We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Improved cryopreservation protocol for tea (Camellia sinensis (L.) O. Kuntze) using preconditioning of shoot tip donor plants and V cryo-plate technique.
- Authors
Sharma, Shailika; Mukherjee, Papiya
- Abstract
Indian tea germplasm is known for its diversity, which not only assists the development of superior planting material for tea breeding but also functions as a genetic resource for other tea-producing countries. In this study, the aim was to develop an improved cryo-protocol for long-term conservation of tea germplasm. Shoot tips (approximately 2 mm) of Camellia sinensis were cryopreserved using vitrification and V cryo-plate techniques. The recovery rate of cryopreserved shoot tips obtained using vitrification was 33.2% at 90 min of Plant Vitrification Solution 2 (PVS2) exposure. To improve recovery rates and tolerance to cryo-injury, shoot tip donor plants were preconditioned with cold hardening, high sucrose, abscisic acid (ABA), and proline prior to cryopreservation. The vitrification technique resulted in an increase of regrowth of shoot tips to 60.5% after preconditioning of shoot tip donor plants for 30 d at 25 °C with 10 mg L−1 ABA and 3 mg L−1 proline. Regrowth further increased to 75.6% with the V cryo-plate technique using shoot tip donor plants preconditioned with the above concentrations of ABA and proline. After cryopreservation, shoot tips were recovered on regrowth medium that developed into shoot cultures in 5 wk. Random amplified polymorphic DNA (RAPD) analysis was performed to test genetic fidelity after cryopreservation and results showed 100% monomorphism among all the regenerants and 10-mo-old in vitro mother cultures. Results showed that the preconditioning strategies along with V cryo-plate technique influenced and improved the cryo-recovery rate of shoot tips of tea and provided a robust protocol for tea cryopreservation.
- Subjects
TEA; RAPD technique; GERMPLASM; ABSCISIC acid; RIFLE-ranges
- Publication
In Vitro Cellular & Developmental Biology Plant, 2023, Vol 59, Issue 3, p285
- ISSN
1054-5476
- Publication type
Article
- DOI
10.1007/s11627-023-10346-4