Abstract
Glyptostrobus pensilis, a critically endangered plant species worldwide, exhibits low germination rate and consequently poor regeneration ability under natural conditions. In this study, sequence-related amplified polymorphism analysis shows that sexual propagation of this plant species using seeds culture could improve its genetic diversity. Germination trials of seeds at different developmental stages were performed. Seeds sampled at the immature stage in October showed the highest germination rate (19.2%) in vitro, and produced seedlings when cultured on half-strength DCR basal medium. Subsequently, the hypocotyls of seedlings were excised and incubated on callus and shoot induction medium (CSIM), comprising Murashige and Skoog (MS) basal medium supplemented with 2 mg L−1 6-benzyladenine (6-BA) and 0.5 mg L−1 α-naphthaleneacetic acid (NAA). The hypocotyl-derived callus was then used as starting material for shoot regeneration. On average, 2.4 adventitious shoots could be regenerated from a single hypocotyl-derived callus. Furthermore, more than 15 adventitious shoots were obtained from a regenerated adventitious shoot attached to a part of hypocotyl-derived callus, when sub-cultured on CSIM. After rooting and acclimation for 60 days, the plantlets were reintroduced into the conservation area, forming forests of G. pensilis. Thus, we established a circular and highly efficient adventitious shoot regeneration system of G. pensilis using hypocotyl-derived callus as the starting material.
Key Message
This study successfully obtained seedlings of G. pensilis sexual progenies via embryo culture and established a circular and highly efficient adventitious shoot regeneration system using hypocotyl-derived callus as the starting material.
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Data availability
The data sets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- 6-BA:
-
6-Benzyladenine
- CSIM:
-
Callus and shoot induction medium
- CTAB:
-
Cetyltrimethyl ammonium bromide
- dNTPs:
-
Deoxynucleotide triphosphates
- GM:
-
Germination medium
- IBA:
-
Indole-3-butyric acid
- NAA:
-
α-Naphthaleneacetic acid
- RM:
-
Rooting medium
- SRAP:
-
Sequence-related amplified polymorphism
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Acknowledgements
We thank Dr. Yingang Li of Zhejiang Academy of Forestry and Mr. Weizhong Shao of Jiande Forestry Centre for assistance in reintroduction.
Funding
This study was supported by the Zhejiang Province Key Research and Development Program (2019C02026) and the National Natural Science Foundation of China (Grant No. 31870320).
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Conceptualization, L.Y., L.X. and L.C.; Methodology, L.Y., L.X. and L.C.; Resources, L.Y., T.W., D.X., B.L. and R.C.; Investigation, L.Y., S.M., K.L. and A.Z.; Formal analysis, L.Y., S.M and L.C; Writing—Original Draft and Review & Editing, L. Y and L. C.
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Communicated by Paloma Moncaleán.
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Yuan, L., Ma, S., Liu, K. et al. High frequency adventitious shoot regeneration from hypocotyl-derived callus of Glyptostrobus pensilis, a critically endangered plant. Plant Cell Tiss Organ Cult 152, 139–149 (2023). https://doi.org/10.1007/s11240-022-02396-0
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DOI: https://doi.org/10.1007/s11240-022-02396-0