scholarly journals Cryopreservation of in vitro-grown shoot tips of chrysanthemum by encapsulation-dehydration

2013 ◽  
Vol 25 (2) ◽  
pp. 133-140 ◽  
Author(s):  
Małgorzata Zalewska ◽  
Dariusz Kulus
Keyword(s):  
Survival Rate ◽  
Sucrose Concentration ◽  
Multiple Shoots ◽  
Shoot Tips ◽  
Long Term Storage ◽  
The Media ◽  
And Storage ◽  
Cryopreservation Protocol

ABSTRACT Chrysanthemums are amongst the most economically important flowers in the world. The protection and storage of these valuable genetic resources is of great importance. Today, cryopreservation, or the storage of biological material at the temperature of liquid nitrogen (-196°C), is believed to be the most promising long-term storage method. To optimise the cryopreservation protocol, the shoot tips of Chrysanthemum × grandiflorum /Ramat./ Kitam. ‘Lady Orange’ and ‘Lady Salmon’ mutants were cryopreserved using the encapsulation-dehydration technique. During the experiment, the influence of sucrose concentration (2, 3 and 6%) during preculture and the concentration of kinetin (0.25, 0.5, 0.75 and 1.0 mg dm-3) in the regrowth medium were tested. A higher survival rate was observed for ‘Lady Salmon’. In general, the media with higher sucrose levels provided the best survival and recovery rates (35-40%). Kinetin had no influence on the survival rate; however, it influenced the morphogenesis of the plants. The lowest number of explants forming multiple shoots was observed on the medium with the lowest sucrose (during preculture) and kinetin (in the recovery medium) concentration. On the other hand, the best rhizogenesis efficiency was observed when 0.25 mg dm-3 kinetin was added. In conclusion, the composition of both preculture and recovery media need to be adjusted to single cultivars. The use of 3% sucrose (preculture) and 0.25 mg dm-3 kinetin (recovery) seems reasonable, since it guarantees a satisfying recovery rate of the explants and at the same time prevents the formation of callus and multiple shoots, stimulating the rooting instead.

scholarly journals Preservation of In Vitro Grown Shoot Tips of Potato (Solanum tuberosum L.) by Different Methods of Cryopreservation

1970 ◽  
Vol 10 ◽  
pp. 15-20
Author(s):  
Shambhu P. Dhital ◽  
Hira K. Manandhar ◽  
Hak T. Lim
Keyword(s):  
Sucrose Concentration ◽  
Shoot Tips ◽  
Efficient Tool ◽  
Long Term Storage ◽  
In Vitro Plantlets ◽  
Tuber Morphology ◽  
Vegetatively Propagated Plants ◽  
Term Storage

Cryopreservation has been recognized as a practical and efficient tool for long-term storage of vegetatively propagated plants. This study was conducted to investigate the effects of sucrose concentration, hardening temperature and different cryopreservation methods on the survival rate of potato shoot tips after cryopreservation. Excised shoot tips of in vitro plantlets of potato cultivars, Atlantic and Superior were cryopreserved by vitrification, encapsulationvitrification and encapsulation-dehydration. Cryopreservation by vitrification method was used to determine the optimum concentration of sucrose and cold hardening temperature during sub-culturing period to the donor plantlets. Nine-percent sucrose gave 46.7% survival in Atlantic and 40% in Superior. The most optimum hardening temperature for 50% survival in Atlantic and 43.3% in Superior was 10°C. In the case of comparative study of three different cryopreservation methods, the highest survival (52%) as well as regeneration (46%) were observed when the shoot tips were cryopreserved by encapsulation-vitrification method, and the lowest survival (36%) and regeneration (28%) from the vitrification. Plant and tuber morphology of potato regenerated after cryopreservation were similar to those of the non-cryopreserved in vitro plantlets (control). Thus, this study demonstrated that encapsulation-vitrification method was the most effective one among other methods for higher survival as well as regeneration in in vitro shoot tips of potato.Key words: Cryopreservation; Dehydration; Encapsulation; Potato; Regeneration; VitrificationDOI: 10.3126/njst.v10i0.2804Nepal Journal of Science and Technology Volume 10, 2009 December Page: 15-20

scholarly journals Cryopreservation of cherry rootstock Gisela 5 using vitrification procedure

