Phytagel

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Japan, Canada, India, Netherlands, Switzerland, France

Phytagel is a natural polysaccharide derived from Sphingomonas paucimobilis bacteria. It is a versatile gelling agent used in various laboratory applications, including cell and tissue culture, microbiology, and biochemistry. Phytagel forms clear, stable gels that can withstand a wide range of pH, temperature, and ionic conditions.

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Lab products found in correlation

Sucrose, by Merck Group (13 mentions) Tween 20, by Merck Group (7 mentions) Myo inositol, by Merck Group (4 mentions) Ms basal medium, by Merck Group (3 mentions) Plant preservative mixture, by Nacalai Tesque (3 mentions) 2 morpholinoethanesulfonic acid mes monohydrate, by Nacalai Tesque (3 mentions) Gamborg vitamin, by Merck Group (3 mentions) Cu 36l5, by Percival (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Sunshine mix la4, by Sun Gro Horticulture (2 mentions) Plant agar, by Duchefa Biochemie (2 mentions) Insulin pen needle 34 g, by Terumo (2 mentions) Murashige and skoog ms medium, by Duchefa Biochemie (2 mentions) A1049, by Merck Group (2 mentions) Gellan gum, by Merck Group (2 mentions) Activated charcoal, by Merck Group (2 mentions) Murashige and skoog basal medium, by Merck Group (2 mentions) Msp01, by Caisson (2 mentions) Sodium hypochlorite, by Fujifilm (2 mentions) Phytagel c, by Merck Group (2 mentions)

Close spelling variants of this product

Phytagel agar (4 citations) Gellan gum (phytagel, (2 citations) Phytagel powder (2 citations) L-1 Phytagel (2 citations) Phytagel medium (2 citations) Phytagel (P8169; (2 citations) Phytagel C (2 citations)

237 protocols using phytagel

Callus Maturation and Somatic Embryogenesis

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For the maturation treatment, three colonies containing 200 mg of fresh mass (FM) obtained from E and NE callus were inoculated in Petri dishes containing 20 mL of MS culture medium supplemented with 20 g.L^-1 sucrose, 2 g.L^-1 Phytagel, and varying concentrations of activated charcoal (AC; Sigma-Aldrich; 0.0, 0.75, 1.5 and 3.0 g.L^-1). The pH of the culture medium was adjusted to 5.8 prior to the addition of Phytagel (Sigma-Aldrich) was added. The culture medium was sterilized by autoclaving at 121°C for 15 min. After inoculation, the cultures were maintained in a growth chamber at 25 ± 1°C under dark conditions for the first 7 days. Thereafter, photoperiods of 16 h light (60 μmol.m².s^-1) were used for up to 28 days of culture.
E callus competence was analyzed at 0 (before incubation on maturation treatment), 7, 14, 21 and 28 days of incubation by monitoring the FM (the initial FM was 200 mg FM per colony) and the number of somatic embryos produced.
For both types of callus, three separate Petri dishes, containing three colonies each, were examined per treatment at each sampling time.
Samples of both callus types were collected at 0 and 21days and stored at -20°C for later proteomic analysis.

Heringer A.S., Barroso T., Macedo A.F., Santa-Catarina C., Souza G.H., Floh E.I., de Souza-Filho G.A, & Silveira V. (2015). Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis. PLoS ONE, 10(6), e0127803.

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Seedling Growth under pH Stress

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Five-day-old seedlings grown on 0.5x MS medium with 1% sucrose and 1.2% phytagel (sigma) at pH 5.8 were transferred to 0.5x MS medium of pH 5.8 or pH 8.2 with 1% sucrose and 1.2% phytagel and grow vertically for two more weeks. All the seedlings were placed in a growth chamber at 22 °C under a 16 h–8 h day-night photoperiod. The root length of seedlings was measured in ImageJ.

Guo J., Chai X., Mei Y., Du J., Du H., Shi H., Zhu J.K, & Zhang H. (2022). Acetylproteomics analyses reveal critical features of lysine-ε-acetylation in Arabidopsis and a role of 14-3-3 protein acetylation in alkaline response. Stress Biology, 2(1), 1.

