Effect of Encapsulation Parameters and Storage Conditions on Synthetic Seeds of Vanda bensonii Batemen

Abstract

The present study investigates the medium-term storage of regenerated plants of Vanda bensonii Batemen using synthetic seed technology, wherein protocorm-like bodies (PLBs) were encapsulated in sodium alginate and complexed with calcium chloride to produce bead-like propagules capable of regenerating into complete plantlets. Bead morphology was influenced by the concentrations of sodium alginate and calcium chloride, with 3% (w/v) sodium alginate and 100 mM CaCl₂ producing mostly globular, tail-free beads that ensured good encapsulation integrity and handling efficiency. Increasing CaCl₂ concentration to 200 mM led to uneven cross-linking and distorted bead shapes, indicating that 100 mM CaCl₂ offers near-optimal gelation conditions. The regeneration potential of encapsulated PLBs was assessed after storage at different temperatures and durations. At 4°C, the PLBs exhibited high viability, with regeneration rates of 95.16% at 15 days, 92.73% at 30 days, and 85.71% at 45 days, followed by a gradual decline over longer storage periods. Although a slight recovery was observed at 75 days (90.16%), viability significantly decreased after six months, reaching 53.33% at 270 days. Conversely, storage at 16°C resulted in rapid deterioration, with regeneration falling from 36.36% at 15 days to complete loss beyond 45 days. These results demonstrate that low-temperature storage (4°C) is markedly superior for maintaining the viability and regeneration potential of synthetic seeds by slowing metabolic degradation.

Plant Tissue Cult. & Biotech. 36(1): 139-147, 2026 (June)