scholarly journals DEHIDRASI DAN PEMBEKUAN JARINGAN APEKS TEBU UNTUK PENYIMPANAN JANGKA PANJANG

2016 ◽  
Vol 21 (1) ◽  
pp. 25
Author(s):  
IKA ROOSTIKA ◽  
RARA PUSPITA DEWI LIMA WATI ◽  
DARDA EFENDI
Keyword(s):  
Liquid Nitrogen ◽  
Combined Treatment ◽  
Shoot Multiplication ◽  
Saccharum Officinarum ◽  
Freezing Method ◽  
Long Term Storage ◽  
Before And After ◽  
Freezing Liquid ◽  
Biology Group

<p>ABSTRAK<br />Tebu (Saccharum officinarum L.) merupakan tanaman yang<br />diperbanyak secara vegetatif. Kriopreservasi merupakan metode yang<br />paling sesuai untuk penyimpanan jangka panjang bagi tanaman yang<br />diperbanyak secara vegetatif. Dehidrasi dan pembekuan jaringan merupa-<br />kan tahapan paling kritis yang menentukan keberhasilan kriopreservasi.<br />Tujuan penelitian adalah untuk memperoleh durasi dehidrasi yang optimal<br />dan metode pembekuan jaringan apeks tebu. Penelitian dilakukan di<br />Laboratorium Kultur Jaringan, Kelompok Peneliti Biologi Sel dan<br />Jaringan, Balai Penelitian dan Pengembangan Bioteknologi dan Sumber-<br />daya Genetik Pertanian, Bogor pada Mei 2013 sampai Februari 2014.<br />Untuk optimasi metode dehidrasi, apeks direndam dalam larutan PVS2<br />(MS + gliserol 30% + etilen glikol 15% + dimetil sulfoksida 15% +<br />sukrosa 0,4 M) selama 10, 20, 30, dan 40 menit. Untuk optimasi metode<br />pembekuan, diujikan kombinasi perlakuan prakultur (dengan sukrosa 0;<br />0,1; dan 0,3 M selama 5 hari) dan pemuatan dalam larutan LS (MS +<br />gliserol 2 M + sukrosa 0,4 M) selama 0, 10, 20, dan 30 menit sebelum<br />tahapan dehidrasi dan pembekuan jaringan di dalam nitrogen cair (-<br />196 o C). Hasil penelitian menunjukkan durasi dehidrasi jaringan yang<br />terbaik adalah 30 menit dalam larutan PVS2. Kombinasi perlakuan<br />prakultur dengan sukrosa 0,3 M dan pemuatan dengan larutan LS selama<br />10 menit merupakan metode terbaik untuk pembekuan jaringan. Persentase<br />tumbuh sebelum dan setelah pembekuan dalam nitrogen cair berturut-turut<br />adalah 100 dan 40%. Setelah kriopreservasi, biakan mampu tumbuh<br />dengan tingkat multiplikasi tunas sekitar 10 tunas/eksplan. Metode yang<br />diperoleh pada penelitian ini berpeluang diterapkan untuk penyimpanan<br />plasma nutfah tebu dalam jangka panjang secara kriopreservasi dan<br />eliminasi patogen obligat secara krioterapi.<br />Kata kunci: Saccharum officinarum L., apeks, dehidrasi, pembekuan,<br />nitrogen cair</p><p>ABSTRACT<br />Sugarcane (Saccharum officinarum L.) is vegetatively propagated<br />plant. Cryopreservation is the most suitable method for long-term<br />preservation of vegetatively propagated plant. Dehydration and freezing<br />are critical steps of successful cryopreservation so that it should be<br />optimized. The research aimed to obtain the optimal duration of<br />dehydration and freezing method of sugarcane apex tissues. The<br />experiments were conducted at Tissue Culture Laboratory, Plant Cell<br />Tissue Biology Group, Indonesian Center for Agricultural Biotechnology<br />and  Genetic  Resources  Research  and  Development  on  May<br />2013−February 2014. To optimize dehydration method, the tissues were<br />exposured in PVS2 solution (MS + 30% glycerol + 15% ethylene glycol +<br />15% dimethyl sulphoxide + 0.4 M sucrose) for 10, 20, 30, and 40 minutes.<br />To optimize freezing method, the combined treatment of preculture with<br />sucrose (0, 0.1, dan 0.3 M) for 5 days and loading in LS solution (MS + 2<br />M glycerol + 0.4 M sucrose) for 0, 10, 20, dan 30 minutes) were tested<br />before dehydration for 30 minutes and freezing in liquid nitrogen (-196 o C).<br />The best duration of dehydration was 30 minutes. The combined treatment<br />of preculture on 0.3 M sucrose and loading for 10 minutes was the best<br />method for tissues freezing. Percentage of regrowth before and after<br />freezing in liquid nitrogen was 100 and 40% respectively. After<br />cryopreservation, the cultures could grow with high shoot multiplication<br />rate about 10 shoots/explant. The method resulted in this study can be<br />applied for long-term storage of sugarcane germplasms by cryopreser-<br />vation and (elimination of obligate pathogens by cryotherapy.<br />Keywords: Saccharum officinarum L., apex, dehydration, freezing, liquid<br />nitrogen.</p>

