Grapevine Shoot Tip Cryopreservation and Cryotherapy: Secure Storage of Disease-Free Plants
Abstract
:1. Introduction
2. Explant Sources
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
---|---|---|---|---|---|---|---|
V. labrusca, 3 | ST (1–2 mm; type n/s) harvested from greenhouse plants | TEC | None | None | 10% DMSO + 60 g L−1 sucrose (2 h at 20 °C) → cooled (0.5 °C min−1) to −20 °C/−30 °C/−40 °C → LN | 87–100% survival | 1989 [84] |
V. vinifera, 1 | AxST (size n/s) harvested from in vitro cultures that are 7 weeks old | ED + TEC | None | Liquid MS with increased sucrose concentrations every 2 d of 0.3, 0.5 and 0.75 M every 2 d, and then every 1 day of 1, 1.25 and 1.5 M (temperature n/s) | Bead desiccation to 20% + cooled (0.5 °C min−1) from +20 °C to −80 °C → LN | 24 | 1991 [85] |
ED | Bead desiccation to 20% → LN | No shoot regrowth | |||||
V. vinifera, 1 | AxST (size n/s) harvested from in vitro cultures that are 7 to 8 weeks old | ED + TEC | None | Liquid MS with increased sucrose concentrations every 2 days of 0.3, 0.5 and 0.75 M and 1 M (temperature n/s) | Bead desiccation to 22% + cooled (0.5 °C min−1) from + 20 °C to −100 °C → LN | 30 | 1993 [86] |
ED | Bead desiccation to 22% → LN | 30% survival | |||||
V. vinifera, 3 | AxST (size n/s) harvested from in vitro cultures (age n/s) | ED + TEC | None | Liquid MS with increased sucrose concentrations every 24 h of 0.3, 0.6 and 1 M (25 °C) | Bead desiccation to 30% + beads cooled from 0.5 °C min−1 to −80 °C → LN | No shoot regrowth | 2000 [87] |
LN33 (Vitis L.), 1; V. vinifera, 1 | ST (1 mm; type n/s) harvested from in vitro cultures that are 4 weeks old | ED | None | 1/2 MS with increased sucrose concentrations every 24 h of 0.25, 0.5, 0.75 and 1 M + 2.6 g L−1 gellan gum (24 °C) | Bead desiccation to 16% → LN | 40–60% survival | 2000 [88] |
V. vinifera, 4 | AxST (2 mm) harvested from in vitro cultures that are 5 months old | ED + TEC | Cold-hardening for 4 weeks at 5 °C | B5 medium with increased sucrose concentrations every 24 h of 0.1, 0.3, 0.7 and 1 M + 5 g L−1 agar (5 °C) | Bead desiccation to 21% + cooled (0.2 °C min−1) to −40 °C → LN | 15–40 | 2001 [89] |
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
LN33 (Vitis L.), 1 | ST (1 mm; type n/s) harvested from in vitro cultures that are 4 weeks old | VI | None | 1/2 MS with increased sucrose concentrations every 24 h of 0.25, 0.5 and 0.75 M + 2.6 g L−1 Gelrite (24 °C) | 2 M glycerol + 0.75 M sucrose (60 min at 25 °C) → 1/2 PVS2 (30 min at 0 °C) → PVS2 (50 min at 0 °C) → LN | 45% survival | 2003 [33] |
ED | Preculture described above + additional 1 d on 1 M sucrose + 2.6 g L−1 Gelrite | Bead desiccation by air-drying for 8 h (beads moisture content n/s) → LN | 63% survival | ||||
V. vinifera, 1 | ST (1 mm; type n/s) harvested from in vitro cultures that are 4 weeks old | VI | None | 1/2 MS with increased sucrose concentrations every 24 h of 0.25, 0.5 and 0.75 M + 2.6 g L−1 Gelrite (24 °C) | 2 M glycerol + 0.75 M sucrose (60 min at 25 °C) → 1/2 PVS2 (30 min at 0 °C) → PVS2 (50 min at 0 °C) → LN | 50% survival | 2003 [64] |
ED | Preculture described above + additional 1 day on 1 M sucrose + 2.6 g L−1 Gelrite | Bead desiccation by air-drying for 7 h (beads moisture content n/s) → LN | 62% survival | ||||
V. berlandieri x V. riparia, 1 | AST and AxST (1–2 mm) harvested from in vitro cultures (age n/s) | EN-VI | Cold-hardening for 3 weeks at 4 °C | None | PVS2 (30 and 90 min at 0 °C) → LN | Low (n/s) | 2003 [90] |
