scholarly journals Investigation of the post-cryogenic regeneration ability of potato varieties under different cultivation conditions

2019 ◽  
Vol 23 (3) ◽  
pp. 281-286 ◽  
Author(s):  
E. S. Bespalova ◽  
Yu. V. Ukhatova ◽  
N. N. Volkova ◽  
E. V. Oves ◽  
N. A. Gaitova ◽  
...  
Keyword(s):  
Shoot Tips ◽  
Axillary Buds ◽  
Regeneration Ability ◽  
Cultivation Conditions ◽  
Regeneration Rate ◽  
Dark Incubation ◽  
Potato Varieties ◽  
Droplet Vitrification ◽  
In Vitro Plants

Cryopreservation provides long-term storage of the gene pool of potato varieties in cryobanks at extremely low temperatures. Currently, droplet vitrification is the most widely used method for cryopreservation of potato varieties, which is constantly improving to increase the regeneration rates of the stored plant material. Different modifications of this method are used in the world’s leading potato genebanks. This paper presents the results of studying the effect of cultivation conditions after plunging into liquid nitrogen and thawing of shoots tips and axillary buds of in vitro plants on their postcryogenic recovery. The droplet-vitrification method modified at VIR was used for cryopreservation. The factor “prolonged dark incubation of explants” did not have a significant effect on the frequency of post-cryogenic regeneration of the studied varieties except for one variety (Krepysh), for which a significant increase in the regeneration rate was observed for the shoot tips cultivated in the darkness compared to the cultivation under the photoperiod 16/8 hours (light/darkness). The frequency of post-cryogenic regeneration of shoot tips was higher than that of the axillary buds for all varieties; however, these differences were significant (p < 0.05) only in two cases: for the variety Udacha (a photoperiod of 16/8 hours) and for the variety Krepysh (the dark incubation). The results of two-factor analysis of variance indicate that there is no effect of interaction of factor 1 (prolonged dark incubation) and factor 2 (explant type) on the ability of varieties to post-cryogenic recovery. Taking into account the obtained results, the further cryopreservation of an extended subset of 9 varieties was carried out using shoot tips, which, after freezing-thawing, were cultivated under the photoperiod of 16/8 hours. The frequency of post-cryogenic regeneration of these varieties varied from 30 to 60 %. A significant effect of genotype on postcryogenic recovery has been established. The ability of varieties to regenerate shoots after freezing and thawing was not related to the values of morphogenic indices of in vitro plants. The age of the meriklons (2–4 years) did not significantly affect either the morphogenic indices or the frequency of post-cryogenic regeneration.

Cryopreservation of Grapevine Shoot Tips from In Vitro Plants Using Droplet Vitrification and V Cryo-plate Techniques

2019 ◽  
pp. 469-482
Author(s):  
Jean Carlos Bettoni ◽  
Ranjith Pathirana ◽  
Remi Bonnart ◽  
Ashley Shepherd ◽  
Gayle Volk
Keyword(s):  
Shoot Tips ◽  
Droplet Vitrification ◽  
In Vitro Plants

scholarly journals In vitro Conservation and Cryopreservation of Nandina domestica, an Outdoor Ornamental Shrub

2013 ◽  
Vol 41 (2) ◽  
pp. 638 ◽  
Author(s):  
Aylin OZUDOGRU ◽  
Diogo Pedrosa Corrêa Da SILVA ◽  
Ergun KAYA ◽  
Giuliano DRADI ◽  
Renato PAIVA ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Storage Temperature ◽  
Sucrose Concentration ◽  
Aluminum Foil ◽  
Shoot Tips ◽  
In Vitro Conservation ◽  
Shoot Cultures ◽  
Storage Medium ◽  
Droplet Vitrification

The study focused on an economically-important ornamental outdoor shrub, Nandina domestica, with the aims to (i) optimize an effective in vitro conservation method, and (ii) develop a cryopreservation protocol for shoot tips by the PVS2 vitrification and droplet-vitrification techniques. For in vitro conservation of shoot cultures, the tested parameters were sucrose content in the storage medium (30, 45, 60 g/L) and storage temperature (4 °C or 8 °C). Cryopreservation was performed by applying the PVS2 vitrification solution, in 2-ml cryovials or in drops over aluminum foil strips, for 15, 30, 60 or 90 min at 0 °C, followed by the direct immersion in liquid nitrogen of shoot tips. Results show that N. domestica shoots can be conserved successfully for 6 months at both the temperatures tested, especially when 60 g/L sucrose is used in the storage medium. However, conservation at 4 °C showed to be more appropriate, as hyperhydricity was observed in post-conservation of shoots coming from storage at 8 °C. As for cryopreservation, a daily gradual increase of sucrose concentration (from 0.25 to 1.0 M) produced better protection to the samples that were stored in liquid nitrogen. Indeed, with this sucrose treatment method, a 30-min PVS2 incubation time was enough to produce, 60 days after thawing, the best recovery (47% and 50%) of shoot tips, cryopreserved with PVS2 vitrification and droplet-vitrification, respectively.

