A Comparison of Two Cryogenic Protocols in Cryopreservation of Apple Shoot Tips

Feng, Chungang; Li, B.Q.; Hu, Lingyun; Wang, M.R.; Wang, Q.C.

doi:10.17660/actahortic.2014.1039.20

Cited by 8 publications

(4 citation statements)

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“…The effects of droplet-vitri cation and encapsulation-dehydration procedures on post-cryotherapy recovery patterns in apple shoot tips have been compared [39,40]. These studies have shown that shoot tip regrowth rates obtained by droplet-vitri cation is more stable than rates yielded by encapsulationdehydration throughout the year, i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Feng et al [39] describe the procedure for droplet-vitri cation. First, shoot tips were cultured under stable conditions on a basic medium (BM) for 1 day, then transferred to the MS solution (0.8 M sucrose, 2 M glycerol), also for 1 day.…”

Section: Cryotherapymentioning

confidence: 99%

“…i Molecular marker (M); positive control (C+), apple stem pitting virus (ASPV)-infected apple cultivar 'Gala'; negative control (C-), virus-free 'Gala'; cultivars 'Qinyue' (1-3), 'Rui Xianghong' (4-7), 'Ruixue' (8)(9)(10)(11), and 'Ruiyang' (12)(13)(14)(15). ii, iii, and iv Molecular marker (M); positive control (C+), apple stem grooving virus (ASGV)-infected apple cultivar 'Gala'; negative control (C-), virus-free 'Gala'; cultivars 'Qinyue' (16)(17)(18)(19)(20)(21)(22)(23)(24)(25), 'Rui Xianghong' (26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36), 'Ruixue' (37)(38)(39)(40)(41)(42)(43)(44)(45)(46), and 'Ruiyang' (47)(48)(49)(50). v and vi Molecular marker (M); positive control (C+), apple chlorotic leaf spot virus (ACLSV)-infected apple cultivar 'Gala'; negative control (C-), virus-free 'Gala'; cultivars 'Qinyue' (51-54), 'Rui Xianghong' (54-58); 'Ruixue' (59-62), and 'Ruiyang' (62-65).…”

Section: Declarationsmentioning

confidence: 99%

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Combined thermotherapy and cryotherapy: an effective method for the eradication of three common viruses in cultivated apples

Liu¹,

Chen²,

Yan³

et al. 2020

Preprint

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Background Apple virus disease is a serious problem affecting apple quality and crop production efficiency, and solutions are being actively and urgently explored. In particular, the cultivation of virus-free seedlings is being increasingly explored as an important technical method for facilitating the virus-free breeding of excellent apple cultivars. In this study, the apple cultivars ‘Ruiyang’, ‘Ruixue’, ‘Rui Xianghong’, and ‘Qinyue’ were used as source materials to compare the effectiveness of thermotherapy and combined thermo- and cryotherapy in eradicating apple chlorotic leaf spot virus (ACLSV), apple stem grooving virus (ASGV), and apple stem pitting virus (ASPV). Results Viral presence in the in vitro shoots of the four apple cultivars was detected using a reverse transcription-polymerase chain reaction. All four cultivars were found to carry ASGV and ACLSV, and ‘Rui Xianghong’ and ‘Qinyue’ additionally carried ASPV. In vitro shoots of the four cultivars were subjected to varying periods of thermotherapy (0, 2, 4, 6, and 8 weeks). Shoot tips were then removed and half were cultured directly, and the remainder were cultured after cryotherapy treatment; their survival and regeneration rates were then evaluated to determine the efficiency of virus eradication after 8 months of subculture. We showed that the eradication of viruses in each of the cultivars and their survival rate are both the highest after 4 weeks of thermotherapy. When combined with cryotherapy, application of this period of thermotherapy achieved a 100% eradication of ASPV and ACLSV, and over 50% of ASGV. Conclusions This study demonstrated an effective methodology for developing both new virus-free plant materials and new apple cultivars that will support sustainability of the apple production industry in China and globally.

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Section: Discussionmentioning

confidence: 99%

Section: Cryotherapymentioning

confidence: 99%

Section: Declarationsmentioning

confidence: 99%

See 1 more Smart Citation

Combined thermotherapy and cryotherapy: an effective method for the eradication of three common viruses in cultivated apples

Liu¹,

Chen²,

Yan³

et al. 2020

Preprint

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“…It helps to reduce cost significantly and to improve the survival rate of explants due to a decrease in the effect of cryoprotectant usage and lower formation of ice structures [ 21 ]. Using the shoot tips is mostly preferable for woody plants [ 5 , 14 , 22 ], as helps to preserve the genotype of plants successfully and speeds up the process of culture recovery. Despite of many variants in the plant cryopreservation area, there is no common cryopreservation protocol suitable for all the woody plant cultures [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

Efficient Cryopreservation of Populus tremula by In Vitro-Grown Axillary Buds and Genetic Stability of Recovered Plants

Vidyagina

Kharchenko

Shestibratov

2021

Plants

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Axillary buds of in vitro microshoots were successfully frozen at –196 °C by the one-step freezing method using the protective vitrification solution 2 (PVS2). Microshoots were taken from 11 transgenic lines and three wild type lines. Influence of different explant pretreatments were analyzed from the point of their influence towards recovery after cryopreservation. It was found out that the use of axillary buds as explants after removal of the apical one increases recovery on average by 8%. The cultivation on growth medium of higher density insignificantly raises the regenerants survival rate. Pretreatment of the osmotic fluid (OF) shows the greatest influence on the survival rate. It leads to the increase in survival rate by 20%. The cryopreservation technology providing regenerants average survival rate of 83% was developed. It was based on the experimental results obtained with explant pretreatment. Incubation time in liquid nitrogen did not affect the explants survival rate after thawing. After six months cryostorage of samples their genetic variability was analyzed. Six variable simple sequence repeat (SSR) loci were used to analyze genotype variability after the freezing-thawing procedure. The microsatellite analysis showed the genetic status identity of plants after cryopreservation and of the original genotypes. The presence of the recombinant gene in the transgenic lines after cryostorage were confirmed so as the interclonal variation in the growth rate under greenhouse conditions. The developed technique is recommended for long-term storage of various breeding and genetically modified lines of aspen plants, as it provides a high percentage of explants survival with no changes in genotype.

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