Micropropagation of Pinus densiflora and the evaluation of nematode resistance of regenerated microshoots in vitro

  • Lihua Zhu
  • Xiaofang Chu
  • Tingyu Sun
  • Jianren Ye
  • Xiaoqin Wu
Original Paper


To accelerate the breeding and selection of Pinus densiflora Siebold and Zucc. resistance to pine wilt disease, a micropropagation system was established and nematode resistance evaluated in vitro. Cotyledon-hypocotyl explants from 28-day-old seedlings were first cultured on Gresshoff and Doy medium supplemented with 4.0 mg L−1 6-benzyladenine and 0.2 mg L−1 α-naphthaleneacetic acid (NAA) to stimulate the formation of buds. Induced buds were subsequently subcultured on Gupta and Durzan medium supplemented with 0.1% (w/v) activated charcoal for elongation. Stem sections derived from shoots were used as explants for the further multiplication. Roots were formed from shoots transferred to woody plant medium containing 0.2 mg L−1 NAA for 4 weeks. The nematode resistance test showed that symptoms in micropropagated shoots after infection with pine wood nematode (PWN) were similar to those in plants infected in the field. The wilting rate varied from 20 to 100% among different clones 18 days after inoculation. The most susceptible clone was Clone 6-4 with a 100% wilting rate, while Clone 8-4 showed a relatively high resistance with a 20% wilting rate. The number of nematodes recovered from Clone 8-4 shoots was significantly lower (P = 0.05) than from Clones 5-10 and 16-4. This work contributes to the breeding of PWN resistance in P. densiflora.


Pinus densiflora Micropropagation Shoot proliferation Bursaphelenchus xylophilus Nematode resistance 



We thank the reviewers for their help and invaluable advice in preparing this manuscript.


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Copyright information

© Northeast Forestry University and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Lihua Zhu
    • 1
    • 2
  • Xiaofang Chu
    • 1
    • 2
  • Tingyu Sun
    • 1
    • 2
  • Jianren Ye
    • 1
    • 2
  • Xiaoqin Wu
    • 1
    • 2
  1. 1.Institute of Forest Protection, College of ForestNanjing Forestry UniversityNanjingPeople’s Republic of China
  2. 2.Co-Innovation Center for Sustainable Forestry in Southern ChinaNanjingPeople’s Republic of China

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