Evaluation of long-term frozen storage of plasma for measurement of high-density lipoprotein and its subfractions by precipitation

1990 ◽  
Vol 36 (5) ◽  
pp. 783-788 ◽  
Author(s):  
M N Nanjee ◽  
N E Miller
Keyword(s):  
High Density Lipoprotein ◽  
Liquid Nitrogen ◽  
Frozen Storage ◽  
Density Lipoprotein ◽  
High Density ◽  
Long Term Stability ◽  
Long Term Storage ◽  
Different Temperatures ◽  
Edta Plasma

Abstract The concentration of high-density lipoprotein cholesterol (HDL-C) in plasma is now established as an independent risk factor for coronary heart disease, but more data are needed on the relative risk-predictive powers of different HDL subclasses. For epidemiologic and clinical purposes, isolation of HDL from other lipoproteins and separation of its two major subclasses, HDL2 and HDL3, are performed most conveniently by precipitation. Although storage of plasma is commonly necessary, little information is available on the long-term stability of HDL subclasses at different temperatures. Therefore, we quantified HDL-C, HDL2-C, and HDL3-C by dual precipitation with heparin-MnCl2/15-kDa dextran sulfate (H-M/DS) in samples of EDTA-plasma from 93 healthy subjects, after storage for one to 433 days at -20 degrees C, at -70 degrees C, or in liquid nitrogen (-196 degrees C). Fourteen samples (15%) were stored for a year or longer. At -20 degrees C, HDL-C decreased by 4.8% per year and HDL3-C decreased by 6.9% per year (P = 0.002 for both variables) relative to results obtained with samples stored in liquid nitrogen; total cholesterol, HDL2-C, and triglyceride did not change significantly at this temperature. When stored at -70 degrees C, none of the lipids showed any change relative to results obtained with liquid nitrogen. Thus, long-term storage of EDTA-plasma at -20 degrees C is unsuitable for subsequent quantification of HDL-C and its subclasses by H-M/DS dual precipitation. Storage at -70 degrees C is preferable, and is as reliable as storage in liquid nitrogen.

scholarly journals EFFECT OF FREEZING ON THE MILK QUALITY OF COWS

2020 ◽  
pp. 54-57
Author(s):  
Lyubov Valentinovna Efimova ◽  
Tatyana Vyacheslavovna Zaznobina ◽  
Elena Vladimirovna Gatilova ◽  
Olga Valeryevna Ivanova
Keyword(s):  
Quality Indicators ◽  
Age Groups ◽  
Milk Quality ◽  
Long Term Storage ◽  
Before And After ◽  
Frozen State ◽  
Term Storage ◽  
Two Age Groups

Due to the fact that milk has a short shelf life, the issue of its freezing and long-term storage in the frozen state is relevant. A study was conducted on the effect of freezing on the milk quality after its defrosting of cows-mothers and cows-daughters of Red-Motley breed. As a result, it was established how milk quality indicators to change after five months of storage in a frozen state in cows of two age groups. A high correlation was established between quality indicators before and after freezing. The influence of the “freezing” factor on milk quality indicators was recognized statistically significant, the “generation” factor – not significant.

scholarly journals Does long-term storage of clay samples influence their mechanical characteristics?

2020 ◽  
Vol 57 (2) ◽  
pp. 304-309 ◽  
Author(s):  
Mustapha Abdellaziz ◽  
Mahmoud N. Hussien ◽  
Mohamed Chekired ◽  
Mourad Karray
Keyword(s):  
Mechanical Characteristics ◽  
Long Term Storage ◽  
Temperature And Humidity ◽  
Before And After ◽  
Prime Objective ◽  
Term Storage ◽  
Physical And Mechanical Characteristics

The prime objective of this study is to assess the influence of long-term storage on the physical and mechanical characteristics of clay samples. Samples from two different clays were sealed and stored in a temperature- and humidity-controlled room at the geotechnical laboratory of the Université de Sherbrooke for up to 27 years. The stored clay samples were tested before and after long-term storage and the results compared in this note. The comparison showed that even with long-term storage, the majority of the physical and mechanical characteristics of the samples were preserved.

