Optimizing hairy root production from explants of Phyllanthus hainanensis, a shrub used for traditional herbal medicine

Abstract

Phyllanthus hainanensis is a shrub that has been used in traditional herbal medicine. It has great pharmaceutical potential for treating diseases such as cancer and diabetes. As a prerequisite for propagation of this species on a large scale, hairy roots in P. hainanensis were induced using Rhizobium rhizogenes and various factors affecting hairy root induction and growth evaluated. Seven factors were tested: (1) type of explant, (2) type of culture medium, (3) duration of pre-culture, (4) R. rhizogenes inoculum cell density, (5) duration of infection, (6) acetosyringone concentration in the culture medium, and (7) duration of incubation. The optimal protocol for hairy root induction and growth was: young shoots, pre-cultured in Y1 for 2 d, inoculated with R. rhizogenes broth with an OD₆₀₀ of 0.6 for 20 min, and incubated for 3 d. Putative transgenic hairy roots were initially identified by morphology and then confirmed by polymerase chain reaction. Successful and optimal production of hairy roots is a critical prerequisite for industrial scale clonal propagation of P. hainanensis. Being able to cultivate the plant on a large scale will provide rapid and ready supply of the plant materials that can be used in herbal medicine and in scientific and industrial exploitation.

Keywords

hairy root induction / medicinal plant / MS growth medium / Phyllanthus hainanensis / Rhizobium rhizogenes

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Acknowledgements

This work was supported by China Research Funds for Universities (2662016QD021).

Compliance with ethics guidelines

Zhaogui Yan, Shenyu Liu, Junlian Zhang, Guan Huang, Lijun Duan, and Yaomei Ye declare that they have no conflicts of interest or financial conflicts to disclose.

This article does not contain any studies with human or animal subjects performed by any of the authors.

RIGHTS & PERMISSIONS

The Author(s) 2019. Published by Higher Education Press. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0)