V. vinifera, 7; V. berlandieri x riparia, 2; V. mourvedre × V. rupestris, 1; V. coignetiae, 1 | AxST (1 mm) harvested from in vitro cultures that are 4 to 5 months old | VI | None | 1/2 MS + 0.3 M sucrose + 2 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 25 °C) → 1/2 PVS2 (30 min at 0 °C) → PVS2 (50 min at 0 °C) → LN | 30–87 | 2000 [31] 2003 [32] |
V. vinifera, 4 | AST (2 mm) harvested from in vitro cultures that are 50 days old | ED + TEC | None | Culture medium with increased sucrose concentrations every 24 h of 0.5, 0.70 and 1 M (5 °C) | Bead desiccation to 26% + cooled (0.2 °C min−1) from 0 °C to −40 °C → LN | 36 (average) | 2003 [91] |
V. berlandieri × riparia, 1 | AxST (size n/s) harvested from in vitro cultures (age n/s) | VI | None | 0.2, 0.3 or 0.4 M sucrose (duration and conditions n/s) | 2 M glycerol + 0.4 M sucrose (30 min) → PVS2 (30, 60 or 90 min) (conditions n/s) → LN | No shoot regrowth | 2007 [92] |
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
V. vinifera, 1 | ST (1 mm; type n/s) harvested from shoots of greenhouse plants | ED | None | 3/4 MS with increased sucrose concentrations every 24 h of 0.25, 0.5, 0.75 and 1 M (conditions n/s) | Bead desiccation by air-drying for 12 h (beads moisture content n/s) → LN | 59 | 2011 [63] |
V. vinifera, 1 | ST (2–3 mm; type n/s) harvested from in vitro cultures (age n/s) | VI | None | MS + 0.3 M sucrose + 8 g L−1 agar for 1 day at 24 °C | 5% (w/v) DMSO + 5% (w/v) glycerol + 5% (w/v) sucrose (50 min at 0 °C) → PVS2 (40 min at 0 °C) or MPVS2 **** (40 min at 0 °C) → LN | 47 and 55 | 2011 [93] |
V. berlandieri × V. riparia, 1 | AxST (2 ± 1 mm) harvested from in vitro cultures (age n/s) | VI | Cold-hardening for 2 weeks at 4 °C | MS + 0.2, 0.3 or 0.4 M sucrose + 8 g L−1 agar for 2 days at 4 °C | 2 M glycerol + 0.4 M sucrose (30 min at 4 °C) → PVS2 (30, 60 or 90 min at 0 °C) → LN | No shoot regrowth | 2012 [94] |
V. vinifera, 1 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | ED | Nodal sections on 1/2 MS + Morel’s vitamins * + 20 g L−1 sucrose + 1 μmol ZR + 7 g L−1 agar for 2 weeks at 24 °C | Liquid 1/2 MS with increased sucrose concentrations every 12 h of 0.25, 0.5, 0.75 and 1 M (24 °C) | Bead desiccation to 22.3% → LN | 37 | 2013 [95] |
DV | MS + 1 M sucrose + 7 g L−1 agar for 24 h at 24 °C | 2 M glycerol + 0.4 M sucrose (20 min at RT) → PVS2 (50 min at 0 °C) → LN | 50 | ||||
V. vinifera, 2 | AxST (1 mm) harvested from greenhouse-grown plants | DV | None | 1/2 MS + 0.3 M sucrose + 0.16 mM GSH reduced + 0.14 mM AsA + 2.5 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (10–15 min at 0 °C) → PVS2 (10–20 min at 0 °C) → LN | 40–46 | 2013 [96] |
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
V. vinifera, 1 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | Nodal sections on 1/2 MS + Morel’s vitamins * + 20 g L−1 sucrose + 1 μmol BA or ZR + 7 g L−1 agar for 2 weeks at 24 °C | 1/2 MS + 0.1 M sucrose + 7 g L−1 agar for 24 h at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at RT) → 1/2 PVS2 (30 min at RT) → PVS2 (50 min at 0 °C) → LN | 44 | 2014 [97] |
AxST (1 mm) harvested from in vitro cultures that are 2 months old | None | 2 M glycerol + 0.4 M sucrose (20 min at RT) → 1/2 PVS2 (30 min at RT) → PVS2 (75 min at 0 °C) → LN | 41.6 | ||||
V. vinifera, 1 | AxST (size n/s) harvested from vineyard | VI | None | None | PVS2 (180 min at 25 °C) → LN | n/s | 2015 [98] |
V. vinifera, 9 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | Nodal sections on 1/2 MS + Morel’s vitamins * + 20 g L−1 sucrose + 1 μmol BA + 7 g L−1 agar for 2 weeks at 24 °C | 1/2 MS + 0.1 M sucrose + 7 g L−1 agar for 24 h at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at RT) → 1/2 PVS2 (30 min at RT) → PVS2 (50 min at 0 °C) → LN | 0–70 | 2015 [34] |
