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 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 (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 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.

Patulin production by Penicillium expansum isolates from apples during different steps of long-term storage

2016 ◽  
Vol 9 (3) ◽  
pp. 379-388 ◽  
Author(s):  
N. De Clercq ◽  
G. Vlaemynck ◽  
E. Van Pamel ◽  
D. Colman ◽  
M. Heyndrickx ◽  
...  
Keyword(s):  
Phenotypic Diversity ◽  
Penicillium Expansum ◽  
Penicillium Species ◽  
Cold Temperatures ◽  
Long Term Storage ◽  
In Vitro Investigation ◽  
Duration Of Storage ◽  
Term Storage

Penicillium expansum is the principal cause of blue mould rot and associated production of patulin, a weak mycotoxin, in apples worldwide. P. expansum growth and patulin production is observed during improper or long-term storage of apples. We have investigated the extent to which each successive step during long-term storage contributes to patulin production in various P. expansum isolates. Fungal isolates collected on apples from several Belgian orchards/industries were identified to species level. Random amplification of polymorphic DNA (RAPD) analysis and β-tubulin gene sequencing identified P. expansum and Penicillium solitum as the most prevalent Penicillium species associated with Belgian apples. All 27 P. expansum isolates and eight reference strains were characterised for their patulin production capacity on apple puree agar medium for five days under classical constant temperature and atmosphere conditions. Under these conditions, a large range of patulin production levels was observed. Based on this phenotypic diversity, five P. expansum isolates and one reference strain were selected for in vitro investigation of patulin production under representative conditions in each step of long-term apple storage. Patulin accumulation seemed highly strain dependent and no significant differences between the storage steps were observed. The results also indicated that a high spore inoculum may lead to a strong patulin accumulation even at cold temperatures (1 °C) combined with controlled atmosphere (CA) (3% O2, 1% CO2), suggesting that future control strategies may benefit from considering the duration of storage under CA conditions as well as duration of deck storage.

scholarly journals Long-term in Vitro Regeneration of Hamelia patens

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873G-874
Author(s):  
D. Sankhla ◽  
T.D. Davis ◽  
N. Sankhla ◽  
A. Upadhyaya
Keyword(s):  
Culture Medium ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Prolific Shoot ◽  
Ornamental Shrub

This report describes an efficient in vitro regeneration protocol for H. patens (firebush), a heat-tolerant ornamental shrub native to tropical and subtropical America. Shoot cultures were initially established using shoot tips placed on MS-revised medium containing 2.3 μM 2,4-D, 2.3 μM kinetin, and 0.25% polyvinylpyrrolidone. Other types of explants (nodal and internodal segments, leaf pieces, floral buds) did not regenerate shoots when placed on this medium. Two-month-old plantlets derived from the shoot tips were subcultured on MS medium supplemented with 0.5 μM thidiazuron (TDZ), and within 3 to 4 weeks, some callus was produced at the root–shoot junction. When this callus, with a small portion of the root and shoots, was placed on MS medium with 0.05 μM TDZ and 0.01 μM ABA, prolific shoot formation occurred within 3 to 4 weeks followed by root formation. By regular subculturing every 5 to 6 weeks, hundreds of plantlets have been obtained over the past 3 years with no apparent decline in regeneration potential. Addition of activated charcoal (0.5%) to the culture medium has greatly improved growth of the plantlets.

scholarly journals Effect of Long-Term Storage on Mycobiota of Barley Grain and Malt

Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1655
Author(s):  
Soňa Felšöciová ◽  
Przemysław Łukasz Kowalczewski ◽  
Tomáš Krajčovič ◽  
Štefan Dráb ◽  
Miroslava Kačániová
Keyword(s):  
Barley Grain ◽  
In Vitro Assays ◽  
Production Cycle ◽  
Microscopic Fungi ◽  
Microbiome Composition ◽  
Long Term Storage ◽  
Fungal Microbiome ◽  
Term Storage

