Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 135, Issue 3, pp 445–453 | Cite as

Callus induction and plant regeneration of Spirodela polyrhiza

  • Mingxing HuangEmail author
  • Xiaoyu Ma
  • Yanshan Zhong
  • Qinxia Hu
  • Minghui Fu
  • Yali Han
Original Article


This study reports efficient protocols of tissue culture of Spirodela polyrhiza, the only species whose whole genome (including nuclear, mitochondrial and chloroplast genome) has been sequenced in Lemnaceae. The callus induction, callus maintenance and plant regeneration have been established. Sixteen kinds of combinations of phytohormones from the orthogonal combinations of four auxins and four cytokinins, and 64 kinds of orthogonal combinations of concentration for each combination of phytohormones were tested for callus induction. Callus were induced with a high efficiency from the root apical meristem on the MS medium with the phytohormone combination of 2,4-D and thidiazuron by a method called “Rhizoids-hovering”. Unlike other reported species in Lemnaceae whose callus has been induced from frond, frond of S. polyrhiza cannot be induced into callus under all tested conditions in this study. The stable callus line which is cultured on the medium of callus maintenance with reduced phytohormone under dark condition has been maintained healthy proliferation for more than three years. The callus of S. polyrhiza can be easily regenerated under a broad range of concentration of cytokinins.


Spirodela polyrhiza Tissue culture Root apical meristem Rhizoids-hovering method 



2,4-Dichlorophenoxyacetic acid


α-Naphthylacetic acid




p-Chlorophenoxy acetic acid






N6-(2-Isopentenyl) adenine







This work was supported by the Science and Technology project of Guangdong Province (No. 2016A010105020) and the Doctoral Initiation Foundation of Guangdong University of Technology (No. 253141005).

Author contributions

MXH performed most of the experiments, analyzed data, and drafted the manuscript. XYM and YSZ helped perform some experiments of tissue culture. QXH, MHF and YLH helped analyze data, draft, and revise the manuscript. All authors reviewed the results and approved the final version of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

11240_2018_1477_MOESM1_ESM.docx (257 kb)
Supplementary material 1 (DOCX 256 KB)
11240_2018_1477_MOESM2_ESM.docx (490 kb)
Supplementary material 2 (DOCX 490 KB)
11240_2018_1477_MOESM3_ESM.docx (129 kb)
Supplementary material 3 (DOCX 128 KB)


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Mingxing Huang
    • 1
    Email author
  • Xiaoyu Ma
    • 1
  • Yanshan Zhong
    • 1
  • Qinxia Hu
    • 1
  • Minghui Fu
    • 1
  • Yali Han
    • 1
  1. 1.Department of Biological Engineering, Faculty of Chemical Engineering and Light IndustryGuangdong University of TechnologyGuangzhouChina

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