Abstract
In Dalbergia latifolia Roxb. (Indian rosewood), a premium-quality vulnerable timber tree species belonging to Family Fabaceae, direct somatic embryogenesis was achieved. Immature cotyledons from green pods were collected from trees at 90 d after flowering and somatic embryo formation was achieved on Murashige and Skoog’s medium supplemented with 0.5 mg L−1 2,4-dichlorophenoxyacetic acid and 0.1 mg L−1 kinetin. Different developmental stages, ranging from globular, heart-shaped, torpedo, and cotyledonary stage, were recorded during the complete formation of somatic embryos. Globular somatic embryos gradually converted into heart shape after 35 to 40 d, torpedo shape after 50 to 55 d, and dicotyledonary-shaped embryos after 60 d of culture. The individual isolated somatic embryos with well-developed cotyledons were transferred on medium containing 0.5 mg L−1 benzyl adenine and 0.1 mg L−1 abscisic acid to promote development into plantlets. Biochemical changes in soluble sugars, total carbohydrates, phenols, and nitrate reductase activity were monitored at 5-d intervals from 0 to 60 d. Soluble sugar content (%) increased from 5.26% to 28.05% during the culture period. Minimum sugar content was noted in the globular stage, followed by heart-shaped and torpedo stage and maximum sugar content was observed in cotyledonary somatic embryo stage. A similar trend was observed in total carbohydrate content also (12.83% to 41.23%). A significant variation in the phenol content and nitrate reductase activity of the somatic embryos was observed and the peak values were achieved at 35 d in phenol content (4.25%) and 30 d in nitrate reductase activity (0.79 μmol NO2 h−1 g−1). Therefore, it was concluded that increasing levels of endogenous soluble sugar and carbohydrate play an important role during the formation of somatic embryos. These findings can be utilized to study the underlying carbohydrate and nitrogen metabolic pathways during somatic embryogenesis.
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Acknowledgments
The authors gratefully acknowledge the Director and Group Coordinator Research of Tropical Forest Research Institute, Jabalpur for providing all necessary facilities for the study. The help and support from all members of Genetics and Tree Improvement Division is also thankfully acknowledged. The support provided in forest areas by Madhya Pradesh, Chhattisgarh, Maharashtra, and Odisha State Forest Departments in the course of project work is also gratefully acknowledged.
Funding
The project was financially supported by Indian Council of Forestry Research and Education Dehradun, India, under project ID No. 204/TFRI/2013/Gen-1(28) (2013–2017).
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Shirin, F., Bhadrawale, D., Mishra, J.P. et al. Evaluation of biochemical changes during different stages of somatic embryogenesis in a vulnerable timber tree Dalbergia latifolia (Indian rosewood). In Vitro Cell.Dev.Biol.-Plant 56, 894–902 (2020). https://doi.org/10.1007/s11627-020-10099-4
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DOI: https://doi.org/10.1007/s11627-020-10099-4