V. vinifera, 7; V. labrusca, 1; V. riparia, 1; V. berlandieri × V. rupestris, 2; V. berlandieri × V. riparia, 1 | AST harvest from in vitro cultures (size and age of cultures n/s) | ED | None | Following Wang et al. [32] | 0–9% survival | 2015 [99] | |
VI | None | Following Shatnawi et al. [83] | 0–1% survival | ||||
V. vinifera, 5 | AST and AxST harvested from in vitro cultures that are 2 weeks old (size n/s) | DV | Cultures on 1/2 MS + B5 vitamins ** + 20 g L−1 sucrose + 0.5 mg L−1 BA + 0.1 mM SA + 3 g L−1 gellan gum for 2 weeks at 24 °C | 1/2 MS + B5 vitamins ** with increased sucrose concentrations of 0.25 M, 0.5 M, 0.75 M and 1 M (every 24 h) + 3 g L−1 Gelrite (24 °C) | 2 M glycerol + 0.4 M sucrose (20 min at RT) → PVS2 (36–41.5 min at 0 °C) → LN | 13–30 | 2015 [62] |
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
V. vinifera, 3 | AST (1–2 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 10 days | VI | None | Medium with increased sucrose concentrations of 0.3, 0.5 and 0.75 M every 24 h (conditions n/s) | 2 M glycerol + 0.4 M sucrose (30 min at RT) → 2 M glycerol + 0.75 M sucrose (30 min at RT) → 1/2 PVS3 (30 min at RT) → 80 % PVS3 (60–90 min) | n/s after LN exposure | 2015 [100] |
V. vinifera, 4; V. riparia × V. rupestris, 1; V. vinifera Chasselas × V. berlandieri, 1 | AST and AxST (1–1.5 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | Nodal sections on 1/2 MS + B5 vitamins ** + 20 g L−1 sucrose + 0.5 mg L−1 BA + 0.1 Mm SA + 3 g L−1 gellan gum for 2 weeks at 24 °C | 1/2 MS + B5 vitamins ** with increased sucrose concentrations every 24 h of 0.25 M, 0.5 M, 0.75 M and 1 M + 3 g L−1 Gelrite (24 °C) | 2 M glycerol + 0.4 M sucrose (20 min at RT) → PVS2 (36–43 min at 0 °C) → LN | 7–45 | 2016 [16] |
V. vinifera, 6; V. pseudoreticulata, 2 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | None | 1/2 MS + 0.3 M sucrose + 0.16 mM GSH + 0.14 mM AsA + 7 g L−1 agar for 3 days at 24 °C | 2 M glycerol + 0.4 M sucrose (20 min at 24 °C) → 1/2 PVS2 (30 min at 0 °C) → PVS2 (50 min at 0 °C) → LN | 24–72 | 2018 [35] |
V. vinifera, 2; V. vinifera × V. labrusca, 1; V. pseudoreticulata, 1 | 43–59 | 2018 [51] | |||||
V. vinifera, 1; V. aestivalis, 1; V. afghanistan, 1; V. flexuosa, 1; V. palmate, 1; V. riparia, 1; V. rupestris, 1; V. sylvestris, 1; V. treleasii, 1 | AST (1–1.5 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 to 3 weeks | DV | Nodal sections on MS + 30 g L−1 sucrose + 0.2 mg L−1 BA + 0.1 mM SA + 1 mM GSH*** + 1 mM AsA + 3 g L−1 gellan gum for 2–3 weeks at 25 °C | 1/2 MS + 0.3 M sucrose + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (90 min at 0 °C) → LN | 25–43 | 2018 [36] |
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
V. champinii × 1613 Couderc, 1; V. berlandieri × V. riparia, 1; V. shampinii, 1 | ST (3 mm; type n/s) harvested from shoots of greenhouse plants | DV | None | 1/2 MS + 0.3 M sucrose for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 25 °C) → PVS2 (0–50 min at 0 °C) → LN | No shoot regrowth | 2019 [101] |
1/2 MS with increased sucrose concentrations every 24 h of 0.25, 0.5, 0.75 and 1 M (25 °C) | 2 M glycerol + 0.4 M sucrose (20 min at 25 °C) → PVS2 (50 min at 0 °C) → LN | 27–47 | |||||
