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Abstracts: CryoLetters 22 (3), 2001

CryoLetters 22, 145-150 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

EFFECT OF cold EXPOSURE ON SURVIVAL AND STRESS PROTEIN EXPRESSION OF Drosophila melanogaster at DIFFERENT DEVELOPMENT STAGES

A.A. Tsutsayeva* and L.G. Sevryukova

Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of the Ukraine, Kharkov, 23 Pereyaslavskaya Str., Kharkov 61015, Ukraine

Summary

The effect of cold treatment (0C) on the survival of different developmental stages of the fruit-fly Drosophila melanogaster, and the dynamics of stress protein synthesis in the salivary gland were investigated. It was shown that low temperature stability of the D. melanogaster depended on both the stage of development and the duration of the low temperature influence. The synthesis of  new proteins was determined in the cells of the salivary glands of 5-day old larvae after cold treatment.  They were distinguished from the heat shock proteins with the exception of the protein with molecular weight 70 kDa.

Keywords: Protein synthesis, cold stress proteins, cold treatment, Drosophila melanogaster, heat shock proteins.

 

 

CryoLetters 22, 151-156 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

Cold hardiness in THE boreal adder, vipera berus

Stefan Andersson* and Lennart Johanssonç

*Animal ecology, Department of Ecology and Environmental Science, Umeň University, SE-901 87 Umeň, Sweden. çDepartment of Cellular and Developmental Biology, Umeň University, SE-901 87 Umeň, Sweden

Summary

In three freezing experiments we examined the freeze tolerance in newborn adders, Vipera berus, from East-central Sweden.  After a two to three hours exposure to freezing, ten out of eleven fast frozen snakes survived and recovered completely after being exposed to -3.1░C on average. In the other two experiments with fast and slow freezing followed by long exposures lasting for 22-30 hours, none of the snakes survived average exposures at -4.8░C or -3.8░C, respectively.  The glucose content of blood from frozen snakes was significantly higher than in unfrozen ones.  The increase was small and its contribution to freeze tolerance doubtful. Compared to other freeze tolerant reptiles, the adder was categorised as virtually non freeze tolerant capable of surviving only a short exposure not colder than approximately -4░C.  Supercooling could play a role in winter survival but their precise choice of hibernation site is probably the most important.

Keywords: Vipera berus, freeze tolerance, hibernation, cryoprotection, supercooling, boreal snakes

 

 

CryoLetters 22, 157-162 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

The  open pulled straw vitrification of ovine GV-oocytes: POSITIVE EFFECT OF RAPID cooling or RAPID thawing or both?

V. Isachenko1,2*, J.L. Alabart1, F. Nawroth2, E. Isachenko2, G. Vajta3 and J. Folch1

1Center for Agricultural Researches, Department of Animal Production, PO Box 727, Zaragoza, Spain; 2Department of Obstetrics and Gynecology; Cologne University, Kerpener-Str. 34, 50931 Cologne, Germany; 3Center for Early Human Development, Monash University, Clayton, Victoria 3168, Australia

Summary

Three protocols for the open pulled straw (OPS) vitrification of ovine GV-oocytes with slow cooling-rapid thawing, rapid cooling-slow thawing, and rapid cooling-rapid thawing were tested. The effect of ultra-rapid cooling in liquid nitrogen slush and superfine open pulled straws (SOPS) was also studied. Our results prove that both rapid cooling and rapid thawing are contributing in improved results achieved with the OPS technology. The use of liquid nitrogen slush is beneficial for ovine GV stage oocyte nitrification.

Keywords: ovine, oocyte, vitrification, regime, cooling, thawing.

 

 

CryoLetters 22, 163-174 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

Effects of Plant Growth Regulators on Survival and Recovery Growth following Cryopreservation

S.R. Turner1,2,* , D.H. Touchell3, T. Senaratna1, E. Bunn1, B. Tan2 and K.W. Dixon1

1. Kings Park and Botanic Garden, West Perth, WA, 6005, Australia.
2. Curtin University of Technology, Bentley, WA, 6102, Australia.
3. School of Forestry and Wood Products, Michigan Technological University, Townsend Drive, Houghton, Michigan, USA.
*Corresponding author; email: TURNERSR@POP.SES.CURTIN.EDU.AU

Summary

Studies on the effects of plant growth regulators (PGRs) on survival, recovery and post-recovery growth of shoot apices following cryopreservation are limited. In this study, the effects of plant growth regulators in both the culture phase and the recovery phase of cryostorage were examined for the rare plant species, Anigozanthos viridis ssp terraspectans Hopper. Survival of shoot apices was not correlated to cytokinin or auxin treatments administered in culture media prior to cryostorage. In recovery media, the plant growth regulators, kinetin, zeatin (cytokinins), IAA, (auxin) and GA3 were examined for their effect following cryopreservation. It was found that the application of a combination of cytokinin and 0.5 ÁM GA3 from day zero was the most appropriate for obtaining vigorously growing plantlets following LN immersion. This combination proved to be more effective than basal medium, zeatin or kinetin treatments.

Keywords: Anigozanthos viridis ssp terraspectans, plant growth regulators (PGR), cryopreservation, PVS2, vitrification, rare and endangered.

