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Synthetic seeds in black huckleberry: clonal fidelity and oxidative balance assessment
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DOI：10.1007/s10725-026-01497-6.png)
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Abstract
En 中文
Ensuring genetic fidelity and proper antioxidant regulation is essential for synthetic seed technology and conservation of economically important plant species. In this study, we evaluated the effects of plant growth regulators (PGRs) and storage temperature on the germination and regeneration of encapsulated somatic embryos of Vaccinium membranaceum, while verifying genetic fidelity and physiological competence. The highest somatic embryogenesis was obtained from leaf explants cultured on berry basal medium (BM) containing 4.6 µM thidiazuron (TDZ) and 1.13 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos were encapsulated in 3% sodium alginate prepared with ½-strength BM salts and polymerized in 100 mM CaCl₂ to produce synthetic seeds. Their viability was evaluated over 12 months at − 20 °C, 4 °C and 18 °C at two-month intervals, with 4 °C identified as the most effective storage temperature for preservation. The highest germination was achieved on BM media supplemented with 4.56 µM Zeatin. Clonal fidelity of eight randomly selected synthetic seeds derived plants and donor plants were assessed using sixteen markers, all of which produced clear, reproducible monomorphic bands, confirming 100% genetic similarity with the donor plants. Antioxidant gene expression profiling showed significant upregulation of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), peroxidase (PRX) and superoxide dismutase (SOD) under in vitro conditions, while Catalase2 (CAT2) was transiently downregulated but fully recovered after acclimatization. Optimized PGRs and temperature enable efficient synthetic seed production, maintaining genetic fidelity and physiological competence, and providing a scalable platform for conservation and propagation of V. membranaceum and other woody species.
Keywords:
Vaccinium membranaceum
PGRs
Encapsulated somatic embryos
Markers
Antioxidant genes
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