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| Product Name | Plant Tissue Culture Antimicrobial Preservative |
| Catalog No. | RESK-0599 |
| Description | Microbial contamination is the greatest concern throughout plant tissue culture, from callus induction through root and shoot regeneration, whether in solid, plate or liquid (static, suspension or floating) culture systems. This preservative is a broad-spectrum antimicrobial agent that kills bacterial and fungal cells, prevents the germination of fungal spores and, at higher concentrations, eliminates endogenous contamination within explants. Its active ingredients penetrate fungal and bacterial cell walls, inhibit key enzymes of central metabolic cycles such as the citric acid cycle and the electron transport chain, and block the transport of monosaccharides and amino acids from the medium into microbial cells. It effectively suppresses airborne, water-borne, human-borne and endogenous microbial contamination in plant cell and tissue culture without affecting in vitro seed germination, callus proliferation or callus regeneration. |
| Intended Use | Broad-spectrum antibacterial and antifungal preservative for the prevention and elimination of microbial contamination in plant tissue and cell culture, including treatment of endogenously contaminated explants and removal of Agrobacterium after co-cultivation. For research use only. |
| Principle / Technology | The active ingredients penetrate microbial cell walls and inhibit key enzymes in central metabolic cycles (citric acid cycle and electron transport chain) while blocking uptake of monosaccharides and amino acids from the culture medium. Because it targets multiple enzymes, the emergence of resistant mutants is effectively impossible. |
| Sample Type | Plant tissue culture media and explants, including callus, seeds, tubers, bulbs, corms and transformed leaf discs. |
| Performance Range / Specifications | Advantages over conventional antibiotics: 1) Broad-spectrum, high-efficiency antibacterial/antifungal activity. 2) Less expensive than commonly used antibiotics; can be used routinely as a standard component of plant tissue culture media. 3) Because it acts by inhibiting multiple enzymes, the generation of resistant mutants is impossible. 4) Heat-stable and can be sterilized together with the culture medium. |
| Specificity | Active against bacterial and fungal contaminants of plant tissue culture; does not affect in vitro seed germination, callus proliferation or callus regeneration at recommended doses. |
| Reaction Conditions / Protocol | The following procedures are general and may be modified for specific species. 1. Media containing the preservative can be dispensed outside the clean bench (no aseptic operation required); cap after the agar solidifies. If a pump is used, flush the tubing with autoclaved hot water before and after dispensing. 2. If stored in sterile containers and uncontaminated, preservative-containing heat-sensitive or heat-stable liquid media require neither membrane filtration nor autoclaving; for enriched media containing 200 mg/L or more amino acids or protein, filter after adding the preservative. 3. Instruments (forceps, scalpels) need not be flamed; dip periodically in 70% alcohol. Work can be performed on a clean bench outside the laminar flow hood for up to one hour. 4. The solution is acidic (pH 3.8) and should be stored at 4 °C. At low inoculum density the recommended working dose is 0.05-0.2% (v/v), added before or after medium sterilization, to prevent airborne and endogenous contamination. Higher doses are required to treat endogenous contamination or to obtain Agrobacterium-free plant material. 5. For seeds heavily contaminated on the epidermis with bacteria or fungal spores, surface-sterilize with bleach by the conventional method before in vitro germination; seeds can then be rinsed under tap water in a non-sterile container and air-dried in the clean bench before placement on preservative-containing germination medium. Instruments contacting active bacteria, fungi or suspected contaminants should be sterilized by autoclaving or electric heating. 6. General dose levels: except for endogenous contamination, 0.05-0.2% is recommended; for callus proliferation, organogenesis and embryogenesis, 0.05-0.075% is recommended. 7. Endogenous contamination: a) For explants of 1 cm or shorter, add 4-5% preservative to full-strength MS basal salts (do NOT add Tween 20 or pH adjusters), agitate for 4-12 h, then insert directly without rinsing into preservative-containing medium; transfer herbaceous plants to germination medium with 0.05-0.1% and woody plants to 0.2%. b) For tubers, bulbs and corms: agitate the whole organ in bleach, rinse with water (non-sterile conditions acceptable), cut into small pieces, agitate in full-strength basal salts with 4-5% preservative for 12-24 h (no pH adjusters or Tween 20), and insert without rinsing into medium containing 0.1-0.2% preservative. 8. If the above gives unsatisfactory results (especially thick or very dense explants and seeds): a) agitate explants in water (1 h for soft tissue, 2 h for hard tissue); b) agitate in full-strength MS basal salts containing 50% preservative (no pH adjusters or Tween 20) for 5-10 min; c) insert directly into medium without rinsing. For fungal contamination, additional preservative may be added; for bacterial or mixed contamination, 0.05-0.2% in the medium is necessary for the first month. Do not discard highly oxidized explants; about 50% recover within 4-6 weeks. 9. Rescue of plant material contaminated during culture (only for material visibly contaminated for no more than one week): a) scrub under a gentle stream of tap water with a toothbrush; agitate in 50% preservative (diluted with sterile water) for 5-15 min. For bacterial or mixed contamination, use a low-pH (2.8-3.2) solution: 100% preservative mixed 1:1 with 0.6 g/L citric acid in sterile water. b) Insert without rinsing into medium containing 0.05-0.2% preservative and culture for at least one month, with low light for the first 10 days. For contaminants deep within the explant, soak, section along the explant and agitate in 50% preservative for 5-15 min to ensure full surface contact. 10. Agrobacterium removal: after co-cultivation, rinse leaf discs with water, dip the whole disc in 100% preservative in full-strength basal salts for about 2 min, blot dry on sterile paper towels and place on medium with conventional antibiotics; after three weeks transfer to medium containing only 0.05-0.075% preservative. |
| Components / Formulation | Proprietary broad-spectrum antimicrobial formulation; acidic aqueous solution, pH 3.8. |
| Storage Conditions | Store at 4 °C. |
| Shelf Life | 2 years |
| Package Specifications | 25 mL, 100 mL |
| Product Form | Liquid (acidic aqueous solution) |
| Quality Control | The product itself is non-sterile; filter sterilization is recommended before use, and it can also be heat-sterilized. |
| pH Range / Optimal pH | pH 3.8 (the preservative solution is acidic); a working pH of 2.8-3.2 is used for rescue treatment of bacterial or mixed contamination. |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 2 years when stored at 4 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Heat-stable; can be autoclaved together with plant tissue culture media (e.g., MS basal salts). Do not combine with Tween 20 or pH adjusters during explant pretreatment. |
| Recommended Buffer System | Full-strength MS (Murashige and Skoog) basal salts for explant pretreatment solutions. |
| Application Notes / Precautions | Recommended working dose is 0.05-0.2% (v/v), added before or after medium sterilization; 0.05-0.075% for callus proliferation, organogenesis and embryogenesis. Higher doses (4-5%, 50% or 100% in pretreatment solutions) are used for endogenous contamination, rescue and Agrobacterium removal. Never add Tween 20 or pH adjusters to pretreatment solutions. Do not discard highly oxidized explants; approximately 50% recover within 4-6 weeks. The product is non-sterile; filter sterilization or heat sterilization is recommended. |
For research use only, not for clinical use.
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