scholarly journals Morphophysiology and Cryopreservation of Seeds of Dendrobium Nobile Lindl. (Orchidaceae) at Different Stages of Development

2021 ◽  
Author(s):  
Galina L Kolomeitseva ◽  
Tatiana V Nikishina ◽  
Alexander V Babosha ◽  
Andrey S Ryabchenko ◽  
Olga N Vysotskaya
Keyword(s):  
Seed Viability ◽  
Embryo Sac ◽  
Dendrobium Nobile ◽  
Long Term Storage ◽  
Immature Seeds ◽  
Dendrobium Nobile Lindl ◽  
Term Storage ◽  
Pod Culture

Abstract We studied the seed maturation of Dendrobium nobile Lindl to establish the optimal stage for green pod culture and long-term storage in liquid nitrogen. It was found that the immature seeds can germinate in vitro starting from 3 months after pollination (MAP) but only develop up to the stage of embryo swelling or protocorm without rhizoids. The maximum staining of embryos with vital dyes FDA and TTC occurs at 4 and 5 MAP that corresponds to the release of suspensor beyond the embryo sac. Embryo staining does not correlate with germination capacity and seed viability in cryogenic storage. The immature seeds acquire resistance to freezing and drying in airflow at the age of 6 MAP. The time of technical maturity for green pod culture and cryopreservation may be reduced from 12 MAP to 6–7 MAP.

scholarly journals Barley Seeds miRNome Stability during Long-Term Storage and Aging

2021 ◽  
Vol 22 (9) ◽  
pp. 4315
Author(s):  
Marta Puchta ◽  
Jolanta Groszyk ◽  
Magdalena Małecka ◽  
Marek D. Koter ◽  
Maciej Niedzielski ◽  
...  
Keyword(s):  
Seed Viability ◽  
Small Rna Sequencing ◽  
The Novel ◽  
Seed Aging ◽  
Long Term Storage ◽  
Novel Mirna ◽  
Complex Biological Process ◽  
Almost All ◽  
Term Storage

Seed aging is a complex biological process that has been attracting scientists’ attention for many years. High-throughput small RNA sequencing was applied to examine microRNAs contribution in barley seeds senescence. Unique samples of seeds that, despite having the same genetic makeup, differed in viability after over 45 years of storage in a dry state were investigated. In total, 61 known and 81 novel miRNA were identified in dry seeds. The highest level of expression was found in four conserved miRNA families, i.e., miR159, miR156, miR166, and miR168. However, the most astonishing result was the lack of significant differences in the level of almost all miRNAs in seed samples with significantly different viability. This result reveals that miRNAs in dry seeds are extremely stable. This is also the first identified RNA fraction that is not deteriorating along with the loss of seed viability. Moreover, the novel miRNA hvu-new41, with higher expression in seeds with the lowest viability as detected by RT-qPCR, has the potential to become an indicator of the decreasing viability of seeds during storage in a dry state.

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 Development of cryopreservation methods of seed of Nepeta cataria

2021 ◽  
Vol 102 (2) ◽  
pp. 37-42
Author(s):  
Margarita Ishmuratova ◽  
◽  
Damirzhan Baigarayev ◽  
Saltanat Tleukenova ◽  
Elena Gavrilkova ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Seed Viability ◽  
Positive Temperature ◽  
Seed Rate ◽  
Nepeta Cataria ◽  
Long Term Storage ◽  
Medical Plant ◽  
Term Storage

This article presents the summarized data on cryopreservation of seeds of the medical plant Nepeta cataria. Cryopreservation is a highly promising method for saving of seed materials, allowing to organize long-term storage without viability loss. The purpose of present work is to optimize conditions of cryopreservation of seed materials of Nepeta cataria. Assessment of seed survival rate in the storage showed a linear decrease in seed viability and energy of germination. After 30 months of storage at the low positive temperature (+5 ºC) in paper pack seed rate decreased to 12.0 % and energy of germination to 11.2 %; after 4 years of storage seeds lost viability. During conduction of research the type of container, condition of thawing, optimal moisture of seeds and cryoprotectants are optimized. The optimal container for cryopreservation in liquid nitrogen was plastic cryo tubes; defrosting at room temperature. The best seed rate is found at moisture 3 %; the best cryoprotectant was glucose, the optimal concentration was 15 %. The result of the research is used for creation of the long-term storage medicinal cultures’ seed bank in the liquid nitrogen.

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 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.  

Long-term (35 years) cryopreservation of Echinococcus multilocularis metacestodes

Parasitology ◽  
2020 ◽  
Vol 147 (9) ◽  
pp. 1048-1054
Author(s):  
Teivi Laurimäe ◽  
Philipp A. Kronenberg ◽  
Cristian A. Alvarez Rojas ◽  
Theodor W. Ramp ◽  
Johannes Eckert ◽  
...  
Keyword(s):  
Echinococcus Multilocularis ◽  
Alveolar Echinococcosis ◽  
Geographic Origin ◽  
Experimental Animals ◽  
Long Term Storage ◽  
Term Maintenance ◽  
Term Storage

AbstractThe metacestode of Echinococcus multilocularis is the etiological agent of alveolar echinococcosis. The metacestode stage used for research is maintained in rodents by serial passages. In order to determine whether cryopreservation of E. multilocularis metacestodes would be suitable for long-term maintenance and replace serial passages, isolates of different geographic origin were cryopreserved in 1984–1986. The aim of the current study was to test the viability of cryopreserved isolates following long-term cryopreservation (up to 35 years) and to determine the phylogenetic clades these isolates belonged to. Cryopreserved isolates were tested for viability in vitro and in vivo in gerbils. In vitro results of 5 isolates indicated protoscolex survival in 13 of 17 experiments (76%) and metacestode survival in 5 of 12 (42%) in vivo experiments. In vivo results showed ‘abortive lesions’ in 13 of the 36 animals, 15 were negative and 8 harboured proliferating metacestode tissue containing protoscoleces. Genetic analysis confirmed the isolates belonged to European, Asian and North-American clades. In conclusion, the results of the current study indicate that metacestodes of E. multilocularis are able to survive long-term cryopreservation. Therefore, cryopreservation is a suitable method for long-term storage of E. multilocularis metacestode isolates and reduces the number of experimental animals.

Sign in / Sign up

Export Citation Format

Share Document