Abstract
An in vitro regeneration and transient expression systems were developed for the halophyte sea aster (Aster tripolium L.), an important genetic resource for salt tolerance. Adventitious shoots were formed from both leaf explants and suspension-cultured cells in a Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) basal salts containing 500 mg l−1 casamino acids, and supplemented with 5.4 μM a-naphthaleneacetic acid (NAA) and 4.7 μM kinetin to the culture medium. Hyperhydricity of shoots was avoided by increasing the ventilation of the culture vessel. Root formation from shoots was promoted in the presence of 26.9 μM NAA. A high yield of protoplasts was isolated using 1% cellulase and 0.25% pectinase from both leaf mesophyll and suspension-cultured cells, and these were used for transient expression. The highest level of transient expression of the green fluorescent protein was obtained with 1 × 105 protoplasts ml−1, 25 μg batch−1 of plasmid vector, and 30% polyethylene glycol 4,000.
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- 2iP:
-
N6-2-Isopentenyl adenine
- BA:
-
6-Benzyladenine
- GFP:
-
Green fluorescent protein
- HSD:
-
Honestly significant difference (Tukey–Kramer’s test)
- IAA:
-
Indole-3-acetic acid
- IBA:
-
Indole-3-butyric acid
- Kn:
-
Kinetin
- MS:
-
Murashige and Skoog (1962)
- NAA:
-
a-Naphthaleneacetic acid
- PEG:
-
Polyethylene glycol
- PGR:
-
Plant growth regulator
- Zn:
-
Zeatin
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Acknowledgments
We thank Dr. Yasuo Niwa (Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka, Japan) for providing the vector CaMV35S-sGFP (S65T)-NOS (pUC19) plasmid.
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Uno, Y., Nakao, S., Yamai, Y. et al. Callus formation, plant regeneration, and transient expression in the halophyte sea aster (Aster tripolium L.). Plant Cell Tiss Organ Cult 98, 303–309 (2009). https://doi.org/10.1007/s11240-009-9564-4
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DOI: https://doi.org/10.1007/s11240-009-9564-4