Abstract
Slash pine (Pinus elliottii Engelm.) has strong adaptability, early growth and high turpentine yield, and it is widely planted in southern China. Breeding of disease-resistant varieties is an effective way to prevent and control development of pine needle brown spot on slash pine in China. Plant regeneration by somatic embryogenesis has been achieved for slash pine, but large-scale production of somatic embryos remains restricted by several factors. We tested different embryogenesis conditions from time of initiation to maturation and studied genetic stability at simple sequence repeat (SSR) loci of regenerated plants in slash pine. Immature zygotic embryos of four open-pollinated mother trees were used for initiation of somatic embryogenesis. Seed sources (families) had a significant impact on initiation of embryonal suspensor mass (ESM, p < 0.05); however, the addition of abscisic acid ABA (1–2 mg/L) and phytosulfokine PSK (0.5 mg/L) could effectively improve initiation rate by up to 36%. Seed family 27 was the most favorable female parent for ESM initiation. The production (number of somatic embryos) was significantly increased by adding ABA (5 mg/L) during suspension culture before transferring to maturation media (p < 0.05). We tested the genetic stability of 14 regenerated plants at SSR loci, and 88.9% plants with normal phenotypes were found not to have genetic variation. We clarified the somatic embryogenesis conditions suitable for slash pine and established somatic embryo maturation technology.
Key message
We clarified the somatic embryogenesis conditions suitable for slash pine and established somatic embryo maturation technology by studying several related influencing factors.
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- KT:
-
Kinetin
- BA:
-
6-Benzylaminopurine
- NAA:
-
1-Naphthylacetic acid
- ABA:
-
Abscisic acid
- IBA:
-
Indole-3-butyric acid
- ESM:
-
Embryonal suspensor mass
- ECL:
-
Embryogenic cell line
- PGR:
-
Plant growth regulator
- FG:
-
Female gametophyte
- PEM:
-
Proembryogenic masses
- PSK:
-
Phytosulfokine
- SSR:
-
Simple sequence repeat
- RAPD:
-
Random amplified polymorphic DNA
- RFLP:
-
Restriction fragment length polymorphism
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Acknowledgements
This research was financially supported by the National Key Research and Development Program of China (No. 2017YFD0600104), the National Natural Science Foundation of China (No. 31971659) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD). Thanks are due to Xi’po State-owned Forest Farm for supplying pine cones. We thank International Science Editing (http://www.internationalscienceediting.com) for editing this manuscript.
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FY and LHZ developed the idea of the study, participated in its design and coordination and helped to draft the manuscript. XRX and XK contributed to the acquisition and interpretation of data. LHZ and JRY revised the manuscript. All authors read and approved the final manuscript.
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Yang, F., Xia, XR., Ke, X. et al. Somatic embryogenesis in slash pine (Pinus elliottii Engelm): improving initiation of embryogenic tissues and maturation of somatic embryos. Plant Cell Tiss Organ Cult 143, 159–171 (2020). https://doi.org/10.1007/s11240-020-01905-3
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DOI: https://doi.org/10.1007/s11240-020-01905-3