2014 ◽  
Vol 41 (No. 2) ◽  
pp. 55-63 ◽  
Author(s):  
Dj. Ružić ◽  
T. Vujović ◽  
R. Cerović
Keyword(s):  
Sucrose Concentration ◽  
Survival Rates ◽  
Shoot Tips ◽  
Prunus Cerasus ◽  
Ms Medium ◽  
Long Term Storage ◽  
Term Storage ◽  
Gisela 5

In vitro-grown shoot tips of Gisela 5 (Prunus cerasus × Prunus canescens) cherry rootstock were tested for regrowth after cryopreservation using vitrification technique. Explants were precultured in the dark at 23°C, in a liquid MS medium with a progressively increasing sucrose concentration (0.3 M for 15 h, then 0.7 M for 5 h), and subsequently loaded in a solution containing 2 M glycerol and 0.4 M sucrose for 20 minutes. Shoot tips were dehydrated at 0°C using either the original PVS2 or modified PVS2 solution (PVS A3 – 22.5% sucrose, 37.5% glycerol, 15% ethylene glycol and 15% DMSO) for 30, 40 and 50 minutes. The survival and regrowth of the cryopreserved shoot tips dehydrated with the original PVS2 solution ranged between 36–54% and 8–17%, respectively. However, the dehydration with the PVS A3 solution resulted in considerably higher survival rates (81–92%), as well as higher regrowth rates (39–56%) after cryopreservation. These results prove the feasibility of the PVS A3-based vitrification technique for a long-term storage of this genotype.  

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

Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 77
Author(s):  
Elena O. Vidyagina ◽  
Nikolay N. Kharchenko ◽  
Konstantin A. Shestibratov
Keyword(s):  
Survival Rate ◽  
Axillary Buds ◽  
Long Term Storage ◽  
Average Survival ◽  
Transgenic Lines ◽  
Ssr Loci ◽  
One Step ◽  
The One

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.

scholarly journals A modified droplet vitrification method for cryopreservation of shoot tips from in vitro potato plants

2019 ◽  
Vol 23 (4) ◽  
pp. 422-429 ◽  
Author(s):  
T. A. Gavrilenko ◽  
N. A. Shvachko ◽  
N. N. Volkova ◽  
Yu. V. Ukhatova
Keyword(s):  
Plant Genetic Resources ◽  
Original Method ◽  
Shoot Tips ◽  
Droplet Vitrification ◽  
Modified Method ◽  
Long Term Storage ◽  
Common Potato ◽  
The Impact

Collections of common potato maintained in the field genebanks suffer significant losses due to the impact of extreme environmental factors, diseases and pests. The solution of the problem of safe long-term preservation of common potato accessions is to create doublet in vitro and cryo-collections. Cryogenic collections are stored at ultra-low temperatures in cryobanks. Several methods of potato cryoconservation are known, of which the droplet vitrification method developed by B. Panis with colleagues in 2005 is the most widely used in genebanks. This paper provides a detailed description of the modified method of droplet vitrification, which is used for cryopreservation of apexes (shoot tips) of potato in vitro plants at the N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR). The method modified at VIR includes the main steps of the original droplet-vitrification method developed by B. Panis and colleagues: 1) preparation of plant material, 2) isolation of shoot tips, 3) treatment of explants with cryoprotector solutions, 4) freezing/immersion in liquid nitrogen, 5) thawing, 6) post-cryogenic recovery and evaluation of viability and regeneration capacity. The modifications of stages 1, 2 and 6 proposed at VIR lead to a significant reduction in the duration of cryopreservation experiments in comparison with the original method of B. Panis. This paper presents the results of cryopreservation of modern potato cultivars and South American landraces which were obtained using the method of droplet vitrification as modified at VIR. The majority (76.7 %) of the studied accessions of cultivated potato were characterized by high rates of postcryogenic recovery (40–95 %) and 23.3 % of the samples had the values of postcryogenic regeneration from 20 to 39 %, which corresponds to the minimal permissible values for long-term storage in a cryobank. Currently the modified droplet-vitrification method is used for further expanding of the VIR potato cryocollection.

