scholarly journals The influence of disinfection methods and liquid phase media on Paphiopedilum insigne seeds germination and media supplements on morphological features of protocorms in tissue culture

2020 ◽  
Vol 19 (6) ◽  
pp. 7-14
Author(s):  
Monika Poniewozik ◽  
Marzena Parzymies ◽  
Paweł Szot
Keyword(s):  
Liquid Phase ◽  
Germination Rate ◽  
Casein Hydrolysate ◽  
Morphological Features ◽  
Cut Flower ◽  
Liquid Media ◽  
The Media ◽  
Whole Seed ◽  
Media Supplements

Paphiopedilum is a very original orchid, which labellum resembles a slipper. It is cultivated as a cut flower or a pot plant. The aim of the presented work was to estimate the influence of a disinfection method and addition of a liquid media on germination of Paphiopedilum insigne seeds in vitro. The whole seed capsules were disinfected with the use of 0.5% of AgNO3 for 20 min, 0.1% of HgCl2 for 5 s, 1% NaOCl for 30 min or immersed in a 96% ethanol and burned in a direct flame. The disinfected seeds were placed on a 1/4 MS (Murashige and Skoog) solidified media on top of which a liquid phase containing 1/4 MS elements, GA3 in concentration of 400 mg·dm–3 or distilled sterile water were added. The media without the liquid phase was also tested. It was noted that all methods of disinfection used effectively reduced contaminations of Paphiopedilum insigne seeds. The highest germination rate was observed when capsules were direct flamed and the liquid phase was added on top of the solidified media. The influence of growth regulators (BA, TDZ, KIN and 2,4-D) and casein hydrolysate added to the media on the morphological features of the obtained plants was also tested. It was observed that the most regenerated protocorms of the best quality were obtained when the media was supplemented with 1 mg·dm–3 of BA + 2 mg·dm–3 of TDZ or 5 mg·dm–3 of KIN + 1 mg·dm–3 of BA.

Urease activity related to the growth and differentiation of swarmer cells of Proteus mirabilis

1987 ◽  
Vol 33 (4) ◽  
pp. 300-303 ◽  
Author(s):  
Tianru Jin ◽  
R. G. E. Murray
Keyword(s):  
Proteus Mirabilis ◽  
Urease Activity ◽  
Casein Hydrolysate ◽  
Brain Heart Infusion ◽  
Liquid Media ◽  
Whole Cells ◽  
Maximum Proportion ◽  
The Media ◽  
Urease Production ◽  
Different Strains

Urease activity was measured using whole cells of both long (swarming) and short (nonswarming) populations of Proteus mirabilis from casein hydrolysate agar (CHA) and broth (CHB) cultures, and from brain heart infusion broth (BHIB) cultures. Urease is a constitutive enzyme for both long and short cells, but its activity was tremendously increased when urea was incorporated into the media. Urease production was also affected by culture age and media used. Before exponential phase, urease activity was very low, and it increased to its highest point after about 4 h in BHIB and 8 h in both CHA and CHB cultures at 37 °C. Long cells had higher urease activity than did short cells when grown on CHA, and was also expressed by two different strains cultured in BHIB. Strain PM23, in BHIB, was able to form long cells (swarming cells) to a maximum proportion after about 4 h, but strain IM47 could not differentiate in any of the liquid media. The former had more urease when swarming differentiation was initiated. PM23 grew relatively faster than IM47 when the former began to differentiate, but this fast growth could not be observed when nutrient broth or minimal medium was used. These observations suggest that long or swarming cells are "faster growing" rather than "nongrowing bactera."

scholarly journals An efficient regeneration system through in vitro somatic embryogenesis of barley (Hordeum vulgare L.)

2019 ◽  
Vol 27 ◽  
pp. 89-99
Author(s):  
M Haque ◽  
SMS Islam
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Casein Hydrolysate ◽  
Ms Medium ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Hordeum Vulgare L ◽  
The Media ◽  
Barley Genotypes

This study was carried out to improve an efficient protocol for in vitro callus induction and plant regeneration using Bangladeshi barley genotypes collected from BARI, Gazipur, Bangladesh. After sterilization embryos were separated carefully from mature seeds of six barley genotypes (BB-1, BB-2, BB-3, BB-4, BB-5 and BB-6) and cultured them in MS medium supplemented with various concentration and combination of PGRs for callus induction and regeneration. Out of six genotypes BB-6 showed highest (38.17%) callus induction in MS + 4.0 mg/l 2,4-D + 200 mg/l L-proline + 300 mg/l casein hydrolysate; whereas, BB-4 and BB-5 showed no callus induction in the same medium. For plant regeneration from embryogenic calli the same genotype (BB-6) also performed the best results (19.25%) in MS medium supplemented with 1.5 mg/l BAP + 30 g/l sucrose. Analysis of variance (ANOVA) showed highly significant differences among the media and the genotypes. J. bio-sci. 27: 89-99, 2019