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Callus Production from Estradiol-Inducible cls Plants

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Seedlings of cls-2/cls-2 harboring pER8:CLS plants (here referred to as cls, Katayama and Wada, 2012 (link)) grown on Murashige and Skoog medium (Murashige and Skoog, 1962 (link)) containing 1% (w/v) sucrose and 3% (w/v) phytagel (Sigma–Aldrich, P8169) under continuous light (30 μmol photons m^-2 s^-1) at 23^oC for 1–2 weeks were used for the production of callus materials. When seedlings were large enough to handle, their stems were cut into small pieces, transferred to callus inducing medium [3.1 g L^-1 Gamborg’s B-5 basal salt mixture (Gamborg et al., 1968 (link); Sigma–Aldrich, G5893), 2% [w/v] glucose, 0.5 g L^-1 MES pH 5.7 (KOH), 0.1 g L^-1myo-inositol, 20 mg L^-1 thiamine-HCl, 1 mg L^-1 pyridoxine-HCl, 1 mg L^-1 nicotinic acid, 0.5 mg L^-1 2,4-dichlorophenoxyacetic acid, 0.05 mg L^-1 kinetin, 1 mg L^-1 biotin, 0.5% [w/v] phytagel (Sigma-Aldrich, P8169)], and incubated at 23°C to generate callus. Calli were then transferred to liquid medium and maintained as described previously (Van Gestel and Verbelen, 2002 (link)). Expression of the transgenic copy of CL synthase gene was induced by addition of 17 β-estradiol to fresh MS medium to a final concentration of 4 μM. Cells for mitochondrial preparations were harvested after 6–7 days of sub-culturing.

Petereit J., Katayama K., Lorenz C., Ewert L., Schertl P., Kitsche A., Wada H., Frentzen M., Braun H.P, & Eubel H. (2017). Cardiolipin Supports Respiratory Enzymes in Plants in Different Ways. Frontiers in Plant Science, 8, 72.

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Establishment of Scots Pine Embryogenic Cultures

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One-year-old immature seed cones were collected four times from open-pollinated Scots pine (Pinus sylvestris L.) clone K884 in Punkaharju, Finland (61°48′ N; 29°17′ E) during summer 2004 as described in detail in Vuosku et al. [26 (link)]. Immature seeds were dissected from the developing cones, and the seed coat was removed. Embryogenic cultures were established using immature zygotic embryos with suspensor tissues surrounded by the megagametophyte. The inoculation, induction, and proliferation of the embryogenic cell masses were carried out as described in Sarjala et al. [37 (link)] on a solid basal Douglas-fir cotyledon revised (DCR) medium [38 (link)] modified by Becwar et al. [39 (link)] at room temperature in the dark. In the inoculation and induction stages, DCR medium was gelled by 4 g L ⁻¹ Phytagel™ (Sigma-Aldrich Co., St Louis, MO, USA), whereas in the stage of proliferation, the amount of Phytagel™ was decreased to 2.5 g L ⁻¹.

Muilu-Mäkelä R., Vuosku J., Häggman H, & Sarjala T. (2021). Polyamine Metabolism in Scots Pine Embryogenic Cells under Potassium Deficiency. Cells, 10(5), 1244.

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Germination and Growth of Brachypodium and Arabidopsis

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Brachypodium distachyon (line Bd21) caryopses (grain) were sterilized in 10% NaClO and 0.1% Triton X-100 with gentle shaking for 15 minutes, then washed with sterilized water 5 times. Grain were then placed on wet filter paper in a Petri dish sealed with cling film and incubated for 2 days in the dark at 5°C (B. distachyon) to ensure that all grain germinated at the same time. Grain were placed in Petri dishes containing media as in described previously [25 ] but modified to contain 1% sucrose and 0.5% phytagel (Sigma, UK). Plants were grown vertically in a growth chamber with a 16 hour light period at 23°C for 5 days. Arabidopsis thaliana wild type plants of the ecotype Columbia-0 (Col0) were used. Arabidopsis thaliana seeds were sterilized in 5% NaClO for 5 min and washed with sterilized water for 5 times. Seeds were sown and germinated in Petri dishes containing sterilized media as in Ma et al. [28 (link)] modified to contain 1% sucrose and 0.5% phytagel (Sigma, UK). The plants were grown vertically in the growth chamber with a 16 hour light period at 23°C for 5 days.

