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#plant culture techniques

3 public questions tagged with this topic.

Plant tissue culture 1

Most widely used plant tissue culture medium is:

Murashige and Skoog medium proposed in 1962 for tobacco pith culture remains gold standard and most widely used basal medium for plant tissue culture due to broad applicability and high salt formulation supporting rapid growth across herbaceous and many woody species. Its composition features high concentrations of macronutrients ammonium nitrate 1650 mg per L and potassium nitrate 1900 mg per L providing dual nitrogen source favoring both amino acid synthesis and pH buffering, plus calcium chloride, magnesium sulfate, potassium dihydrogen phosphate; chelated iron as Fe-EDTA, micronutrients ma

Ref: Murashige & Skoog Physiol Plant 1962; NCBI NBK26873 MS medium composition.

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Plant tissue culture 1

Ideal aseptic condition means:

Ideal aseptic condition means complete absence of viable bacteria, fungi, yeast, algal spores and viruses from all components including culture vessel, nutrient medium, explant surface and environmental air, producing axenic system. Tissue culture media enriched with 30 grams per liter sucrose, vitamins, amino acids and growth regulators represent perfect substrate for microbial proliferation; even few contaminants rapidly multiply outcompeting slow-growing plant cells for nutrients, alter pH through fermentation, release phytotoxic metabolites like hydrogen sulfide, enzymes degrading plant ce

Ref: CDC Guidelines for Aseptic Technique; Bhojwani Plant Tissue Culture manual, Chap. 2.

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Plant tissue culture 1

Surface sterilization of explants commonly uses:

Surface sterilization removes epiphytic and surface-borne microorganisms from explants without killing plant tissues, essential prerequisite for establishing contaminant-free cultures. Most effective protocol uses sequential treatment with 70 percent ethanol for 30-60 seconds followed by sodium hypochlorite solution 0.5-2 percent available chlorine for 10-20 minutes containing surfactant Tween-20 0.1 percent to improve wetting. Ethanol acts as degreasing agent dissolving epicuticular waxes, disrupting microbial lipid membranes and denaturing proteins facilitating penetration of subsequent ster

Ref: Bhojwani & Dantu Plant Tissue Culture, Chap. 3 sterilization; NCBI StatPearls Aseptic Technique.

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