Grey poplar explant acclimation to improvethe dehydration tolerance and cryopreservation

Pokorná, Eva; Faltus, Miloš; Máchová, P.; Zámečnı́k, Jiřı́; Fulín, M.

doi:10.32615/bp.2019.148

Cited by 4 publications

(5 citation statements)

References 49 publications

(40 reference statements)

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“…However, axillary buds treated with PVS3 for 90 min and immersed, or not, in LN, showed higher regrowth when cold treatment was applied to the mother plant. These results, even if there was no significant overall effect on the explant regrowth, suggest that cold pre-treatment resulted in the different sensitivity of explants to cryoprotective solution, as previously observed in cryopreservation by grey poplar explant [65].…”

Section: Discussionsupporting

confidence: 78%

Cryopreservation of Hazelnut (Corylus avellana L.) Axillary Buds from In Vitro Shoots Using the Droplet Vitrification Method

Sgueglia

Frattarelli²,

Gentile³

et al. 2021

Horticulturae

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Cryopreservation by droplet vitrification was applied to hazelnut (Corylus avellana L.). axillary buds of the Italian cultivated variety Tonda Gentile Romana, which were collected from in vitro growing shoots, immersed in ice cooled PVS2 or PVS3 for 60 or 90 min, then transferred to a droplet of vitrification solution, placed on a strip of aluminium foil, and plunged into liquid nitrogen (LN). Additionally, the effect on the recovery of the mother plant after cryopreservation was evaluated, following a cold pre-treatment at 4 °C for 3 months. The highest regrowth percentage (56.7%) was obtained after applying PVS3 for 60 min, while the application of PVS2 for the same amount of time reduced regrowth to 41.5%. Increasing the exposure to vitrification solutions to 90 min reduced regrowth to 43.3% when PVS3 was applied, and 35.6% if PVS2 was used. The cold pre-treatment on the mother plant did not significantly improve overall regrowth. The cryopreservation process did not decline the rooting ability of the recovered shoots.

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Section: Discussionsupporting

confidence: 78%

Cryopreservation of Hazelnut (Corylus avellana L.) Axillary Buds from In Vitro Shoots Using the Droplet Vitrification Method

Sgueglia

Frattarelli²,

Gentile³

et al. 2021

Horticulturae

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“…Thermal analysis was performed using the Q2000 calorimeter (TA Instruments, New Castle, DE, USA) as previously described [67]. The analyzed solutions (see Additional file 2: Table S2 for a complete list) were loaded and hermetically sealed into aluminum pans.…”

Section: Differential Scanning Calorimetry (Dsc)mentioning

confidence: 99%

A mixture of innate cryoprotectants is key for freeze tolerance and cryopreservation of a drosophilid fly larva

Kučera

Moos

Štětina

et al. 2021

Preprint

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BackgroundOrganisms evolved biochemical strategies to cope with environmental stressors. For instance, insects that naturally tolerate internal freezing produce complex mixtures of multiple cryoprotectants (CPs). Better knowledge on composition of these mixtures, and on mechanisms of how the individual CPs interact, could inspire development of laboratory CP formulations optimized for cryopreservation of cells and other biological material.ResultsHere we identify and quantify (using high resolution mass spectrometry) a range of putative CPs in larval tissues of a subarctic fly, Chymomyza costata, that survives long-term cryopreservation in liquid nitrogen. The CPs (proline, trehalose, glutamine, asparagine, glycine betaine, glycerophosphoethanolamine, glycerophosphocholine, and sarcosine) accumulate in hemolymph in a ratio of 313:108:55:26:6:4:3:0.5 mmol.L-1. Using calorimetry, we show that the artificial mixtures, mimicking the concentrations of major CPs’ in hemolymph of freeze-tolerant larvae, suppress the melting point of water and significantly reduce the ice fraction. We demonstrate in a bioassay that mixtures of CPs administered through the diet act synergistically rather than additively to enable cryopreservation of otherwise freeze-sensitive larvae. Using MALDI-MSI, we show that during slow extracellular freezing of whole larvae trehalose becomes concentrated in partially dehydrated hemolymph and stimulates transition to the amorphous glass phase. In contrast, proline moves to the boundary between extracellular ice and dehydrated hemolymph and tissues where it likely forms a layer of dense viscoelastic liquid.ConclusionOur results suggest that different components of innate cryoprotective mixtures of freeze-tolerant insect act in synergy during extracellular freezing. We propose that transitions to amorphous glass (stimulated by trehalose) and viscoelastic liquids (having proline as major component) may protect macromolecules and cells from thermomechanical shocks associated with freezing and transfer into and out of liquid nitrogen.

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“…Despite of many variants in the plant cryopreservation area, there is no common cryopreservation protocol suitable for all the woody plant cultures [ 5 ]. It is connected with the peculiarities of culture: moist and size of tissue cells, physiological peculiarities, recovery rate and multiplication [ 5 , 21 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

“…That is why, work on creating cryopreservation protocols of the Populus genus plants lasts for many years, and significant success is achieved [ 4 , 14 , 15 , 23 ], but not all problems were solved so far. For in vitro plant material, a much larger than for plant material ex vitro variation in explant survival was observed [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

“…It is noted that the in vitro plant survival rate depended on the methods of explant pretreatment before immersion in liquid for cryopreservation [ 24 ]. It is proposed to make preliminary cultivation at low temperatures for plant explants pretreatment [ 14 , 15 , 23 ] and to add sugars as osmotic agents [ 14 , 23 ]. However, it was found out that conditions for pretreatment and storage should be selected individually for each specific genus and genotype [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

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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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Grey poplar explant acclimation to improvethe dehydration tolerance and cryopreservation

Cited by 4 publications

References 49 publications

Cryopreservation of Hazelnut (Corylus avellana L.) Axillary Buds from In Vitro Shoots Using the Droplet Vitrification Method

Cryopreservation of Hazelnut (Corylus avellana L.) Axillary Buds from In Vitro Shoots Using the Droplet Vitrification Method

A mixture of innate cryoprotectants is key for freeze tolerance and cryopreservation of a drosophilid fly larva

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

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