Kim C.M, & Dolan L. (2016). ROOT HAIR DEFECTIVE SIX-LIKE Class I Genes Promote Root Hair Development in the Grass Brachypodium distachyon. PLoS Genetics, 12(8), e1006211.

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Evaluating Root Hair and Trichome Phenotypes

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To evaluate root hair phenotypes, plants were germinated and grown on the surface of vertically positioned agar medium containing MS salts, 1% sucrose, and 1% Phytagel^™ (Sigma-Aldrich). Images of the primary root on the agar medium were captured with a CCD camera DFC425 C (LEICA) on stereomicroscope M205 FA (LEICA). The observation and image capture of root surface structures was performed using a microscope Axioplan 2 (Zeiss) equipped with a confocal laser-scanning unit CSU-X1 (Yokogawa) after staining with 5 μg/ml propidium iodide (PI). For root hair lengths, root hairs elongated horizontally to the agar surface in the region 5–7 mm from the root tip were measured on the image with the assistance of ImageJ. Immature root hairs or bulges with length less than 5 μm were omitted. For root hair numbers, both mature and immature root hairs including bulges in the visible side in the region 5–7 mm from the root tip were counted. Trichomes were observed by a variable-pressure scanning electron microscope S-3500 N (Hitachi) equipped with a cooling stage. The 10th–12th leaves were cut from the plants and directly observed invariable-pressure mode (100 Pa) at −20°C. To evaluate trichome and trichome branch numbers, 2-week-old seedlings germinated and grown on the surface of agar medium containing MS salts, 1% sucrose, and 1% Phytagel^™ (Sigma-Aldrich) were used.

Lin Q., Ohashi Y., Kato M., Tsuge T., Gu H., Qu L.J, & Aoyama T. (2015). GLABRA2 Directly Suppresses Basic Helix-Loop-Helix Transcription Factor Genes with Diverse Functions in Root Hair Development. The Plant Cell, 27(10), 2894-2906.

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Evaluating Plant Growth Under Stress

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For growth in soil, plants were grown at 22°C in 75 μE, under long day conditions, with a light regime of 16/8 h. For growth in vitro, seeds were surface-sterilized, sown on bacteriological square petri dishes (120 mm x 120 mm x 17 mm, Greiner Bio-One, 688102) containing1x Murashige and Skoog (MS) medium (Tivan Biotech, MSP01-50LT) with pH adapted to 5.8 using KOH and 0.6% Phytagel (Sigma-Aldrich, P8169). Following cold treatment at 4°C, for 4 d in the dark, plants were grown for 4–7 days in an upright position, at 22°C in 75 μE, under long-day conditions, with a light regime of 16/8 h. For induction of restrictive conditions in vitro, plants were transferred after 4 d in the light to plates maintained in an upright position, with MS medium containing 0.6% Phytagel and one of the following additions: 4.5% sucrose, 100 mM NaCl plus 1% sucrose or the indicated concentrations of Isoxaben (Sigma-Aldrich, 36138) or Fluridon (Sigma-Aldrich, 45511). Plants transferred to MS medium containing 0.6% Phytagel and the corresponding solvent served as control (ambient conditions). The root images were documented using a stereo-microscope (SMZ1270, Nikon) equipped with a camera (NIKON, DS- Ri2).

Allelign Ashagre H., Zaltzman D., Idan-Molakandov A., Romano H., Tzfadia O, & Harpaz-Saad S. (2021). FASCICLIN-LIKE 18 Is a New Player Regulating Root Elongation in Arabidopsis thaliana. Frontiers in Plant Science, 12, 645286.