scholarly journals Application of vitrification-derived cryotechniques for long-term storage of poplar and aspen (Populus spp.) germplasm

2008 ◽  
Vol 18 (2) ◽  
pp. 160 ◽  
Author(s):  
I. TSVETKOV ◽  
C. BENELLI ◽  
M. CAPUANA
Keyword(s):  
Woody Species ◽  
Growth Conditions ◽  
Shoot Tips ◽  
Hybrid Aspen ◽  
White Poplar ◽  
Long Term Storage ◽  
Apical Buds ◽  
High Level

The application of three different vitrification-based freezing strategies for the cryostorage of white poplar (Populus alba L.) and hybrid aspen (P. tremula L. × P. tremuloides Michx.) have been assessed. The PVS2 vitrification protocol was successfully applied to two white poplar in vitro clones stored for more than 6 months in slow-growth conditions (4 °C, in darkness) and showing clear signs of explant etiolation and decay. After 60 min of PVS2 treatment, P. alba L. (cv. Villafranca) explants isolated from axillary buds demonstrated significantly better potential for post-freeze regrowth (64%) compared to those obtained from apical buds (17%). Similarly, a high level of survival (78%) of the frozen hybrid aspen shoot tips was recorded following the application of the same technique. Using the ‘encapsulation-vitrification’ procedure, no toxic effects of the PVS2 treatment were noticed after 120 min exposure, however none of the cryopreserved (poplar and aspen) explants survived after 3 weeks. In contrast, the ‘droplet-vitrification’ technique appeared to be very efficient in the cryopreservation of white poplar shoot tips, which increases the opportunities for wider application of this method in other woody species.;

Pemanfaatan Kriopreservasi untuk Penyimpanan Bibit Anggrek Jangka Panjang

2020 ◽  
Vol 10 (2) ◽  
pp. 71
Author(s):  
Arkan Setiaji ◽  
Rr Rifka Annisa
Keyword(s):  
Review Article ◽  
Breeding Programs ◽  
Cold Temperatures ◽  
Orchid Species ◽  
Long Term Storage ◽  
Cryopreservation Techniques ◽  
Cryopreservation Protocol ◽  
Term Storage

ABSTRAKTanaman anggrek (Orchidaceae) telah menjadi komoditas penting di pasar perdagangan internasional karena keeksotisan dan daya tahan bunganya. Meskipun keragaman anggrek spesies menduduki peringkat kedua setelah Asteraceae dan hibridanya menduduki peringkat pertama, namun anggrek menghadapi ancaman kepunahan dan penurunan kualitas genetik. Program pemuliaan, seperti persilangan dan penyisipan gen unggul, serta perbanyakan in vitro perlu didukung dengan cara memastikan ketersediaan eksplan. Dalam rangka penyimpanan untuk kebutuhan jangka panjang, eksplan dapat disimpan menggunakan teknik kriopreservasi. Review ini bertujuan untuk memberikan gambaran umum terhadap kemajuan penelitian di bidang kriopreservasi, khususnya tanaman anggrek. Melalui program penyimpanan jangka panjang, gen-gen dan eksplan unggulan baik untuk pengembangan anggrek hibrida atau khususnya untuk tujuan konservasi anggrek langka bisa dipastikan ketersediaannya. Dalam artikel review ini telah dibahas berbagai riset tentang teknik kriopreservasi pada biji, protokorm, dan serbuk sari anggrek. Kemajuan pengembangan teknik telah dicapai, namun masih diperlukan banyak modifikasi agar suatu protokol kriopreservasi dapat diterapkan pada semua spesies dan hibrida anggrek.Kata kunci: kriopreservasi; anggrek; suhu dingin; eksplan ABSTRACTOrchid plants (Orchidaceae) have become important commodities in the international trades market because of their exoticism and flower endurance. Although the diversity of orchid species is ranked second after Asteraceae and number of hybrids are ranked first, but orchids face the threat of extinction and genetic degradation. Breeding programs, including such as crossing and insertion of superior genes, and in vitro propagation need to be supported by ensuring the availability of explants. In the context of maintaining storage for long-term needs, explants can be stored using cryopreservation techniques. This review aims to provide an overview of the progress of research in the field of cryopreservation, specifically orchid plants. Through a long-term storage program, superior genes and explants both for the development of hybrid orchids or in particular for the purpose of conservation of rare orchids can be ensured availability. In this review article various researches on cryopreservation techniques in seeds, protocorms, and pollen have been discussed in this article. Advances in technical development have been achieved, but many modifications are still needed so that a cryopreservation protocol can be applied to all orchid species and hybrids.Keywords: cryopreservation; orchids; cold temperatures; explants

