The establishment of cell suspension culture of sabah snake grass (Clinacanthus nutans (Burm.F.) Lindau)

  • Qian Yi Phua
  • Sreeramanan Subramaniam
  • Vuanghao Lim
  • Bee Lynn Chew
Plant Tissue Culture


Clinacanthus nutans (Burm.F.) Lindau is an herbaceous plant that has long been used for traditional medicinal purposes in Asia. It has recently gained popularity as an alternative treatment for cancer. The aim of this study was to establish cell suspension cultures of C. nutans and to identify targeted bioactive compounds in the cultures. Young leaf explants were cultured on Murashige and Skoog medium supplemented with various combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin to identify a suitable medium for callus induction and proliferation. Proliferated, friable calluses were cultured in different combinations of plant growth regulators (2,4-D, naphthaleneacetic acid [NAA], picloram, kinetin, and 6-benzylaminopurine) in liquid medium to establish cell suspension cultures. Three cell lines of suspension culture, callus, and intact plant parts were subjected to ethyl acetate extraction followed by thin layer chromatography for identification of selected bioactive compounds. Medium supplemented with 0.25 mg L−1 2,4-D and 0.75 mg L−1 kinetin was found to be optimal for callus induction, whereas supplementation with 0.50 mg L−1 2,4-D was efficient for callus proliferation. Liquid medium supplemented with 0.25 mg L−1 2,4-D and 0.50 mg L−1 NAA produced the highest growth index (2.52). Quercetin, catechin, and luteolin were present together in the callus and cell suspension cultures of C. nutans, but all three compounds were detected separately in young leaves, mature leaves, and stems. This study is the first to report the establishment of cell suspension culture of C. nutans with both cell and callus cultures producing quercetin, catechin, and luteolin.


Callus induction Callus proliferation Cell suspension culture Thin layer chromatography Clinacanthus nutans 


Funding Information

The authors would like to acknowledge the Malaysian Ministry of Higher Education for funding the project under the Fundamental Research Grant Scheme (Grant code: 203/PBIOLOGI/6711521). They also thank Universiti Sains Malaysia and the Agricultural Crop Trust (Malaysia).


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Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  1. 1.School of Biological SciencesUniversiti Sains MalaysiaPenangMalaysia
  2. 2.Integrative Medicine ClusterAdvanced Medical and Dental Institute, Universiti Sains MalaysiaPenangMalaysia

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