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Assessing Arabidopsis Growth with Au-NPs

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Arabidopsis thaliana (Columbia ecotype) seeds were grown in gel as described elsewhere. (33) (link)Seeds were surface-sterilized for 5 min in Tween 20, rinsed twice with ethanol, and air-dried sterilely. Ten seeds were sown in square plates (12 × 12 cm 2 ) containing sterile one-half-strength Hoagland's solution solidified with Phytagel (Sigma-Aldrich, United States).
Recipes of these solutions are shown in Table S1 in the SI).
Au-NPs were mixed with the nutrient/Phytagel solution prior to solidification to obtain a concentration of 10.0 mg Au-NPs•L -1 . Control plants without exposure to Au-NPs were also prepared. The plates were sealed with Micropore tape (3M, U.S.A.) to limit water evaporation and allow gas exchange, and incubated vertically for 10 days at a photon flux from the top of 150 μmol•m -2 s -1 and under a light/dark cycle of 16:8 h and 21:19 °C. The germination rate was determined on ten seeds 10 days postsowing, and the length of the roots measured using digital images and ImageJ software.(34)

Avellan A., Schwab F., Masion A., Chaurand P., Borschneck D., Vidal V., Rose J., Santaella C, & Levard C. (2017). Nanoparticle Uptake in Plants: Gold Nanomaterial Localized in Roots of Arabidopsis thaliana by X-ray Computed Nanotomography and Hyperspectral Imaging. Environmental science & technology, 51(15).

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Arabidopsis Oxidative Stress Response Assay

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Arabidopsis thaliana plants were grown on soil (peat: vermiculite = 1:1) under white luminescent light (220-250 µmol m^-2 s^-1) at a 12-hour photoperiod and 22/18 °C. Seedlings were grown for 10 days on 1 x MS basal medium (Sigma) with 0.5 % Phytagel (Sigma). Arabidopsis rcd1-4 mutant (Gabi-Kat line GK-229D11) was used as a background for all complementation lines. The ppk triple mutants were kindly provided by Dr Dmitri Nusinow (Donald Danforth Plant Science Center, St. Louis)^{31 (link)}. Treatments with chemicals methyl viologen (MV, 0.1, 0.5, or 1 μM, as indicated in the figures) and 3-methoxybenzamide (3MB, Sigma, 10 mM) were performed on leaf discs floating on Milli-Q water supplemented with 0.05% Tween 20 (Sigma), overnight at room temperature or at 4 ˚C, accordingly. For 4′,6-diamidino-2-phenylindole (DAPI) staining, the seedlings were vacuum-infiltrated with 0.1 mM DAPI in Milli-Q water supplemented with 0.05% Tween 20.

Vainonen J.P., Gossens R., Krasensky-Wrzaczek J., De Masi R., Danciu I., Puukko T., Battchikova N., Jonak C., Wirthmueller L., Wrzaczek M., Shapiguzov A, & Kangasjärvi J. (2023). Poly(ADP-ribose)-binding protein RCD1 is a plant PARylation reader regulated by Photoregulatory Protein Kinases. Communications Biology, 6, 429.

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Transgenic Arabidopsis thaliana Generation

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Single and double transgenic lines of Arabidopsis thaliana were created through Agrobacterium tumefaciens mediated floral dip transformation (Clough and Bent, 1998 (link)) or through crossing of already selected stable transgenic lines. The proCaMV35S-SFR2mRFP transgenic lines were created in the Columbia ecotype of Arabidopsis thaliana. Other lines used have been published earlier and used for double or triple transgenic plants as detailed in Table 1. All seeds were stratified for 2 days at 4°C. For investigating soil-grown plants Sunshine mix LA4 (Sun Gro Horticulture, United States) soil in sealed Magenta boxes was used. Alternatively, if specified, plants were grown in plastic petri dishes containing Murashige and Skoog basal medium (Murashige and Skoog, 1962 (link); Sigma M404) with B5 vitamins, 0.75% sucrose and 3 g/L phytagel (Sigma-Aldrich) (pH adjusted to 5.8 before autoclaving). Plants were grown for 7–14 days under a long-day light regime (16-h-light–8-h- dark) under light intensity of 120 ± 20 μmol m^–2 s^–1 light and an ambient temperature of 21 ± 2°C.

Mathur J., Kroeker O.F., Lobbezoo M, & Mathur N. (2022). The ER Is a Common Mediator for the Behavior and Interactions of Other Organelles. Frontiers in Plant Science, 13, 846970.

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