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

HortScience ◽  
2019 ◽  
Vol 54 (6) ◽  
pp. 976-981 ◽  
Author(s):  
Jean Carlos Bettoni ◽  
Aike Anneliese Kretzschmar ◽  
Remi Bonnart ◽  
Ashley Shepherd ◽  
Gayle M. Volk
Keyword(s):  
Cost Effective ◽  
Shoot Tips ◽  
Easy Access ◽  
Biotic And Abiotic Stresses ◽  
Breeding Programs ◽  
Vitis Species ◽  
Droplet Vitrification ◽  
Effective Manner ◽  
Ideal Method

The availability of and easy access to diverse Vitis species are prerequisites for advances in breeding programs. Plant genebanks usually maintain collections of Vitis taxa as field collections that are vulnerable to biotic and abiotic stresses. Cryopreservation has been considered an ideal method of preserving these collections as safety back-ups in a cost-effective manner. We report a droplet vitrification method used to cryopreserve 12 Vitis species (Vitis vinifera cvs. Chardonnay and ‘Riesling, V. actinifolia, V. aestivalis, V. jacquemontii, V. flexuosa, V. palmata, V. riparia, V. rupestris, V. sylvestris, V. ficifolia, V. treleasi, and V. ×novae angeliae) using shoot tips excised from plants grown in vitro. Our results demonstrated wide applicability of this technique, with regrowth levels at least 43% for 13 genotypes representing 12 Vitis species. We demonstrated that the droplet vitrification procedure can be successfully replicated by technical staff, thus suggesting that this method is ready for implementation.

scholarly journals Cryopreservation of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens) shoot tips by droplet-vitrification technique

2013 ◽  
Vol 21 (2) ◽  
pp. 79-85 ◽  
Author(s):  
Djurdjina Ružić ◽  
Tatjana Vujović ◽  
Radosav Cerović
Keyword(s):  
Liquid Medium ◽  
Room Temperature ◽  
Sucrose Concentration ◽  
Shoot Multiplication ◽  
Shoot Tips ◽  
Prunus Cerasus ◽  
Droplet Vitrification ◽  
Vitrification Solution ◽  
Gisela 5

ABSTRACT The droplet-vitrification technique was applied to in vitro shoot tips of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens). Explants were precultured in the dark at 23 °C, in liquid MS medium with a progressively increasing sucrose concentration (0.3 M for 15 h, then 0.7 M for 5 h). Loading involved a 30 min incubation of explants in a solution comprising 1.9 M glycerol and 0.5 M sucrose. Explants were dehydrated at room temperature using a solution PVS A3 [Murashige and Skoog (MS) liquid medium, 22.5% (w/v) sucrose, 37.5% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethylsulfoxide] for 30, 40 and 50 min and the PVS3 solution [MS liquid medium, 50% (w/v) sucrose, 50% (w/v) glycerol] for 60, 90 and 120 min. Explants were cooled by direct immersion in liquid nitrogen (LN) in 10 μl droplets of vitrification solution placed on aluminum foil strips. The foil strips were retrieved from LN and immersed in preheated (37 °C) unloading solution (0.8 M sucrose) for 30 s, and an equal volume of unloading solution at room temperature was added for further incubation for 30 min. Shoot tips were transferred onto the regrowth medium, cultivated in the dark for 7 days before being incubated under standard conditions. Three weeks after transferring the shoot tips onto the regrowth medium, the survival rate of control and cryopreserved explants of Gisela 5 dehydrated with PVS A3 was 100%, regardless of the treatment duration. After dehydration with solution PVS3, the survival varied between 70 and 100% for control explants and 78 and 95% for cryopreserved shoot tips. Gisela 5 shoot tips dehydrated for 40 min with PVS A3 vitrification solution demonstrated the best regrowth (38%). When using the PVS3 solution, survival of cryopreserved shoot tips was the highest (95%) after 60 min treatment followed by 40% regrowth. After three successive subcultures on shoot multiplication, medium shoots recovered viability, multiplication ability and morphology equal of that prior to cryopreservation.

scholarly journals IMPROVEMENT OF POTATO SEED MATERIAL AND ITS DIAGNOSTICS IN SYSTEM OF BIOTECHNOLOGY

2012 ◽  
Vol 14 ◽  
pp. 156-166
Author(s):  
T.M. Oleynik ◽  
K.A. Sloblodyan ◽  
S.A. Slobodyan ◽  
R.V. Gricay
Keyword(s):  
Molecular Diagnosis ◽  
Potato Virus ◽  
Antiviral Drugs ◽  
Field Conditions ◽  
Potato Seed ◽  
Potato Varieties ◽  
In Vitro Plants ◽  
Seed Material ◽  
Potato Virus M

The results of improvement studies of potato varieties by chemotherapy along with the use of antiviral drugs: RNA-ase, acyclovir, izatizon, and hydrochloride as well as data on the molecular diagnosis of X-and M-viruses in vitro plants, resulting from the recovery are presented. 3 lines free from virus X and 4 lines free from potato virus M were allocated. After the testing of variety changeability and its economically valuable characteristics in field conditions one of them will be selected and submitted to the Bank in vitro redeveloped varieties.

