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Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (2): 221-231.doi: 10.7525/j.issn.1673-5102.2021.02.010

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Establishing an Efficient Regeneration Protocol in vitro for Rapid Micro-propagation of Two New Varieties of Ilex verticillata L.

Rui LIU1,2, Jia-Yi ZHANG1,2, Chao-Qi GE3, Lu-Lu XIE1,2, Ying CHEN1,2(), Can ZHANG1,2, Xi CHEN1,2   

  1. 1.Collaborative Innovation Center of the Southern Modern Forestry,Nanjing 210037
    2.College of Biology and the Environment,Nanjing Forestry University,Nanjing 210037
    3.College of 2011,Nanjing Tech University,Nanjing 211816
  • Received:2020-04-19 Online:2021-03-20 Published:2021-01-05
  • Contact: Ying CHEN E-mail:chynjfu@163.com
  • About author:LIU Rui(1997—),female,master,major in plant biotechnology and physiology.
  • Supported by:
    Science and Technology Plan Project of Jiangsu Province(BE2017375);948 Project of State Bureau of Forestry of China(2014-4-61);Program for Priority Academic Program Development of Jiangsu Province of China(PAPD);Innovation Training Program for college students of Nanjing Forestry University(2019NFUSPITP0448)


An efficient and rapid micro-propagation system using tissue culture for two new varieties “Red Sprite” and “Winter Gold” of Ilex verticillata L. was established with the young shoots as explants, and an optimum method produced large numbers of genetically uniform seedlings and maintained its excellent character for I.verticillata L was offered. The optimal medium for axillary bud induction was MS+1.0 mg·L-1 6-BA+0.2 mg·L-1 NAA, and the induction rate of both varieties were 100% respectively. The preferred medium for cluster bud proliferation was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.5 mg·L-1 CPPU. The highest proliferation coefficient was 5.7 for “Red Sprite”, and 4.5 for “Winter Gold” for 25 d culturing respectively. The optimal seedling medium for “Red Sprite” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+1.0 mg·L-1 TDZ+1.0 mg·L-1 GA, and that of “Winter Gold” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.05 mg·L-1 TDZ+1.0 mg·L-1 GA, the seedling height was up to 6.80 and 4.38 cm respectively. The optimal rooting induction medium was 1/2MS+0.4 mg·L-1 NAA+0.4 mg·L-1 IBA+1.0 mg?L-1 activated carbon, the rooting rate of “Red Sprite” and “Winter Gold” were 95.0%, or 97.0% respectively. The best transplanting substrates were peat∶perlite=3∶2, with a survival rate of 95.7% for “Red Sprite” and 96.8% for “Winter Gold” respectively.

Key words: Ilex verticillata L., varieties of “Red Sprite”, varieties of “Winter Gold”, micropropagation, an efficient regeneration protocol

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