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Somatic embryogenesis and cryopreservation of Stewartia species

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Abstract

Stewartia is a genus of flowering shrubs and trees in the family Theaceae. Stewartia species are native to eastern Asia, with the exception of S. ovata and S. malacodendron, which are indigenous to southeastern North America. Despite having outstanding ornamental value and unique horticultural features, Stewartias are not readily available for landscaping in the horticultural trade due to difficulty with mass propagation. In addition to commercial propagation, there is a need to develop propagation techniques for the conservation of the North American species, which are rare (S. ovata) or endangered (S. malacodendron). This research examined somatic embryogenesis as a means to mass propagate Asian and North American Stewartia species and to provide material for cryopreservation of valuable Stewartia germplasm. By testing a variety of plant growth regulators (PGRs), somatic embryogenesis was induced from immature zygotic embryo explants of seven Stewartia species—five Asian and two North American. Additionally, embryogenic cultures were obtained from three North American cultivars, S. malacodendron ‘Delmarva’, S. ovata ‘Red Rose’, and ‘Royal Purple’. For two Stewartia species, S. ovata and S. sinensis, somatic embryogenesis was induced from immature endosperm. Picloram (0.05 or 0.1 mg/L) and 2,4-dichlorophenoxyacetic (2 or 4 mg/L) were the most effective PGRs for inducing somatic embryogenesis of North American and Asian Stewartia species, respectively. All seven Stewartia species examined produced somatic embryos that converted into plants that could be transferred to the greenhouse. Embryogenic tissue from five Stewartia species were successfully put into cryo-storage and recovered, providing a means of long-term germplasm storage. This is the first report of somatic embryogenesis in the genus Stewartia.

Key message

This is the first report of somatic embryogenesis from immature zygotic embryos in the genus Stewartia and it was accomplished with seven different species.

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Abbreviations

BAP:

6-Benzylaminopurine

IBA:

Indole-3-butyric acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

Picloram:

4-Amino-3,5,6-trichloro-2-pyridinecarboxylic acid

meta-Topolin:

6-(3-Hydroxybenzylamino)purine

NAA:

1-Naphthaleneacetic acid

DMSO:

Dimethyl sulfoxide

MS:

Murashige and Skoog medium

WPM:

Woody Plant Medium

EMM:

Embryo maturation medium

PGRs:

Plant Growth Regulators

PEMs:

Pro-embryogenic masses

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Acknowledgements

The authors are grateful to John Clark and Tim Boland from The Polly Hill Arboretum for providing plant material of their rare cultivars of S. ovata and S. malacodendron and to Bill Hawkins for plant material of S. malacodendron from his property in Sylvania, GA. This work received the financial support from the Warnell School of Forestry and Natural Resources, University of Georgia and from the Georgia Native Plant Society.

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Authors and Affiliations

  1. Department of Horticulture, University of Georgia, 1111 Plant Sciences Building, Athens, GA, 30602, USA

    Heather J. Gladfelter & H. Dayton Wilde

  2. P.O. Box 121, Lakemont, GA, 30552, USA

    Jack Johnston

  3. Warnell School of Forestry & Natural Resources, University of Georgia, 180 E. Green St., Athens, GA, 30602, USA

    Heather J. Gladfelter & Scott A. Merkle

Authors
  1. Heather J. Gladfelter
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  2. Jack Johnston
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  3. H. Dayton Wilde
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  4. Scott A. Merkle
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Contributions

HJG conceived, conducted experiments, analyzed data, and wrote the manuscript. HDW and SAM contributed to the conception, organizing and editing of the manuscript. JJ provided immature fruit of seven different Stewartia species and shared his expertise in propagating and growing Stewartia.

Corresponding author

Correspondence to Heather J. Gladfelter.

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The authors declare that they have no conflict of interest.

Additional information

Communicated by Qiao-Chun Wang.

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Electronic supplementary material

Below is the link to the electronic supplementary material.

Online Resource 1. Chi-Square analysis for S. malacodendron ‘Delmarva’ in Table 1. Supplementary file1 (DOCX 48 kb)

Online Resource 2. Chi-Square analysis for S. ovata ‘Red Rose’ in Table 1. Supplementary file2 (DOCX 48 kb)

Online Resource 3. Chi-Square analysis for S. rostrata in Table 1. Supplementary file3 (DOCX 47 kb)

Online Resource 4. Chi-Square analysis for S. ovata ‘Royal Purple’ in Table 2. Supplementary file4 (DOCX 51 kb)

Online Resource 5. Chi-Square analysis for S. sinensis in Table 2. Supplementary file5 (DOCX 46 kb)

Online Resource 6. Chi-Square analysis for S. malacodendron ‘Delmarva’ in Table 2. Supplementary file6 (DOCX 43 kb)

Online Resource 7. Chi-Square analysis for S. monadelpha in Table 2. Supplementary file7 (DOCX 53 kb)

Online Resource 8. Chi-Square analysis for S. pseudocamellia var. koreana in Table 2. Supplementary file8 (DOCX 53 kb)

Online Resource 9. Chi-Square analysis for S. rostrata in Table 3. Supplementary file9 (DOCX 53 kb)

11240_2020_1834_MOESM10_ESM.docx

Online Resource 10. Chi-square analysis for the induction of somatic embryogenesis from the zygotic embryos of S. ovata ‘Otto, NC’. Supplementary file10 (DOCX 51 kb)

11240_2020_1834_MOESM11_ESM.docx

Online Resource 11. Chi-square analysis for the induction of somatic embryogenesis from the zygotic endosperm of S. ovata ‘Otto, NC’. Supplementary file11 (DOCX 53 kb)

Online Resource 12. Chi-Square analysis for S. sinensis in Fig. 4. Supplementary file12 (DOCX 51 kb)

Supplementary file13 (XLSX 19 kb)

Supplementary file14 (XLSX 26 kb)

Supplementary file15 (XLSX 11 kb)

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Gladfelter, H.J., Johnston, J., Wilde, H.D. et al. Somatic embryogenesis and cryopreservation of Stewartia species. Plant Cell Tiss Organ Cult 144, 211–221 (2021). https://doi.org/10.1007/s11240-020-01834-1

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  • DOI: https://doi.org/10.1007/s11240-020-01834-1

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