In vitro germination of western larch pollen

2000 ◽  
Vol 30 (2) ◽  
pp. 329-332 ◽  
Author(s):  
Nicole Dumont-BéBoux ◽  
Bradley R Anholt ◽  
Patrick von Aderkas
Keyword(s):  
Pollen Grains ◽  
Major Effect ◽  
Basic Medium ◽  
Tube Length ◽  
In Vitro Germination ◽  
Liquid Media ◽  
Larix Occidentalis ◽  
Western Larch ◽  
The Media

We have been able to successfully germinate western larch (Larix occidentalis Nutt.) pollen in vitro. Pollen was rehydrated at 100% RH for 16 h before being sprinkled on semisolid and liquid media. The basic medium contained Brewbaker and Kwack minerals diluted 1:10 and was supplemented with polyethylene glycol 4000 and three different concentrations of sucrose. The flavonol quercetin was also included in half of the media. More pollen grains survived on liquid media, but semisolid media gave superior germination results. Two to 9% of the grains produced tubes. Quercetin had no major effect on germination, viability, or tube length.

scholarly journals Growing human trophoblasts in vitro: a review of the media commonly used in trophoblast cell culture

Reproduction ◽  
2020 ◽  
Vol 160 (6) ◽  
pp. R119-R128 ◽  
Author(s):  
Yohanes N S Nursalim ◽  
Cherie Blenkiron ◽  
Katie M Groom ◽  
Lawrence W Chamley
Keyword(s):  
Trophoblast Cell ◽  
Human Trophoblast ◽  
Vast Array ◽  
Trophoblast Stem ◽  
Gradual Development ◽  
The Media ◽  
Media Supplements ◽  
In The Mists ◽  
Trophoblast Culture

Trophoblasts are unique epithelial cells found only in the placenta. It has been possible to isolate and maintain human trophoblasts in in vitro culture for many decades. During this period there have been a vast array of media and supplements reported for trophoblast culture and often the reasons for using the media and specific supplements employed in any given laboratory have been lost in the ‘mists of time’. After a gradual development over many years this field has recently changed, with the publication of several reports of the isolation, growth and differentiation of human trophoblast stem or stem-like cells. This advance was made largely because of a greater understanding of the molecular pathways that control human trophoblasts and availability of media supplements that can be used to manipulate those pathways. We have searched the literature and here summarise many of the different media and supplements and describe how and why they were developed and are used to culture human trophoblasts.

Effects of tannic acid, ellagic acid, gallic acid and catechin on cellulose degradation by the rumen fungusneocallimastix frontalisstrain rel

1995 ◽  
Vol 1995 ◽  
pp. 147-147
Author(s):  
Samira Muhammed ◽  
Colin S. Stewart ◽  
Thomas Acamovic
Keyword(s):  
Gallic Acid ◽  
Tannic Acid ◽  
Ellagic Acid ◽  
Protein Hydrolysate ◽  
Cellulose Degradation ◽  
Rumen Fluid ◽  
Casein Hydrolysate ◽  
Nutritional Conditions ◽  
The Media

The ingestion of tannins and other polyphenols by ruminants can adversely affect the growth and fibre-digesting activities of the rumen microorganisms (Muhammedet al.1994). However, components of rumen liquor such as preformed monomers (amino acids, purines and pyrimidines) and other nutrients may protect the microorganisms, by providing nutritional conditions optimal for growth and energy metabolism. Rumen liquor also contains plant proteins, to which phenolic substances may bind preferentially. The influence of some polyphenols on the degradation of cellulose by the anaerobic fungusNeocallimastix frontalisstrain RE1 has been studied in two nutrient media considered to embrace the range of nutritional conditions that may occur in the rumen at different times. The media used were medium M2 of Hobson (1969) which contains rumen fluid casein hydrolysate and yeast extract, and the medium of Hungate and Stack (1982) (medium HS) which is chemically defined and in particular lacks protein hydrolysate and rumen fluid. Whatman no. 1 filter paper cellulose (30 mg) was the growth substrate. Filter-sterilized solutions of tannic acid (dissolved in dist. H20) or ellagic acid, gallic acid or catechin (dissolved in dimethylsulphoxide, DMSO) were added to the media (10 ml) prior to inoculation with strain REl (approx. 104zoospores) and incubation for 5 d at 38°C. Separate control incubations were performed with the 2 solvents (H20 and DMSO) used. The compounds tested were more inhibitory towards fungi grown in the defined medium (HS) than in the rumen fluid-containing medium M2 (Table 1). Thus tannic acid reduced cellulolysis to around 50 % of the relevant control values at a concentration of approx. 0.1 mM in HS, and 0.8 mM in M2. Corresponding values for ellagic acid were 0.15 mM and 0.5 mM, for gallic acid 0.25 mM and >3.6 mM and for catechin 0.15 mM and >2.4 mM respectively. It seems that these phenolic compounds are potent inhibitors of cellulolysis byNeocallimastixby mechanisms yet to be elucidated. It also appears that proteinaceous media reduce the inhibitory effects, suggesting that interactions occur between the proteins in the medium and the test compounds. It seems that in the rumen, dietary proteins and/or peptides may partly protect the fungi from the effects of polyphenolics as seen herein vitro.

