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Callus induction, plant regeneration, and long-term maintenance of embryogenic cultures in Zoysia matrella [L.] Merr

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Abstract

An efficient protocol of callus induction, plant regeneration and long-term maintenance of embryogenic cultures for manilagrass was developed. Callus induction and embryogenic callus formation were influenced by cytokinins and nodal positions. Murashige and Skoog (MS) medium with 2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.02 mg l−1 kinetin (KT) or 6-benzyladenine (BA) gave the highest frequency for both callus induction and embryogenic callus formation compared with 0.02 mg l−1 thidiazuron (TDZ) or N6-(2-isopenteny) adenine (2iP). The frequency of callus induction of different nodes (from the first to the sixth node) varied from 22.5 to 92.1%, and the embryogenic callus formation frequencies ranged from 13.3 to 25.7%. The highest frequencies of callus induction and embryogenic callus formation (92.1 and 25.7%, respectively) were observed in the fourth node group. During subculture on callus induction and maintenance medium, somatic embryos formed on the surface of the embryogenic callus. On regeneration medium, the regeneration rates of embryogenic callus varied from 96.8 to 100% during the 4-year period of subculture. The results also indicate that preservation of manilagrass callus is stable at low-temperature (4°C) over a period of 11 months. No significant differences were found in the activities of superoxide dismutase (SOD), peroxidase (POD) and proline content of the plants regenerated from the 4-year subcultured callus on different regeneration media.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BA:

6-Benzyladenine

ZT:

Zeatin

KT:

Kinetin

TDZ:

Thidiazuron

2iP:

N6-(2-isopenteny) adenine

MS:

Murashige and Skoog

SOD:

Superoxide dismutase

POD:

Peroxidase

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Acknowledgments

This study has been carried out with financial support from National Science Foundation of China (30771522). Mr. Fanlong Kong and Ms. Xia Cao are acknowledged for their assistance with the lab work, and Dr. Shirley Burgess is acknowledged for revising the English in the manuscript.

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Authors and Affiliations

  1. Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Huajiachi Campus, 310029, Hangzhou, People’s Republic of China

    Mingliang Chai, Yufang Jia, Shu Chen, Zhongshan Gao, Hefei Wang, Lulu Liu, Peijia Wang & Daqiang Hou

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  1. Mingliang Chai
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  2. Yufang Jia
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  3. Shu Chen
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  4. Zhongshan Gao
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  5. Hefei Wang
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  6. Lulu Liu
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  7. Peijia Wang
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  8. Daqiang Hou
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Correspondence to Mingliang Chai.

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Chai, M., Jia, Y., Chen, S. et al. Callus induction, plant regeneration, and long-term maintenance of embryogenic cultures in Zoysia matrella [L.] Merr. Plant Cell Tiss Organ Cult 104, 187–192 (2011). https://doi.org/10.1007/s11240-010-9817-2

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