Optimizing hairy root production from explants of Phyllanthus hainanensis,a shrub used for tradition

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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 Yl 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 pro-duction 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.
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