Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 134, Issue 2, pp 183–192 | Cite as

In vitro induction of allohexaploid and resulting phenotypic variation in Populus

  • Wanting Liu
  • Yunfei Zheng
  • Shaoyu Song
  • Beibei Huo
  • Daili Li
  • Jun Wang
Original Article


Although triploid Populus varieties have been used widely in timber and pulpwood production, the performance of economic traits in Populus with higher ploidy levels remains unknown due to a lack of germplasms with higher ploidy. In this study, we first successfully induced hexaploids in Populus by treating triploid leaf explants with colchicine in vitro. In total, 32 hexaploids were produced. The frequency of hexaploids was significantly affected by the interaction between colchicine concentration and exposure time. The highest hexaploid induction efficiency was 16.89% (± 2.26), which was achieved by treating explants with 0.04% colchicine for 7 days. Compared to triploids, hexaploids had thinner epidermal hair, larger stomata and protoplasts, and fewer chloroplasts, indicating that significant phenotypic changes accompanied an increase in ploidy level. These hexaploids are valuable for investigating the performance of economic traits in Populus with higher ploidy levels and have the potential to be used as parents to produce new tetraploid and pentaploid germplasms in Populus breeding programs.


Allohexaploid Chromosome doubling Colchicine Phenotypic variation Populus Shoot organogenesis 



Murashige and Skoog (1962) basal medium


Indolebutyric acid




Thidiazuron (N-phenyl-N’-1,2,3-thiadiazol-5-ylurea)


4-Morpholinepropane sulfonate




Cell protoplast washing medium


4-Morpholine ethane sulfonic acid


Bovine serum albumin


Analysis of variance


Least significant difference


Standard error


Scanning electron microscopy


Propidium iodide



This work was supported by the National Natural Science Foundation of China (Grant No. 31470662) and the program from the Beijing Municipal Education Commission (Grant No. CEFF-PXM2018_014207_000024). The English in this document has been checked by at least two professional editors, both native speakers of English. For a certificate, please see:

Author contributions

JW conceived and designed research. WL, SS and BH conducted experiments. YZ and DL analyzed data and prepared figures. WL and JW wrote the manuscript. All authors read and approved the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.National Engineering Laboratory in Tree BreedingBeijing Forestry UniversityBeijingPeople’s Republic of China
  2. 2.Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, MOEBeijing Forestry UniversityBeijingPeople’s Republic of China
  3. 3.College of Biological Sciences and TechnologyBeijing Forestry UniversityBeijingPeople’s Republic of China
  4. 4.Beijing Huang Fa NurseryBeijingPeople’s Republic of China

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