![]() ![]() We advise always using more than one sample when trying to induce protonemata or isolate protoplasts. While growth conditions for Sphagnum gametophores have been established for many species, induction of protonemata can be difficult. Adjust osmolarity via mannitol concentration ± 5 mOsm. For this, a freezing point depression osmometer was used. Osmolarity Osmolarity should be checked using an osmometer with several measurements. Should be sterilized by filtration with 0.22 µm bottle top filter to assure stability of pH and osmolarity. Regeneration medium (RM): Knop ME medium with 50 g/L glucose monohydrate, 20 g/L mannitol, pH 5.5, 440 mOsm/L. Protoplast isolation medium (PIM): 80 g/L mannitol, pH 5.7, 450 mOsm/L. Knop ME pH 4.5: Knop ME medium, 0.1% MES, pH 4.5, cultivation at 22 ☌, 16 h light/8 h dark, light intensity 70 ± 5 µmol/s. Protonema medium (PM): Knop ME pH 5.4 with 30 mM ammonium tartrate. Knop ME pH 5.4: standard moss medium, described in Beike et al. Suspension culture for protonema induction and pre-culture prior to protoplast isolation were performed in 50 mL and 200 mL of media. ( 2021a).Īll cultivation techniques used for Sphagnum gametophores are described in Beike et al. Protocols Media, moss lines and cultivationĪxenic Sphagnum strains are available from the International Moss Stock Center ( and published in Heck et al. Moreover, we provide protocols for protoplast isolation from protonemata, the first protocol for transformation of Sphagnum cells and protoplast regeneration without feeder cells. Here, we report on an efficient method for protonema induction, which works in different Sphagnum species, as well as long-term cultivation, preventing differentiation of gametophores, and allowing growth of homogenous, thalloid protonemata in suspension. So far, regeneration of Sphagnum protoplasts has been achieved only with a chlorophyll-free Solanum hybrid clone as feeder (Batra et al. This is laborious and contrary to other mosses, where protoplasts are isolated from protonemata as an “easier to handle” material, as protonemata are grown in suspension culture in different volumes (Hohe and Reski 2002 Schween et al. So far, protoplasts were isolated from gametophore apices of S. squarrosum had to be sub-cultivated regularly to prevent gametophore formation. Upon wounding, gametophores will eventually form thalloid protonemata, a method described in Zhao et al. 2021a, b), juvenile protonema tissue, from which the gametophores arise, is naturally formed only after spore germination (Clymo and Hayward 1982 Beike et al. While Sphagnum gametophores are easy to propagate (Beike et al. 2021b), the toolbox for working with Sphagnum mosses under axenic conditions is still small. 2021a) and cultivation techniques available (Batra et al. While there are a number of axenic Sphagnum laboratory strains (Heck et al. As climate-critical plants, Sphagnum mosses have gained interest in recent years (Beike et al. Amphibians.Peat mosses of the genus Sphagnum comprise most of the biomass in northern peatlands, storing over 500 Gt of carbon, while covering only 3% of the earth’s land mass (Gorham 1991 Clymo et al. Use: Top dressing or soil mix for carnivorous plants. Photographs are representative of the species, and not the specific plant shipped. A portion is 1 measuring cup full, enough to cover a 4" pot. Keep very moist and water with rain/distilled water. Though Red Sphagnum Moss has no roots, it benefits from planting in a loose CP soil mix. It is a shorter, slower growing Sphagnum, and does well with all but the smaller CPs. It spreads nicely, and can be divided into smaller plugs to plant a larger area. It prefers the open, heathy areas of the bog, often in areas with limestone bedrock. Unlike more common forms of lawn or woodland moss, Sphagnum Moss is larger, and requires very damp growing conditions. Like other Sphagnum Mosses, Red Sphagnum Moss is a primitive bryophyte like ferns and clubmosses. The intensity of color seems to favor bright, cooler conditions. The short stems have sweeping, outward-curving branches that resemble small tresses. It tends to form tight, carpet-like mounds or clumps resembling red cauliflower. It grows in the same conditions as the typical form, tolerating less acidity and more light. These clones are from maroon (dark red) selections. Color can vary from salmon pink to maroon, almost black. Red Sphagnum Moss, Sphagnum sp., or the Small Red Peat Moss, is a beautiful and unusual dark red form of Sphagnum.
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