scholarly journals Germination and Protocorm Formation of Ophrys Sphegodes Mill. – In Vitro Protocol for a Rare Orchid Species

2018 ◽  
Vol 67 (3-4) ◽  
pp. 196-201
Author(s):  
Jovana Dulić ◽  
Mirjana Ljubojević ◽  
Ines Prlainović ◽  
Goran Barać ◽  
Tijana Narandžić ◽  
...  
Keyword(s):  
Seed Germination ◽  
Germination Rate ◽  
Casein Hydrolysate ◽  
Habitat Destruction ◽  
Ex Situ ◽  
Organic Additives ◽  
Orchid Species ◽  
Short Period ◽  
Protocorm Formation

Summary Ophrys sphegodes Mill. is a wild orchid species which is threatened and protected due to its pollination biology, small seed and habitat destruction. The aim of this study was to establish asymbiotic germination protocol for the purpose of ex situ conservation. Two basal media Knudson C (KC) and Malmgren (MM), supplemented with organic additives (peptone (PE), L-glutamin (A)e, folic acid, casein hydrolysate (CA)) added separately and control media KC--C and MM--C were used in the present research. All the nutrition media contained 2% sucrose, 7% agar and 1% activated carbon, while their pH was adjusted to 5.8 ± 0.02 before autoclaving at 121 ° C for 20 minutes. The seeds were examined under two illumination conditions, 0/24 light/dark (L/D) and 16/8 L/D. The presented results indicate a huge influence of illumination and nutrition media on the seed germination and protocorm formation. The seed germination was overall significantly more successful in dark conditions (0/24 L/D) than with lighting (16/8 L/D). Protocorm, rhizoids and shoot formation were achieved only on the seeds cultured on MM medium, while the KC medium caused only swelling of the embryo. Organic additives had positive influence on the germination rate. According to the obtained results, the best germination rate and seedling development were achieved on MM-PE media, cultured in dark. The presented procedure accelerates the germination period and can provide a large number of plants in a relatively short period of time so it can be used for conservation programs and mass production protocol.

scholarly journals Targeting NSD2-mediated SRC-3 liquid–liquid phase separation sensitizes bortezomib treatment in multiple myeloma

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jing Liu ◽  
Ying Xie ◽  
Jing Guo ◽  
Xin Li ◽  
Jingjing Wang ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Drug Resistance ◽  
Phase Separation ◽  
Liquid Phase ◽  
Liquid Phase Separation ◽  
Acquired Drug Resistance ◽  
Bortezomib Treatment ◽  
Liquid Liquid Phase Separation

AbstractDevelopment of chemoresistance is the main reason for failure of clinical management of multiple myeloma (MM), but the genetic and epigenetic aberrations that interact to confer such chemoresistance remains unknown. In the present study, we find that high steroid receptor coactivator-3 (SRC-3) expression is correlated with relapse/refractory and poor outcomes in MM patients treated with bortezomib (BTZ)-based regimens. Furthermore, in immortalized cell lines, high SRC-3 enhances resistance to proteasome inhibitor (PI)-induced apoptosis. Overexpressed histone methyltransferase NSD2 in patients bearing a t(4;14) translocation or in BTZ-resistant MM cells coordinates elevated SRC-3 by enhancing its liquid–liquid phase separation to supranormally modify histone H3 lysine 36 dimethylation (H3K36me2) modifications on promoters of anti-apoptotic genes. Targeting SRC-3 or interference of its interactions with NSD2 using a newly developed inhibitor, SI-2, sensitizes BTZ treatment and overcomes drug resistance both in vitro and in vivo. Taken together, our findings elucidate a previously unrecognized orchestration of SRC-3 and NSD2 in acquired drug resistance of MM and suggest that SI-2 may be efficacious for overcoming drug resistance in MM patients.

scholarly journals G-quadruplex DNA inhibits unwinding activity but promotes liquid–liquid phase separation by the DEAD-box helicase Ded1p

2021 ◽  
Author(s):  
Jun Gao ◽  
Zhaofeng Gao ◽  
Andrea A. Putnam ◽  
Alicia K. Byrd ◽  
Sarah L. Venus ◽  
...  
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Liquid Phase Separation ◽  
Intrinsically Disordered ◽  
Dead Box ◽  
G4 Dna ◽  
G Quadruplex ◽  
The Dead ◽  
Liquid Liquid Phase Separation

G-quadruplex (G4) DNA inhibits RNA unwinding activity but promotes liquid–liquid phase separation of the DEAD-box helicase Ded1p in vitro and in cells. This highlights multifaceted effects of G4DNA on an enzyme with intrinsically disordered domains.

