The ph of nutrient medium for plant tissue culture is in the range of

Autoclaving is a standard procedure for sterilizing nutrient media for plant tissue cultures. Most tissue cultures are grown at pH 5.2 to 5.8 with pH adjustments being made prior to autoclaving. This paper reports that there are significant differences between initial pH levels and pH levels following autoclaving, particularly in the pH range of 5.7 to 8.5. This effect is noted with and without agar. In addition, we report that with time the pH of the medium drifts into the acid range. When Cucumis callus was added to the medium, the pH was changed significantly within 48 hours. The amount and direction (increase or decrease of pH) was significantly correlated with the original pH. This suggests that researchers should be wary of the true pH situation in their medium. In addition, in publications authors should specify whether their medium pH value was determined before or after autoclaving. • Anderson WC (1975) Proc Int Plant Prop Soc 25:129 • Behagel HA (1971) The pH and sterilization. In: Van Bragt J, Mossel DAA, Pierik RLM, Veldstra H (eds) Effects of sterilization on components in nutrient media. Veenman H and Zonen N.V. Netherlands, pp 117–120 • Bridson EY (1978) Diets, culture media, and food supplements. In: Rechcigl M Jr (ed) Handbook series in nutrition and food, vol III, section G, Chemical Rubber Company Press, Boca Raton, Florida, pp 148–149 • Cousson A, Tran Thanh Van K (1981) Physiol Plant 51:77–84 • Dougall DK (1980) Nutrition and metabolism. In: Staba EJ (...

Murashige and Skoog Medium Modified MS medium lacking ammonium, nitrate and pyridoxine hydrochloride, and supplemented by indoleacetic acid (0.5 p.p.m.), 2,4-dichlorophenoxyacetic acid (1 p.p.m.) and benzyladenine (0.5 p.p.m.), was the best medium. From: Genetic Improvement of Vegetable Crops, 1993 Related terms: • Kinetin • Indole-3-Butyric Acid • 1-Naphthaleneacetic Acid • Tissues • Buds • Callus • Explant 1. Liquid MS medium (Murashige and Skoog (MS) basal medium with vitamins 0.44% w/v (as described by Murashige and Skoog ( Murashige & Skoog, 1962)), Sucrose 2% w/v, myo-Inositol 0.01% w/v at pH 5.8). a. Weigh 4.4g of MS powder, 20g of sucrose, 100mg of myo-Inositol and add 900mL of deionized water. Stir the solution at room temperature until all components are dissolved, then adjust the pH to 5.8 with KOH. Make up to 1L with deionized water and autoclave. Store at room temperature. 2. Solid MS medium (MS basal medium with vitamins 0.44% w/v, Sucrose 2% w/v, myo-Inositol 0.01% w/v, agar 0.7% w/v, pH 5.8). a. The procedure is the same as the previous one (see Section 2.3, #1), but after adjusting pH, 7g/L agar is added and then the medium is sterilized. Store at room temperature. 3. MI (embryo induction medium (Solid MS media plates with 1mg/L 2,4-dichlorophenoxyacetic acid (2,4-D)). a. Let the solid MS medium (see Section 2.3, #2) cool after sterilization and then add 0.5mL of 2,4-D (1mg/mL) to 500mL of warm medium. Mix the medium very gently to avoid generating bubbles...

This review presents an overview of the culture media and practices used in plant tissue culture and developmental biology. The compositions of the most commonly used basal media, especially Murashige and Skoog (MS) and modified MS (MMS), Gamborg’s B5 medium and B5 modifications, Woody Plant Medium (WPM), and Driver and Kuniyuki Woody plant medium (DKW) are discussed, along with typical basal medium manipulations to elicit and support various developmental responses. The most commonly used plant growth regulators and their applications to promote various developmental responses are examined, along with a presentation of the classical phytohormone developmental models for organogenesis and somatic embryogenesis. Elaborated developmental models for both organogenesis and somatic embryogenesis, with emphasis on discrete developmental steps, occasional need for multiple manipulations in culture to achieve a single developmental step, and identification of responsive tissue types in mixed cultures are explored. It is hoped that the information presented here will lead to a deeper understanding of basic tissue culture responses and will assist the reader in the decision-making process by identifying appropriate media and culture conditions for a particular species or application, or by providing a suitable starting point, should further customization be required. • Armstrong CL, Green CE (1985) Establishment and maintenance of friable, embryogenic maize callus and the involvemen...

Abstract Medium pH is generally adjusted to 5.8 to 6.0 for plant tissue culture. Our research indicated that pH generally falls between 5.5 and 7.5 in an ordinarily made medium which can be directly used for apple tissue culture without adjusting pH. Repeated adjustment of pH by adding NaOH and HCl leads to the increase in Na + and Cl − concentration and decrease in Mg 2+ and Ca 2+ concentration in the medium due to precipitation. To determine the pros, cons, and necessity of pH levels while making medium in plant tissue culture, subculture proliferation, adventitious root induction, and organ regeneration, the apple cultivars Fuji, Golden Delicious, Jonagold, and Gala were used and hardness of the medium and the ion content of Na +, Cl −, Mg 2+, and Ca 2+ in the medium under different pH were measured. In the lower pH range of 5.0–5.5, plantlets could be subcultured and grew normally; however, the medium did not solidify or solidified poorly resulting in problems associated with handling. No significant difference was found among the treatments when pH ranged 6.0–8.0 in terms of proliferation, adventitious root induction, and adventitious bud regeneration from leaves, except a slight decrease in shoot number proliferation in ‘Jonagold’ and in adventitious bud regeneration from leaves in ‘Fuji’ and ‘Golden Delicious’ at pH above 7.5. The hardness of the medium increased with the increased pH. The superfluous Cl − and Na + generated during the process of overadjusting pH to...

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