterium rhizogenes, or a version that has been modified to contain a recombinant gene (R) encoding a desired protein. Infected plantlets are transferred to tubes containing solid agar until the appearance of nascent hairy roots. This constitutes a mother HR culture.
Stage II – Propagating and Processing Hairy Roots
and Alternative Plant Cultures









Culture HR
Suspension
Final
Concentrate
Micropropagation
Callus Cultures
Callus
Induction
Transfer
Expand HR
Biomass
Plant
Hormones
Extract
Plant
Extract
Harvest HR Tissue
&
Culture Media
Proprietary, Scalable,
Modular Bioreactors
Recombinant
Proteins
High-value
Phytochemicals












The HR tissue from mother cultures is excised and further grown in plastic dishes until attaining sufficient size to be transferred to flasks with liquid culture media where they are propagated in suspension. Once they reach a critical size, the HR cultures can be transferred to proprietary scalable, modular bioreactors to produce large quantities of HR biomass. Alternatively, suspension HR cultures can be placed under special conditions to promote the formation of callus aggregates, which in turn can be used to generate many identical plants through micropropagation in the presence of appropriate plant hormones and growth factors. The resulting plant tissue (either HR biomass or micropropagated plants), as well as the conditioned culture media, are harvested and used to make extracts enriched in high-value phytochemicals (liquid or powder) or a recombinant protein of interest if the initial infection was performed with a strain of A. rhizogenes modified to include a recombinant gene as shown in Stage I.