Cryopreservation of 12 Vitis Species Using Apical Shoot Tips Derived from Plants Grown In Vitro

Bettoni, Jean Carlos; Kretzschmar, Aike Anneliese; Bonnart, Remi; Shepherd, A.; Volk, Gayle M.

doi:10.21273/hortsci13958-19

Cited by 31 publications

(30 citation statements)

References 27 publications

(37 reference statements)

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“…With respect to previous cryogenic methodologies, the simplicity is among the main advantages of using DV, moreover the technique can be successfully replicated by the technical staff [ 23 ]. The V cryo-plate method appears to be very systematic and time saving [ 42 ].…”

Section: Discussionmentioning

confidence: 99%

“…Continuous research and technological evolution have markedly improved the cryogenic methodologies, allowing to enhance the recovery percentage of the species, as has occurred over the years, for example, in Vitis spp. [ 22 , 23 ] and potato [ 24 , 25 ]. The aim of this study was to assess the efficiency of the novel procedures, Droplet-vitrification and V cryo-plate, in order to improve the S. rebaudiana cryopreservation protocol.…”

Section: Introductionmentioning

confidence: 99%

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Two Advanced Cryogenic Procedures for Improving Stevia rebaudiana (Bertoni) Cryopreservation

Benelli̇

Carvalho

Merzougui

et al. 2021

Plants

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Cryopreservation is a useful tool for the long-term storage of plant genetic resources, and different cryogenic procedures have recently been developed. The present study focused on the use of the Droplet-vitrification (DV) and V cryo-plate protocol for the cryopreservation of Stevia rebaudiana in vitro-derived apical shoot tips and axillary shoot tips. A preliminary test showed that 90 and 120 min PVS2 (Plant Vitrification Solution 2) treatment significantly reduced the regrowth of the explants before immersion in liquid nitrogen (LN). For both procedures tested, the best osmoprotective condition for obtaining a higher regrowth of cryopreserved explants occurred when explants were PVS2 treated for 60 min. After direct immersion in LN, thawing and plating, the highest regrowth recorded was 80% with DV and 93% with V cryo-plate. Moreover, shoot tips proved to be a more suitable material for Stevia cryopreservation. A satisfactory vegetative regrowth was observed in the subcultures following cryopreservation by DV and V cryo-plate cryogenic procedures.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Two Advanced Cryogenic Procedures for Improving Stevia rebaudiana (Bertoni) Cryopreservation

Benelli̇

Carvalho

Merzougui

et al. 2021

Plants

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“…It was proved that pretreatment of explants is crucial in successful implementation of this method [ 4 ]. Using the most successful combinations made it possible to develop technology in aspen axillary bud cryopreservation with the average survival rate of 83%, which is a very high indicator [ 34 ]. Our studies demonstrated that the difference in regeneration for certain transgenic lines, such as transformants carrying the sp-Xeg gene, could be significantly lower in comparison with the others.…”

Section: Discussionmentioning

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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“…It is, therefore, important to establish minimum exposure time to cryoprotectant solutions in order to dehydrate tissue sufficiently yet avoid toxic effects (Azimi et al 2005). Chemicals such as DMSO, a frequent component in cryoprotectants, has been reported to be toxic to cells (Aronen et al 1999). The same is suggested for another component, glycerol (Wang et al 2014).…”

Section: Discussionmentioning

confidence: 99%

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

O’Brien

Hiti-Bandaralage

Folgado

et al. 2020

Plant Cell Tiss Organ Cult

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Cryopreservation combined with in vitro culture offers a safe and cost-effective method to conserve germplasm. Conservation of Persea spp. has been limited to heterozygous somatic embryos that are not true-to-type. A method for shoot-tip cryopreservation is vital to preserve the exact gene pool of interest. For the first time cryopreservation protocols for mature shoot tips of two avocado cultivars (cvs) 'Velvick' and 'Reed', were established. In vitro shoots were subjected to two different optimised pre-treatments; (1) cv 'Velvick'-high sucrose (0.3 M) or (2) cv 'Reed'-low temperature (10 °C) incubation, over a 2-week period prior shoot tip dissection. Two different plant vitrification solutions, plant vitrification solution 2 (PVS2) and vitrification solution L (VSL) were tested at 0 °C for 0, 10, 20, 30 and 40 min. Vitrified shoots were evaluated for survival and regrowth at 2 and 8 weeks after vitrification treatment and either with or without liquid nitrogen exposure. The study revealed that the optimal exposure time for each cultivar varied with the cryoprotectant used. After liquid nitrogen cv 'Velvick' highest regrowth levels were observed with 20 min exposure to either PVS2 or VSL, however, vigorous plants were produced only from VSL treated shoots. In the case of cv 'Reed' highest regrowth levels were observed with 10 min exposure to PVS2 however only morphologically normal plants were recovered from VSL treated shoots. Key messageCryopreservation of avocado shoot tips was successful using PVS2 and VSL with both recording similar recovery rates for 'Velvick' and 'Reed'; although only vigorous and morphologically normal plants were developed from VSL treatments.

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Cryopreservation of 12 Vitis Species Using Apical Shoot Tips Derived from Plants Grown In Vitro

Cited by 31 publications

References 27 publications

Two Advanced Cryogenic Procedures for Improving Stevia rebaudiana (Bertoni) Cryopreservation

Two Advanced Cryogenic Procedures for Improving Stevia rebaudiana (Bertoni) Cryopreservation

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

First report on cryopreservation of mature shoot tips of two avocado (Persea americana Mill.) rootstocks

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