V. vinifera, 2; V. berlandieri × V. riparia, 1 | AST (1 mm) harvested from the lateral shoots of nodal sections cultured for 2 weeks from growth chamber stock plants | DV | Nodal sections on MS + 30 g L−1 sucrose + 0.2 mg L−1 BA + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 1.5 % (v/v) PPM + 3 g L−1 gellan gum for 2 weeks at 25 °C | 1/2 MS + 0.3 M sucrose + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 1.5 % (v/v) PPM + 3 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (30–40 min at 0 °C) → LN | 43–64 | 2019 [102] |
V. vinifera, 2; V. actinifolia, 1; V. aestivalis, 1; V. jacquemontii, 1; V. flexuosa, 1; V. palmate, 1; V. riparia, 1; V. rupestris, 1; V. sylvestris, 1; V. ficifolia, 1; V. treleasi, 1; V. xnovae angeliae, 1 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | Nodal sections on MS + 30 g L−1 sucrose + 0.2 mg L−1 BA + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 2 weeks at 25 °C | 1/2 MS + 0.3 M sucrose + 0.1 Mm SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (90 min at 0 °C) → LN | 35–72 | 2019 [37] |
Pretreatment described above + cold-hardening for 2 weeks at 5 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (75 min at 0 °C) → LN | 43–70 | |||||
V. aestivalis, 1; V. jacquemontii, 1 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 weeks | DV | Nodal sections on MS + 30 g L−1 sucrose + 0.2 mg L−1 BA + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 2 weeks at 25 °C | 1/2 MS + 0.3 M sucrose + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (20 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (90 min at 0 °C) → LN | 53–70 | 2019 [76] |
V-CP | 2 M glycerol + 0.4 M sucrose (30 min at 22 °C) → PVS2 (40 min at 22 °C) → LN | 68–70 | |||||
Table 1 (continued) | |||||||
Species, No. Genotypes Tested | Explant Source | Cryo Method | Pretreatment | Preculture | Best Cryoprotectant/ Dehydration Treatment | Regrowth (%) | Year Ref. |
V. vinifera, 1 | AST (1 mm) harvested from the lateral shoots of in vitro nodal sections cultured for 2 to 3 weeks | DV | Nodal sections on MS + 30 g L−1 sucrose + 0.2 mg L−1 BA + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 2–3 weeks at 25 °C | 1/2 MS + 0.3 M sucrose + 0.1 mM SA + 1 mM GSH *** + 1 mM AsA + 3 g L−1 gellan gum for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (30 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (75 min at 0 °C) → LN | 68 | 2019 [103] |
AST (1 mm) harvested from growth chamber nodal sections cultured in vitro for 2 weeks before ST excision | Pretreatment described above + additional + 1.5% (v/v) PPM for 2 weeks at 25 °C | Preculture described above + additional + 1.5% (v/v) PPM for 3 days at 25 °C | 2 M glycerol + 0.4 M sucrose (30 min at 22 °C) → 1/2 PVS2 (30 min at 22 °C) → PVS2 (40 min at 0 °C) → LN | 43 | |||
V. vinifera, 1 | ST (size and type n/s) harvested from in vitro stock cultures that are 4 weeks old | ED | Cold-hardening for 4 weeks at 5 °C | 1/2 MS with increased sucrose concentrations every 24 h of 0.25, 0.5, 0.75 and 1 M + 2.5 g L−1 Phytagel (5 °C) | Bead desiccation to 18.4% → LN | 33% survival | 2021 [104] |
3. Pretreatment, Excision and Preculture of Shoot Tips
4. Methods for Shoot Tip Cryopreservation
4.1. Two-Step Cooling
4.2. Encapsulation-Dehydration
4.3. Vitrification
4.4. Encapsulation-Vitrification
4.5. Droplet-Vitrification
4.6. V Cryo-Plate
5. Shoot Tip Cryotherapy
5.1. Methods for Shoot Tip Cryotherapy
5.2. Mechanism Involved in Shoot Tip Cryotherapy for Eradication of Phloem-Limited Grapevine Viruses
5.3. Comparison of Virus Eradication Efficiency between the Traditional Methods and Shoot Tip Cryotherapy
6. Conclusions and Future Prospects
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
Abbreviations
AD | Apical dome |
