Year 2025, Volume 10, Issue 3
| Year : | 2025 |
| Volume : | 10 |
| Issue : | 3 |
| Authors : | Natalia ONICA, Ioana-Cătălina NICOLAE, Ion ROȘCA, Maria TABĂRA, Nina CIORCHINĂ, Aliona GLIJIN, Oana VENAT |
| Title : | GENOTYPE-DEPENDENT RESPONSES TO STERILIZATION AND RESTERILIZATION IN IN VITRO CULTURES OF HYDRANGEA MACROPHYLLA |
| Abstract : | Establishing axenic cultures of Hydrangea macrophylla remains a significant challenge due to persistent endogenous contamination. This study evaluated complex disinfection and resterilization protocols for three cultivars:”Love”, ”Tivoli®”, and ”Masja”. Explants (apical/lateral buds) were subjected to multi-step sterilization involving ethanol, sodium dichloroisocyanurate (NaDCC, 0.2 - 2%), permanganate, and fungicidal (Dithane®) or bleach-based disinfectant (Domestos®) - commercial bleach. Inoculations were done on MS medium with BAP or BAP + kinetin, and cultures were maintained at 23 - 25°C, 2000 - 2500 lux, 16 h photoperiod. Initial contamination exceeded 80% for all genotypes. Weekly resterilizations (up to three cycles) using NaDCC (2%) + 3 drops Domestos reduced bacterial and fungal contamination by ~45% in ‘Masja’ and ~35% in ”Love”, but only marginally (~10%) in ”Tivoli®”. Fungal suppression was more effective when Dithane® (0.2%) was included for 7 minutes prior to NaDCC treatment. ”Tivoli®” explants exhibited persistent internal bacterial symptoms despite surface sterilization, confirming its status as the most recalcitrant genotype. Survival and meristem establishment rates were highest in ”Masja” (34%) and lowest in ”Tivoli®” (8%). Callus induction occurred in up to 40% of clean explants. Multiple rinses (3×2 min in sterile water) and intermediate permanganate exposure (max. 15 minutes) proved critical in improving aseptic conditions. The study demonstrates that decontamination efficiency is highly genotype-dependent and supports a tailored, iterative approach to resterilization. Results provide practical guidance for establishing stable in vitro cultures of Hydrangea macrophylla cultivars for conservation and micropropagation. |
| For citation : | Onica, N., Nicolae, I.-C., Roșca, I., Tabăra, M., Ciorchină, N., Glijin, A., Venat, O. (2025). Genotype-dependent responses to sterilization and resterilization in in vitro cultures of Hydrangea macrophylla. AGROFOR International Journal, Volume 10. Issue No. 3. pp. 48-56. DOI:10.7251/AGREN2503048O |
| Keywords : | Contamination, Meristem culture, Disinfection protocol. |
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ISSN 2490-3434 (Printed)
ISSN 2490-3442 (Online)
ISSN 2490-3442 (Online)
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