Abstract
We report the successful cryopreservation of three economically important Rubus viruses: raspberry bushy dwarf virus (RBDV), black raspberry necrosis virus (BRNV), and Rubus yellow net virus (RYNV), using shoot tip cryopreservation in four raspberry cultivars. Virus-infected shoot tips (approximately 1.0 mm in length) containing 3-4 leaf primordia (LPs) were cryopreserved using the droplet-vitrification technique. In the cultivars 'Zlatá Královna (ZK)' and 'Tulameen (TUM)', over 90% of shoot tips survived, and more than 90% regenerated into whole shoots. All three viruses were successfully preserved in the cryopreserved tissues, with recovery rates varying depending on virus type and cultivar: RBDV was recovered at rates of 86% in 'ZK' and 87% in 'TUM'; BRNV at 66% in 'ZK' and 45% in 'TUM'; and RYNV at 96%, 94%, and 86% in 'Fairview', 'Stiora', and 'ZK', respectively. To investigate viral localization in shoot tips, in situ hybridization was used. RBDV and RYNV infected a broad range of meristematic tissues, including the apical dome and LPs, whereas BRNV showed a more limited distribution. Virus distribution varied not only among virus species but also across raspberry cultivars, suggesting genotype-specific patterns of virus localization. Post-cryopreservation viral activity was verified using micrografting and aphid transmission assays. RBDV, BRNV, and RYNV were all successfully transmitted to healthy plants via micrografting, indicating the preservation of viral infectivity. Furthermore, BRNV was effectively transmitted by large raspberry aphids from cryopreserved materials, confirming vector-mediated transmission capacity post-thaw. Overall, this study demonstrates that shoot tip cryopreservation via droplet-vitrification is a reliable and effective strategy for preservation of biologically active Rubus viruses. This approach offers a valuable biotechnological tool for virus maintenance in support of diagnostic, breeding, and virology research.
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Claims 15
Claim · model-written summary
More than 90% of cryopreserved Zlatá Královna shoot tips survived.
Verbatim
e0“In the cultivars 'Zlatá Královna (ZK)' and 'Tulameen (TUM)', over 90% of shoot tips survived”
t0“raspberry shoot tips”
Claim · model-written summary
More than 90% of cryopreserved Tulameen shoot tips survived.
Verbatim
e0“In the cultivars 'Zlatá Královna (ZK)' and 'Tulameen (TUM)', over 90% of shoot tips survived”
t0“raspberry shoot tips”
Claim · model-written summary
More than 90% of shoot tips in Zlatá Královna regenerated into whole shoots.
Verbatim
e0“In the cultivars 'Zlatá Královna (ZK)' and 'Tulameen (TUM)', over 90% of shoot tips survived, and more than 90% regenerated into whole shoots”
t0“raspberry shoot tips”
Claim · model-written summary
More than 90% of shoot tips in Tulameen regenerated into whole shoots.
Verbatim
e0“In the cultivars 'Zlatá Královna (ZK)' and 'Tulameen (TUM)', over 90% of shoot tips survived, and more than 90% regenerated into whole shoots”
t0“raspberry shoot tips”
Claim · model-written summary
RBDV recovery was 86% in ZK.
Verbatim
e0“RBDV was recovered at rates of 86% in 'ZK'”
t0“raspberry shoot tips”
Claim · model-written summary
RBDV recovery was 87% in TUM.
Verbatim
e0“RBDV was recovered at rates of 86% in 'ZK' and 87% in 'TUM'”
t0“raspberry shoot tips”
Claim · model-written summary
BRNV recovery was 66% in ZK.
Verbatim
e0“BRNV at 66% in 'ZK'”
t0“raspberry shoot tips”
Claim · model-written summary
BRNV recovery was 45% in TUM.
Verbatim
e0“BRNV at 66% in 'ZK' and 45% in 'TUM'”
t0“raspberry shoot tips”
Claim · model-written summary
RYNV recovery was 96% in Fairview.
Verbatim
e0“RYNV at 96%, 94%, and 86% in 'Fairview', 'Stiora', and 'ZK', respectively”
t0“raspberry shoot tips”
Claim · model-written summary
RYNV recovery was 94% in Stiora.
Verbatim
e0“RYNV at 96%, 94%, and 86% in 'Fairview', 'Stiora', and 'ZK', respectively”
t0“raspberry shoot tips”
Claim · model-written summary
RYNV recovery was 86% in ZK.
Verbatim
e0“RYNV at 96%, 94%, and 86% in 'Fairview', 'Stiora', and 'ZK', respectively”
t0“raspberry shoot tips”
Claim · model-written summary
RBDV and RYNV infected the apical dome and leaf primordia, while BRNV had a more limited distribution.
Verbatim
e0“RBDV and RYNV infected a broad range of meristematic tissues, including the apical dome and LPs, whereas BRNV showed a more limited distribution”
t0“raspberry shoot tips”
Claim · model-written summary
Virus localization differed across raspberry cultivars.
Verbatim
e0“Virus distribution varied not only among virus species but also across raspberry cultivars”
Claim · model-written summary
RBDV, BRNV and RYNV retained infectivity and were transmitted to healthy plants by micrografting.
- Taxon
- raspberryt0
- Interactor
- RBDVRaspberry bushy dwarf viruse0pathogen
- Second interactor
- BRNVe0pathogen
- Outcome
- successfully transmitted to healthy plants via micrograftinge0
- Qualifier
- RYNVe0
Not reportedValue
Verbatim
e0“RBDV, BRNV, and RYNV were all successfully transmitted to healthy plants via micrografting, indicating the preservation of viral infectivity”
t0“raspberry shoot tips”
Claim · model-written summary
BRNV was transmitted by large raspberry aphids from cryopreserved material after thawing.
- Taxon
- raspberrye0
- Interactor
- BRNVe0pathogen
- Second interactor
- large raspberry aphidsAmphorophora idaeie0pest
- Outcome
- vector-mediated transmission capacity post-thawe0
Not reportedValue
Verbatim
e0“BRNV was effectively transmitted by large raspberry aphids from cryopreserved materials, confirming vector-mediated transmission capacity post-thaw”