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

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 277
Author(s):  
Carla Benelli ◽  
Lara S. O. Carvalho ◽  
Soumaya EL merzougui ◽  
Raffaella Petruccelli
Keyword(s):  
Plant Genetic Resources ◽  
Stevia Rebaudiana ◽  
Preliminary Test ◽  
Shoot Tips ◽  
Axillary Shoot ◽  
Suitable Material ◽  
Long Term Storage ◽  
Stevia Rebaudiana Bertoni

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.

MODERN PROBLEMATICS AND ANALYSIS OF EFFICIENCY OF APPLICATION OF DIFFERENT APPROACHES TO DESTRUCTION OF INFORMATION ON MAGNETIC MEDIA

2019 ◽  
Vol 2 (12) ◽  
pp. 154-164
Author(s):  
O. Semenenko ◽  
O. Vodchyts ◽  
V. Koverga ◽  
R. Lukash ◽  
O. Lutsenko
Keyword(s):  
Armed Forces ◽  
Information Storage ◽  
Storage Device ◽  
Magnetic Media ◽  
Long Term Storage ◽  
Physical Impact ◽  
And Storage ◽  
Term Storage

The introduction and active use of information transmission and storage systems in the Ministry of Defense (MoD) of Ukraine form the need to develop ways of guaranteed removal of data from media after their use or long-term storage. Such a task is an essential component of the functioning of any information security system. The article analyzes the problems of guaranteed destruction of information on magnetic media. An overview of approaches to the guaranteed destruction of information on magnetic media of different types is presented, and partial estimates of the effectiveness of their application are given by some generally accepted indicators of performance evaluation. The article also describes the classification of methods of destruction of information depending on the influence on its medium. The results of the analysis revealed the main problems of application of software methods and methods of demagnetization of the information carrier. The issue of guaranteed destruction of information from modern SSD devices, which are actively used in the formation of new systems of information accumulation and processing, became particularly relevant in the article. In today's conditions of development of the Armed Forces of Ukraine, methods of mechanical and thermal destruction are more commonly used today. In the medium term, the vector of the use of information elimination methods will change towards the methods of physical impact by the pulsed magnetic field and the software methods that allow to store the information storage device, but this today requires specialists to develop new ways of protecting information in order to avoid its leakage.

scholarly journals In vitro Propagation of Polygonum hydropiper L. from Shoot Tips

2010 ◽  
Vol 20 (1) ◽  
pp. 73-79 ◽  
Author(s):  
M. F. Hasan ◽  
B. Sikdar
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Root Induction ◽  
Shoot Induction ◽  
Multiple Shoot Induction ◽  
Polygonum Hydropiper

An efficient protocol for plant regeneration through multiple shoots induction from shoot tips of Polygonum hydropiper (L.) was established. The highest percentage (96.6) of multiple shoot induction and number of shoots (9.0) per culture were found on MS supplemented with 2.0 mg/l Kn. The induced shoots were excised and inoculated on to MS contains different concentrations of IBA or NAA for rooting. The highest percentage (90.0) of root induction and the highest number of roots per shoot (12.0) was found on MS having 1.0 mg/l IBA. Well rooted plantlets were acclimated properly and transplanted in the soil under natural condition, where cent per cent plantlets survived and grew successfully. Key words:  Polygonum hydropiper, Shoot tips, In vitro propagation D.O.I. 10.3329/ptcb.v20i1.5970 Plant Tissue Cult. & Biotech. 20(1): 73-79, 2010 (June)

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