Medium-Term Preservation of Trichopilia suavis Lindl. and Paxton in vitro Using Slow-Growth Technology

Muthab Hussien, Olga Molkanova and Ekaterina Raeva-Bogoslovskaya

Background and Objective: Medium-term in vitro storage using plant growth inhibitors under controlled conditions is a cost-effective method for preserving endangered plant species without frequent subculturing. This study aimed to optimize storage conditions for protocorm-like bodies (PLBs) of Trichopilia suavis, a valuable epiphytic orchid. Materials and Methods: The PLBs were cultured on half-strength Murashige and Skoog (1/2 MS) medium supplemented with varying concentrations of chlorocholine chloride (CCC) and paclobutrazol (PBZ). Cultures were incubated under two environmental conditions: A climate chamber (15±2°C, 1.7 Klux) and a culture room (24±2°C, 3.4 Klux). Data were analyzed using Analysis of Variance (ANOVA) and Duncan’s Multiple Range Test (p<0.05). Results: The PLBs cultured on 1/2 MS medium with 0.2 mg/L CCC in the climate chamber achieved 87% survival after 18 months without subculturing, with 80% regenerating into plantlets. Maximum PLB proliferation occurred with 0.4 mg/L CCC in the culture room and 1.0 mg/L PBZ in the climate chamber. The best plantlet regrowth was observed on medium with 0.5 mg/L kinetin (KIN) and coconut water. Cultivation conditions also influenced root anatomy. Roots grown in the climate chamber exhibited large diameters, a thin velamen and a thick cortex with large parenchyma cells, while roots from the culture room had thicker velamen, a more developed stele and increased xylem diameter, showing starch storage during regrowth. Conclusion: This study provides an effective protocol for medium-term preservation of T. suavis using growth inhibitors and climate chamber incubation, ensuring reliable storage of this species.

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