Establishment of Suspension Cell Culture from &lt;em&gt;Agrobacterium&lt;/em&gt;-transformed Hairy Root Cells of &lt;em&gt;Psammosilene tunicoides&lt;/em&gt;, an Endangered and Rare Medicinal Plant of China

Zhang Zong-Shen,Wang Chao,Jin Zhao-Xia,Zhang Liang,Yu Zhen-Yan,Luo Huai-Yu,Pan Yan,Guo Qi-Gen

doi:10.19026/ajfst.9.1919

Abstract

Psammosilene tunicoides is an important medicinal plant endemic in China. Its annual yield is severely limited due to slow growth, poor seed germination and excessive collection. To satisfy the growing market demands, it’s necessary to seek alternatives to field cultivation and wild resources of this endangered plant. Using Agrobacterium -transformed hairy roots as initial explants, here, we reported the development of a suspension cell culture system for P. tunicoides . Results showed the Agrobacterium -transformed hairy roots-derived suspension cells are fast in growth and strong in capacity for accumulation of bioactive metabolites. We established that 1/2MS was a suitable medium for culturing the hairy root-derived suspension cells and the optimal combination of phytohormones is 1.5 mg/L 2, 4-D+0.5 mg/L 6-BA+0.25 mg/L NAA+0.1 mg/L KT. Under this condition, the maximal biomass was achieved at the 20 th day of culture with an average growth rate of 0.72 g/L/d; and the intracellular saponine content reached 0.92%, comparable to that of mother hairy roots. Compared with the normal P. tunicoides suspension cells, the hairy roots-derived suspension cells exhibited features of fast growth, short culture period and high concentration of saponines, suggesting that the large scale culture of hairy root-derived cells could be a feasible alternative to the wild resources of P. tunicoides .

Highlights

Comparing with the normal suspension cells, we showed that the hairy roots-derived cell culturing is a promising approach to produce suspension cells of P. tunicoides on industrial scale
Induction of P. tunicoides hairy roots: Leaves of sterilized seedlings were infected with Agrobacterium rhizogenes ACCC10060 for initiation of hairy roots and the hairy roots were cultured and maintained on solid MS supplemented with 2% sucrose + 0.45% agar as previously described (Li et al, 2011)
The results presented in this study clearly indicated that hairy roots-derived Suspension Cells (TSC) culture system exhibited remarkable advantages compared to suspension cells from normal explants (NSC) in the respects of growth rate and secondary metabolites accumulation

Summary

INTRODUCTION

Recent studies demonstrated the total of these components in this plant is much more effective, indicating that commercial supply of the P. tunicoides herbs heavily depends on over-collection of the wild plants (Zhang et al, 2012). Suspension culturing of cells established through callus which are induced from hairy roots will be a suitable alternative technology to overcome these problems, given its merits of good dispersion, stable genetics and fast growth rate of parental hairy roots. We have reported the success of induction and establishment of liquid culture system of P. tunicoides hairy roots (Li et al, 2011). Comparing with the normal suspension cells, we showed that the hairy roots-derived cell culturing is a promising approach to produce suspension cells of P. tunicoides on industrial scale

MATERIALS AND METHODS

AND DISCUSSION

CONCLUSION