Callus induction and plant regeneration of Spirodela polyrhiza
- 605 Downloads
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.
KeywordsSpirodela polyrhiza Tissue culture Root apical meristem Rhizoids-hovering method
p-Chlorophenoxy acetic acid
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).
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.
- Abhishek A, Karjagi CG, Nath R, Bhardwaj M, Ramteke PW, Kumar P, Dass S, Kumar RS (2014) Differential effect of immature embryo’s age and genotypes on embryogenic type II callus production and whole plant regeneration in tropical maize inbred lines (Zea mays L.). Indian J Genet Plant Breed 74:317–324CrossRefGoogle Scholar
- Islam K (2002) Feasibility of duckweed as poultry feed—a review. India J Anim Sci 72:486–491Google Scholar
- Wang WQ, Kerstetter RA, Michael TP (2011) Evolution of genome size in duckweeds (Lemnaceae). J Bot 1:9Google Scholar
- Wang W, Wu Y, Messing J (2012) The mitochondrial genome of an aquatic plant, Spirodela polyrhiza. PLOS ONE 7:e4674710Google Scholar
- Wang W, Haberer G, Gundlach H, Glaesser C, Nussbaumer T, Luo MC, Lomsadze A, Borodovsky M, Kerstetter RA, Shanklin J, Byrant DW, Mockler TC, Appenroth KJ, Grimwood J, Jenkins J, Chow J, Choi C, Adam C, Cao XH, Fuchs J, Schubert I, Rokhsar D, Schmutz J, Michael TP, Mayer KFX, Messing J (2014) The Spirodela polyrhiza genome reveals insights into its neotenous reduction fast growth and aquatic lifestyle. Nat Commun 5:1–14Google Scholar