74. Cryopreservation of in vitro shoot tips of potato varieties by V- Cryo-plate method

Cryobiology ◽  
2012 ◽  
Vol 65 (3) ◽  
pp. 363 ◽  
Author(s):  
Shin-ichi Yamamoto ◽  
Kuniaki Fukui ◽  
Dai Hirai ◽  
Takao Niino
Keyword(s):  
Shoot Tips ◽  
Plate Method ◽  
Potato Varieties

scholarly journals Effect of the Duration of Liquid Nitrogen Storage on the Regrowth of Blackberry Cryopreserved by Droplet Vitrification

2017 ◽  
Vol 66 (1-2) ◽  
pp. 44-50
Author(s):  
Tatjana Vujović ◽  
Đurđina Ružić ◽  
Radosav Cerović
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Shoot Tips ◽  
Lower Survival Rate ◽  
Droplet Vitrification ◽  
Vitrification Solution ◽  
Long Term Preservation ◽  
Direct Immersion

SummaryIn vitro shoot tips of the blackberry cultivar ‘Čačanska Bestrna’ were cryopreserved using the droplet vitrification technique. Upon loading (30 min) in a solution of 1.9 M glycerol and 0.5 M sucrose, the explants were dehydrated for 40 min on ice with the PVS A3 vitrification solution (glycerol 37.5%, dimethyl sulfoxide 15%, ethylene glycol 15% and sucrose 22.5%) and for 40 min at room temperature with the PVS3 solution (glycerol 50% and sucrose 50%). They were subsequently frozen in individual microdroplets of vitrification solution, by direct immersion in liquid nitrogen (LN), and kept therein for 2, 4, 8 and 24 h. The explant rewarming was performed in an unloading solution (0.8 M sucrose) for 30 min at room temperature. The duration of LN exposure did not exert significant effects on the survival and regrowth of explants in both types of vitrification solutions. The survival and regrowth of cryopreserved shoot tips dehydrated with PVS3 solution ranged between 90–95% and 80–90%, respectively. However, dehydration with PVS A3 resulted in a lower survival rate (80–90%) and a considerably lower regrowth rate (55–65%) of explants. Monitoring the shoots regenerated in the in vitro culture revealed their normal capacity for multiplication and rooting in comparison with the controls, which fully confirms the purpose of cryopreservation in the long-term preservation of plant material.

scholarly journals 317 Establishment and Multiplication of in Vitro Pyrus spp. Cultivars

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 446E-447
Author(s):  
Adriana Cibele de Mesquita Dantas ◽  
Adriano Nunes Nesi ◽  
Lilia Bender Machado ◽  
Janny Haerter ◽  
Gerson Renan de Luces Fortes
Keyword(s):  
Tissue Culture ◽  
Shoot Tips ◽  
Axillary Buds ◽  
Temperate Climate ◽  
In Vitro Multiplication ◽  
Growth Room ◽  
Sodium Hypochloride

The culture of meristems, shoot tips, and axillary buds leads to the method of in vitro multiplication that is easily used and safe to obtain uniform copies with no undesirable variations. This work aimed to propagate five in vitro pear cultivars: Housui, Carrick, Nijisseiki, Packham's Triumph, and Red Bartlett. The work was carried out in the Tissue Culture Laboratory at Embrapa Temperate Climate. The plants were sprayed with benomyl (1.0 mg./L) and agrimicine (2.4 mg/L) in the fields, 2 weeks before the shoots were collected. The shoots were then cut with two buds with no leaves and desinfested with alcohol 70% for 10 s and 1% sodium hypochloride for 20 min, 50 explants, 25 buds, and 25 meristems, were then transferred to test tubes containing MS salts and vitamins, myo-inositol (100.0 mg/L), sucrose (30.0 g/L), agar (6.0 g/L), added to in mg/L: BAP (1.0), GA3 (0.1), and NAA (0.01). Three pear cultivars were used for in vitro multiplication (`Nijisseiki', `Red Bartlett', and `Housui') by using the same basal salt with N reduced to strength, added to (in mg/L): BAP (1.6), NAA (0.16). The material was kept in growth room under 16-h photoperiod, 25 ± 2 °C and 19 μMol·m-2·s-1 of flux radiation. The in vitro contaminations were mainly due to bacteria derived from the bud material (71.5%). Higher oxidation for meristem material was observed for `Carrick' and `Packham`s Triumph'. `Red Bartlett' showed the best results for all the variable studied, although all cultivars in general presented low response.

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