Effect of storage temperature on spore viability and early gametophyte development of three vulnerable species of Alsophila (Cyatheaceae)

2010 ◽  
Vol 58 (2) ◽  
pp. 89 ◽  
Author(s):  
Y. Li ◽  
Y. L. Zhang ◽  
C. D. Jiang ◽  
T. Wang ◽  
Q. Wang ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Storage Temperature ◽  
Vulnerable Species ◽  
Spore Viability ◽  
Great Loss ◽  
Gametophyte Development ◽  
Long Term Storage ◽  
Duration Of Storage

To effectively preserve the vulnerable species of Alsophila, we studied the effects of varying the temperature and duration of storage on spore viability, early gametophyte development and the microstructure of brown spores of three Alsophila species. Spores of A. spinulosa (Wall. ex Hook.) Tryon and A. gigantea Wall. ex Hook. lost viability quickly when stored at room temperature and suffered from great loss when stored at –18°C from 6 to 12 months. Within 1 month, spore viability of A. spinulosa and A. gigantea stored at 4°C was higher than that of those stored in liquid nitrogen. In contrast, long-term storage in liquid nitrogen resulted in a comparatively small loss of viability for these two species. The spores of A. podophylla Hook. died within 3 months after storage at room temperature, 4°C and –18°C, and they died within 12 months when stored in liquid nitrogen. The spores of A. spinulosa and A. gigantea stored at room temperature, 4°C and –18°C, were prone to develop into abnormal gametophytes. These results suggest that storage of A. spinulosa and A. gigantea spores in liquid nitrogen is an effective method of preserving these vulnerable species. The reasons for the failure to preserve ephemeral A. podophylla spores by storage in liquid nitrogen are discussed.

Somatic polyembryogenesis in embryogenic cell masses of Piceaabies (Norway spruce) and Pinustaeda (loblolly pine) after thawing from liquid nitrogen

1987 ◽  
Vol 17 (9) ◽  
pp. 1130-1134 ◽  
Author(s):  
P. K. Gupta ◽  
D. J. Durzan ◽  
B.J. Finkle
Keyword(s):  
Liquid Nitrogen ◽  
Loblolly Pine ◽  
Somatic Embryos ◽  
Cell Mass ◽  
Embryogenic Cell ◽  
Long Term Storage ◽  
Embryogenic Cell Lines ◽  
Improvement Programs ◽  
Term Storage

We describe a method for the possible cryopreservation of embryogenic callus of Piceaabies and Pinustaeda at −196 °C and the regeneration of somatic embryos from thawed cells of subcultured embryonal–suspensor masses. Piceaabies and Pinustaeda were frozen without cryoprotective agent, in the presence of dimethyl sulfoxide (10%), or in a mixture of polyethylene glycol, glucose, and dimethylsulfoxide (10, 8, and 10% w/v, respectively). Cell masses placed in plastic vials or aluminum envelopes were frozen at 1 °C/min to −30 °C and then immersed for 10 min in liquid nitrogen. Cells were thawed rapidly and placed on modified MS subculture medium. Six to seven somatic embryos per gram of fresh weight were regenerated from each piece of frozen cell mass as compared with 12–13 embryos per gram from unfrozen cells. Post-thaw cell growth was inhibited initially by up to 5 weeks. Inhibition was reversed after the third 10-day subculture. Results suggest that the long-term storage of embryogenic cell lines in liquid nitrogen may be feasible for tree improvement programs in circumstances where testing of progeny may take several years.

scholarly journals Microbial occurrence in liquid nitrogen storage tanks: a challenge for cryobanking?

2021 ◽  
Author(s):  
Felizitas Bajerski ◽  
Manuela Nagel ◽  
Joerg Overmann
Keyword(s):  
Risk Assessment ◽  
Quality Management ◽  
Liquid Nitrogen ◽  
Potential Risk ◽  
Biological Materials ◽  
Cross Contamination ◽  
Storage Tanks ◽  
Long Term Storage ◽  
Term Storage

Abstract Modern biobanks maintain valuable living materials for medical diagnostics, reproduction medicine, and conservation purposes. To guarantee high quality during long-term storage and to avoid metabolic activities, cryostorage is often conducted in the N2 vapour phase or in liquid nitrogen (LN) at temperatures below − 150 °C. One potential risk of cryostorage is microbial cross contamination in the LN storage tanks. The current review summarises data on the occurrence of microorganisms that may compromise the safety and quality of biological materials during long-term storage. We assess the potential for the microbial contamination of LN in storage tanks holding different biological materials based on the detection by culture-based and molecular approaches. The samples themselves, the LN, the human microbiome, and the surrounding environment are possible routes of contamination and can cause cross contaminations via the LN phase. In general, the results showed that LN is typically not the source of major contaminations and only a few studies provided evidence for a risk of microbial cross contamination. So far, culture-based and culture-independent techniques detected only low amounts of microbial cells, indicating that cross contamination may occur at a very low frequency. To further minimise the potential risk of microbial cross contaminations, we recommend reducing the formation of ice crystals in cryotanks that can entrap environmental microorganisms and using sealed or second sample packing. A short survey demonstrated the awareness for microbial contaminations of storage containers among different culture collections. Although most participants consider the risk of cross contaminations in LN storage tanks as low, they prevent potential contaminations by using sealed devices and − 150 °C freezers. It is concluded that the overall risk for cross contaminations in biobanks is relatively low when following standard operating procedures (SOPs). We evaluated the potential sources in detail and summarised our results in a risk assessment spreadsheet which can be used for the quality management of biobanks. Key points • Identification of potential contaminants and their sources in LN storage tanks. • Recommendations to reduce this risk of LN storage tank contamination. • Development of a risk assessment spreadsheet to support quality management.

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.

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