Contamination of malting barley grain and malt with micromycetes sampled at various periods post-harvest (3rd, 6th, and 9th month of storage) and types of storage (storage silo and floor warehouse) was investigated. Each of these barley grain samples was malted. This article reports on the changes in the fungal microbiome composition and their overall count in barley grain and malt. From the surface-disinfected barley grain samples collected immediately after harvest, there were eight genera isolated, with a predominance of Alternaria. A small increase of isolated microfungi was detected in barley stored in silo for 3 and 6 months (from 142 isolates to 149) and decreased below the number of isolates in barley before storage (133 isolates). Fungal count during storage gradually decreased up to 9 month in barley stored in floor warehouse (from 142 isolates to 84). The initial total count of microscopic fungi in malt before storage was the highest (112 isolates) with 7 genera detected, compared to malts prepared from barley stored for longer time (54 isolates, 7 genera, 9th month of storage). Alternaria was the most abundant and frequent genus. Quantitative representation of the filamentous microscopic fungi was lower compared to yeasts especially in barley and malt prepared from barley stored at third month of storage in both type of storage. Yeasts were identified from all grain samples and malt samples with mass spectrometry. Most attention was given to the widely distributed fungus Penicillium, 79% of strains produced at least one mycotoxin detected under in vitro assays using the TLC method (97% of them produced griseofulvin, 94% CPA, 79% patulin, 14% roquefortin C, and penitrem A was produced by two screening strains under laboratory conditions). It is therefore important to monitor the microflora throughout the production cycle of “barley to beer”.

scholarly journals Cryopreservation of Shoot Tips and Pollen of Potato

2017 ◽  
Vol 76 (1) ◽  
pp. 75-80
Author(s):  
Paulina Smyda-Dajmund
Keyword(s):  
Liquid Nitrogen ◽  
Shoot Tips ◽  
Popular Method ◽  
Long Term Storage ◽  
Term Storage ◽  
Direct Immersion

Abstract Cryopreservation is a frequently used method of long-term storage of potato meristems and pollen in liquid nitrogen (LN) in temperature of -196°C. This technique allows for theoretically unlimited storage of potato material. The most popular method of potato shoot tips preservation is cryopreservation by the solidification of liquids without crystallization (vitrification).The best method of pollen conservation is its direct immersion in LN. The successful regeneration after vitrification is genotype-dependent, which require optimization of protocol.

scholarly journals Long term storage and the compensatory growth of white shrimp Litopenaeus vannamei in aquaculture ponds

2017 ◽  
Vol 44 (3) ◽  
pp. 588-594
Author(s):  
Geraldo Fóes ◽  
Dariano Krummenauer ◽  
Gabriele Lara ◽  
Luis Poersch ◽  
Wilson Wasielesky Jr.
Keyword(s):  
Compensatory Growth ◽  
Management Practices ◽  
Specific Growth ◽  
Survival Rates ◽  
White Shrimp ◽  
Growth And Survival ◽  
Conventional Management ◽  
Long Term Storage ◽  
Term Storage

Effects of shrimp confinement in a situation of high density stocking in a long term nursery on their growth performance in grow out ponds. Were analized two nurseries with a density of 2000 shrimp m-2 were stocked at two different times. The first nursery (LTN) lasted 144 days, and the SGR of the animals was 3.0% day-1. The second nursery (STN) lasted 18 days and the specific growth rate (SGR) was 19.9% day-1. On the same day, shrimps were transferred to six lined ponds at a density of 20 shrimp m2 where they remained for 101 days. In the first biometry, the SGR in the LTN treatment, increased to 6.7% day-1 and in the STN it decreased to 5.0% day-1. At the end, shrimps of the LTN and STN treatments reached weights of 8.46 and 6.72 g and had productivities of 1287 and 1015 kg ha-1, respectively. Shrimps reared in nurseries for long periods showed growth and survival rates similar to those obtained using conventional management practices in grow out structures.

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