AsA | Ascorbic acid |
AST | Apical shoot tips |
AxST | Axillary shoot tips |
BA | Benzylaminopurine |
DAS-ELISA | Double-antibody sandwich enzyme-linked immunosorbent assay |
DMSO | Dimethyl sulfoxide |
ELISA | Enzyme-linked immunosorbent assay |
FWB | Fresh weight basis |
IHC | Immunohistochemical |
LN | Liquid nitrogen |
LNV | Liquid nitrogen vapor |
LS | Loading solution |
ULS | Unloading solution |
SA | Salicylic acid |
GVA | Grapevine virus A |
GFKV | Grapevine fleck virus |
GFLV | Grapevine fanleaf virus |
GLRaV-1 | Grapevine leafroll-associated virus-1 |
GLRaV-2 | Grapevine leafroll-associated virus-2 |
GLRaV-3 | Grapevine leafroll-associated virus-3 |
GSH | Glutathione |
LP | Leaf primordia |
MD RT-PCR | Microtissue direct reverse transcription polymerase chain reaction |
MS | Murashige and Skoog (1962) medium |
NAA | Naphthaleneacetic acid |
PPV | Plum pox virus |
PVS | Plant vitrification solution |
PVS2 | Plant vitrification solution 2 |
PVS3 | Plant vitrification solution 3 |
RT-PCR | Reverse transcription polymerase chain reaction |
ROS | Reactive oxygen species |
ST | Shoot tips |
V cryo-plate | Vitrification cryo-plate |
ZR | Zeatin riboside |
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Specie and Genotype | Virus | Cryo Method | Explant | Regrowth (%) | Plant Conditions during Virus Detection | Virus-Free Frequency (%) | Virus Confirmation Method | Year Ref. |
---|---|---|---|---|---|---|---|---|
V. vinifera ‘Bruti’ | GVA | VI | ST (1 mm; type n/s) | 50 | Plants grown in the greenhouse for 4 months | 97 | Western blotting/ELISA | 2003 [64] |
ED | 62 | |||||||
V. vinifera ‘Black’ | GVA | ED | ST (1 mm; type n/s) | 59 | In vitro plants that are 2 to 4 months old | 42.2 | RT-PCR | 2011 [63] |
V. vinifera Nebbiolo | GVA GLRaV-3 | EV | AST (2 mm) | 15 | n/s | 100 | Multiplex RT-PCR | 2012 [167] |
V. vinifera Chardonnay | GFLV | DV | AST (1 mm) | 30.7 | In vitro plants that are 2 months old | 77.8 | ELISA | 2015 [34] |
V. vinifera Cabernet Sauvignon | GLRaV-3 | 41.6 | 100 | |||||
V. vinifera Pinot gris | GLRaV-2 | DV | AST and AxST (size n/s) | 13.6 | In vitro plants that are 3 months old/plants grown in the greenhouse for 3 and 6 months | 100 | DAS-ELISA | 2015 [62] |
V. vinifera Sauvignon blanc 316 | GLRaV-2 | 15.7 | 100 and 100 | |||||
V. vinifera Sauvignon blanc | GLRaV-1 and GLRaV-3 | 30 | 100 | |||||
V. vinifera ‘Lakemont Seedless’ | GLRaV-3 | 16.2 | 100 | |||||
V. vinifera Chardonnay | GLRaV-3 | 13 | 100 | |||||
V. vinifera Cabernet Sauvignon | GLRaV-3 | DV | AST (1 mm) | 59 | Plants grown in the screen-house for 12 months | 100 | RT-PCR and MD RT-PCR | 2018 [51] |
V. vinifera Chardonnay | 47 | |||||||
V. vinifera × V. labrusca ‘Kyoho’ | 51 | |||||||
V. pseudoreticulata ‘Hunan-1’ | 43 |
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Bettoni, J.C.; Marković, Z.; Bi, W.; Volk, G.M.; Matsumoto, T.; Wang, Q.-C. Grapevine Shoot Tip Cryopreservation and Cryotherapy: Secure Storage of Disease-Free Plants. Plants 2021, 10, 2190. https://doi.org/10.3390/plants10102190
Bettoni JC, Marković Z, Bi W, Volk GM, Matsumoto T, Wang Q-C. Grapevine Shoot Tip Cryopreservation and Cryotherapy: Secure Storage of Disease-Free Plants. Plants. 2021; 10(10):2190. https://doi.org/10.3390/plants10102190
Chicago/Turabian StyleBettoni, Jean Carlos, Zvjezdana Marković, Wenlu Bi, Gayle M. Volk, Toshikazu Matsumoto, and Qiao-Chun Wang. 2021. "Grapevine Shoot Tip Cryopreservation and Cryotherapy: Secure Storage of Disease-Free Plants" Plants 10, no. 10: 2190. https://doi.org/10.3390/plants10102190