In Vitro Cellular & Developmental Biology - Plant

, Volume 54, Issue 6, pp 621–625 | Cite as

Efficient callus-mediated regeneration and in vitro root tuberization in Trichosanthes kirilowii Maxim., a medicinal plant

  • Fenglan Zhao
  • Rong Wang
  • Jianping Xue
  • Yongbo DuanEmail author
Plant Tissue Culture


Trichosanthes kirilowii Maxim. is a climbing herb with considerable medicinal value. In this study, efficient protocols for callus-mediated regeneration and in vitro tuberization of this plant were developed. Sterilized stem and leaf tissues were cultured on Murashige and Skoog (MS) medium with plant growth regulators (PGRs), and additives that promoted callus induction and regeneration. Both stem and leaf tissues showed the best response (100%) for callus initiation on MS medium supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D). Efficient shoot organogenesis was obtained by exposing the callus tissue to 4.6-μM kinetin, 2.2-μM 6-benzylaminopurine, and 2.7-μM 1-naphthylacetic acid (NAA) along with 12.6-μM copper sulfate, which yielded a shoot regeneration rate of 85.5% and 28 shoots derived from each callus. In vitro shoots were best rooted on half-strength (1/2) MS medium with 2.7-μM NAA. Tuberous roots were efficiently induced on rooting medium with 5% (w/v) sucrose under short illumination conditions (8 h photoperiod). Rooted plantlets were successfully acclimatized in pots with a > 90% survival rate. This protocol provides an effective method for callus-mediated regeneration and in vitro root tuberization.


Trichosanthes kirilowii Callus Shoot organogenesis Copper sulfate In vitro root tuberization 


Funding information

This work was supported by the National Natural Science Foundation of China (31501368, 81573518), the Anhui Provincial Natural Science Foundation, China (1608085MC52), and the Project of Natural Science Research of Universities in Anhui Province, China (KJ2016B016).


  1. Ahmad N, Alatar AA, Faisal M, Khan MI, Fatima N, Anis M, Hegazy AK (2015) Effect of copper and zinc on the in vitro regeneration of Rauvolfia serpentina. Biol Plant 59:11–17CrossRefGoogle Scholar
  2. Cho MJ, Banh J, Yu M, Kwan J, Jones TJ (2015) Improvement of Agrobacterium-mediated transformation frequency in multiple modern elite commercial maize (Zea mays L.) inbreds by media modifications. Plant Cell Tissue Organ Cult 121:519–529CrossRefGoogle Scholar
  3. Cui L, Song J, Wu L, Huang L, Wang Y, Huang Y, Yu H, Huang Y, You CC, Ye J (2015) Smac is another pathway in the anti-tumour activity of trichosanthin and reverses trichosanthin resistance in CaSki cervical cancer cells. Biomed Pharmacother 69:119–124CrossRefGoogle Scholar
  4. Dat NT, Jin X, Hong YS, Lee JJ (2010) An isoaurone and other constituents from Trichosanthes kirilowii seeds inhibit hypoxia-inducible factor-1 and nuclear factor-kappaB. J Nat Prod 73:1167–1169CrossRefGoogle Scholar
  5. Duan YB, Zhao FL, Li H, Zhou YY, Zhu XY, Li FL, Chen WL, Xue JP (2016) Evaluation of aqueous chlorine dioxide for disinfecting plant explants. In Vitro Cell Dev Biol-Plant 52:38–44CrossRefGoogle Scholar
  6. Fan HY, Tao T, Dong SW, Li DW, Yu JL, Han CG (2013) Trichosanthes kirilowii: a new host of cucurbit mild mosaic virus in China. Plant Dis 97:1388–1388CrossRefGoogle Scholar
  7. Lee D, Seong S, Kim S, Han JB (2013) A case of stage IV non-small cell lung cancer treated with Korean medicine therapy alone. Case Rep Oncol 6:574–578CrossRefGoogle Scholar
  8. Lin GM, Li XQ, Wei HF, Li CS, Zhang JZ, Mou HF (2010) Tissue culture and rapid propagation of Trichosanthes kirilowii Maxim. J Guangdong Agri Sci 2:31–32Google Scholar
  9. Muhammad A, Rashid H, Hussain I, Naqvi SMS (2007) Proliferation-rate effects of BAP and kinetin on banana (Musa spp. AAA group) ‘Basrai’. Hortscience 5:1253–1255Google Scholar
  10. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  11. Shu S, Xie G, Guo X, Wang M (2009) Purification and characterization of a novel ribosome-inactivating protein from seeds of Trichosanthes kirilowii Maxim. Protein Expr Purif 67:120–125CrossRefGoogle Scholar
  12. Wang W, Wang L, Jiang J (2009) Fatty acid profile of Trichosanthes kirilowii Maxim seed oil. Chem Pap 63:489–492CrossRefGoogle Scholar
  13. Wu E, Lenderts B, Glassman K, Berezowskakaniewska M, Christensen H, Asmus T, Zhen SF, Chu U, Cho MJ, Zhao ZY (2014a) Optimized agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants. In Vitro Cell Dev Biol Plant 50:9–18CrossRefGoogle Scholar
  14. Wu XH, Geng MT, Fan J, Yao Y, Min Y, Li RM, Hu XW, Fu SP, Guo JC (2014b) Effects of sucrose on tuberous root formation and saccharide accumulation in Manihot esculenta Crantz in vitro. Adv Mater Res 1010-1012:225–228CrossRefGoogle Scholar
  15. Xu LY, Zhang XP (2008) Effect of different treatments on seeds germination of Trichosanthes kirilowii. J Wuhu Vocat Inst Technol 10:75–77Google Scholar
  16. Yang N, Li Z, Jiao Z, Gu P, Zhou Y, Lu L, Chou KY (2014) A trichosanthin-derived peptide suppresses type I immune responses by TLR2-dependent activation of CD8 + CD28- Tregs. Clin Immunol 153:277–287CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  • Fenglan Zhao
    • 1
  • Rong Wang
    • 1
  • Jianping Xue
    • 1
  • Yongbo Duan
    • 1
    Email author
  1. 1.Key Laboratory of Resource Plant Biology of Anhui Province, College of Life SciencesHuaibei Normal UniversityHuaibeiChina

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