Development of an embryo germination protocol for shy-seeded grape (Vitis vinifera L.)

2021 ◽  
pp. 1-9
Author(s):  
Ashwini P. Benke ◽  
Ram Krishna ◽  
Roshni R. Samarth ◽  
Shweta S. Dhumal ◽  
Waquar A. Ansari ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Germination Rate ◽  
Germination Percentage ◽  
Vitis Vinifera L ◽  
Growth Media ◽  
Embryo Germination ◽  
Indole Butyric Acid ◽  
Grape Berries ◽  
Benzyl Amino Purine

Abstract Acquisition and germination of seeds are the most desired targets for the improvement of vegetatively propagated crops. In the present study, we developed a potential embryo germination protocol for the Red Globe grape cultivar having a low seed germination rate. Three grape berries at different developmental stages, viz. 50, 60 and 70 days after flowering (DAF), were selected for in-vitro embryo germination. Three growth media, namely Emershad and Ramming (ER), Nitsch and Nitsch (NN) and Murashige and Skoog (MS), and plant growth regulators (benzyl amino purine (BA), 0.5, 0.7 and 0.9 mg/l; indole butyric acid (IBA), 1.0, 1.5 and 2.0 mg/l; and gibberellic acid (GA), 0.1, 0.3 and 0.9 mg/l) were screened individually in different combinations with three amino acids, namely cysteine, glutamine and proline (2.0 μmol/l each). The maximum embryos germination percentage recorded at 70 DAF was 63.33, 47.78 and 45.56% in ER, NN and MS media, respectively, supplemented with 0.9 mg/l BA, 2.0 mg/l IBA, 0.9 mg/l GA and 2.0 μmol glutamine. Glutamine was found to have the most significant impact, and it improved the rescued embryos germination. The present study provides a potential recipe for a medium that can facilitate efficient germination of grape embryos.

Effect of microbial isolates on microbial yield estimation in RUSITEC system

1998 ◽  
Vol 22 ◽  
pp. 306-308
Author(s):  
M. D. Carro ◽  
E. L. Miller
Keyword(s):  
Protein Synthesis ◽  
Liquid Phase ◽  
Solid Phase ◽  
Liquid Fraction ◽  
Microbial Protein ◽  
Microbial Protein Synthesis ◽  
Continuous Culture System ◽  
The One

The estimation of rumen microbial protein synthesis is one of the main points in the nitrogen (N)-rationing systems for ruminants, as microbial protein provides proportionately 0.4 to 0.9 of amino acids entering the small intestine in ruminants receiving conventional diets (Russell et al., 1992). Methods of estimating microbial protein synthesis rely on marker techniques in which a particular microbial constituent is related to the microbial N content. Marker : N values have generally been established in mixed bacteria isolated from the liquid fraction of rumen digesta and it has been assumed that the same relationship holds in the total population leaving the rumen (Merry and McAllan, 1983). However, several studies have demonstrated differences in composition between solid-associated (SAB) and fluid-associated bacteria in vivo (Legay-Carmier and Bauchart, 1989) and in vitro (Molina Alcaide et al, 1996), as well in marker : N values (Pérez et al., 1996). This problem could be more pronounced in the in vitro semi-continuous culture system RUSITEC, in which there are three well defined components (a free liquid phase, a liquid phase associated with the solid phase and a solid phase), each one having associated microbial populations.The objective of this experiment was to investigate the effect of using different bacterial isolates (BI) on the estimation of microbial production of four different diets in RUSITEC (Czerkawski and Breckenridge, 1977), using (15NH4)2 SO4 as microbial marker, and to assess what effects any differences would have on the comparison of microbial protein synthesis between diets.This study was conducted in conjunction with an in vitro experiment described by Carro and Miller (1997). Two 14-day incubation trials were carried out with the rumen simulation technique RUSITEC (Czerkawski and Breckenridge, 1977). The general incubation procedure was the one described by Czerkawski and Breckenridge (1977) and more details about the procedures of this experiment are given elsewhere (Carro and Miller, 1997).

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