 

 

CryoLetters 22, 175-182 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

THE DUAL EFFECT OF ANTIFREEZE PROTEIN ON CRYOPRESERVATION OF RICE (Oryza sativa L.) EMBRYOGENIC SUSPENSION CELLS

Jun-Hui Wang*, Hong-Wu Bian, Yi-Xiang Zhang and Hai-Peng Cheng

College of Life Sciences, Zhejiang University, Hangzhou 310012, China
* Corresponding author, E-mail: junhuiwang@cls.zju.edu.cn

Summary

The effects of fish antifreeze protein AFP- on cryopreservation of rice suspension cells by three different protocols were investigated. During the two-step method, AFP- at 0.01 mg/ml significantly lowered the viability of both precultured and non-precultured cells. During the vitrification method, AFP- at 0.2 mg/ml improved the viability of suboptimally thawed cells; however, much higher doses of this protein (10mg/ml) attenuated the cell viability. During rapid freezing of rice cells in the solutions with relatively high (but non-vitrifying) concentrations of cryoprotectant, AFP- displayed protective action in the higher concentrated cryoprotectants and detrimental effect in more dilute ones. Taken together, it was concluded that, depending upon a number of factors discussed in the present paper, both positive effect and negative effect could be observed during application of AFP to cryopreservation of rice cells. The possible mechanism of this dual character was discussed.

Keywords: Antifreeze proteins, Cryopreservation, Vitrification, Rice, Embryogenic suspension cells

 

 

CryoLetters 22, 183-190 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

FREEZING TOLERANCE VERSUS FREEZING SUSCEPTIBILITY IN THE LAND SNAIL Helix aspersa (GASTROPODA: HELICIDAE)

Armelle Ansart1*, Philippe Vernon2 and Jacques Daguzan1

1UniversitÚ de Rennes 1, UMR 6553 Ecobio, bat.13, 263 Av. Gal Leclerc, CS 74205, 35042 Rennes Cedex, France
2UniversitÚ de Rennes 1, UMR 6553 Ecobio, Station Biologique, 35380 Paimpont, France armelle.ansart@univ-rennes1.fr

Summary

Freezing hardiness in ectotherms has often been separated into two categories, freezing tolerance and freezing susceptibility. But more complex classifications have also been proposed. Helix aspersa hibernates in Brittany during winter. It has a high temperature of crystallisation (between ľ1.2 and ľ7.4░C) and survives only few hours at freezing temperatures. Helix aspersa is a large snail, which needs a long time to freeze and can bear some ice formation in its tissues up to 60% of its total body water. It may be provisionally considered as a partially freezing tolerant species.

Keywords: cold hardiness, freezing hardiness, Gastropoda, Helix aspersa

 

 

CryoLetters 22, 191-198 (2001)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

ON PHASE TRANSITIONS IN THE WATER-ETHYLENE GLYCOL SYSTEM AT SUBZERO TEMPERATURES UNDER NON-ISOTHERMAL CONDITIONS

A.V.Zinchenko and V.D.Zinchenko*

Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of  Ukraine, Kharkov, 61015, 23, Pereyaslavskaya str., Kharkov, Ukraine, e-mail: cryo@online.kharkov.ua

Summary

The dynamic phase diagrams of the water-ethylene glycol (EG) system have been plotted at the warming rate of 0.4 C. min-1 after cooling of  samples down to -196C by quenching in liquid nitrogen or slow cooling at the rate of approximately 0.15C. min-1. The melting of two stable eutectics at the temperatures of -42.7C and -48.7C and the metastable eutectic at -63C was observed. Hydrates 1:1 crystallized separately from ice during warming both after quenching and after slow cooling. Glass devitrification and EG:3H2O hydrate melting took place during warming of the samples after quenching but these thermal features were absent after slow cooling in the EG concentration range of 42.0 - 63.5 weight percent. The notions about two-stage mechanism of ice nucleation in EG aqueous solutions might be critically revised.

Keywords: ethylene glycol, aqueous solutions, DSC, hydrates, phase transitions, glass transition

 

 

CryoLetters 22, 199-208
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU

THE USE OF MICROSATELLITE ANALYSIS IN Solanum tuberosum L. IN VITRO PLANTLETS DERIVED FROM CRYOPRESERVED GERMPLASM

Keith Harding1* and Erica E. Benson2

1DAMAR, Drum Road, Cuparmuir, Cupar, Fife, KY15 5RJ, Scotland. *Corresponding author. email: damar@tinyworld.co.uk.
2Plant Conservation Biotechnology Group, Division of Molecular and Life Sciences, School of Science and Engineering, University of Abertay Dundee, Bell Street, Dundee DD1 1HG Scotland. email: e.e.benson@abertay.ac.uk

Summary

This study reports the application of the encapsulation/dehydration cryopreservation and microsatellite analysis to the conservation of Solanum tuberosum cultivars Brodick and Golden Wonder. Cryopreserved shoot-tips were capable of up to 40% shoot and plantlet regeneration in Brodick and >60 % for Golden Wonder. Microsatellite analysis was used with genomic DNA of Golden Wonder and Desiree to establish DNA sequence length polymorphisms. As the basis of stability assessments this technique was applied to: (i) individual, field-grown, plants of Golden Wonder; (ii) individual Golden Wonder plants derived from a single parental tuber progeny; (iii) plantlets derived from in vitro cultures of Golden Wonder and Brodick and (iv) Golden Wonder and Brodick plantlets derived from cryopreserved germplasm.

Keywords: Solanum tuberosum, cryopreservation, microsatellites, genetic stability.

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