Sharabati Medium: A Comprehensive Guide to Its Use and Formulation in Plant Tissue Culture
Tissue culture media play a crucial role in the successful propagation, genetic modification, and research of plants at the cellular and tissue level. Every medium is carefully designed with specific components aimed to support the growth of cultured cells, tissues, or organs. One such specialized medium used in plant tissue culture is the "Sharabati Medium." Though not as commonly discussed in mainstream tissue culture resources, this medium holds unique applications in plant science. In this blog post, we will explore what Sharabati Medium is used for and provide its detailed formulation.
What is Sharabati Medium?
Sharabati Medium is a plant tissue culture medium specifically formulated for use in the micropropagation of certain plant species. It has been designed to facilitate the in vitro growth, differentiation, and development of difficult-to-culture plants, especially those requiring a finely tuned balance of nutrients and growth regulators. This medium is primarily used for the culture of recalcitrant plants or those that are hard to regenerate through more conventional tissue culture methods.
This medium’s unique composition provides a controlled environment that supports the following processes in plant tissue cultures:
- Callus Induction: Encouraging the growth of callus tissue from explants.
- Somatic Embryogenesis: Promoting the development of somatic cells into embryoids that eventually grow into full plants.
- Rooting and Shoot Formation: Facilitating cell differentiation into root and shoot meristems for whole plant regeneration.
While this medium is understandably tailored to specific plant species, it has found importance in research and applied plant biotechnology sectors focusing on conservation, plant breeding, and plant propagation.
Key Uses of Sharabati Medium
Some major applications of Sharabati Medium include:
Micropropagation of Rare and Endangered Plant Species: Often plants that are difficult to propagate naturally or via traditional micropropagation methods are cultured on Sharabati Medium.
Genetic Conservation & Revitalization of Germplasm: Some plants that are at risk of extinction are cultured in vitro using Sharabati Medium to preserve their genetic material.
Research in Plant Stress Tolerance: Since this medium is adaptable to different environmental conditions, it’s used to study plant stress responses, including salt and drought resistance.
- Plant Breeding Programs: Sharabati Medium helps in improving the efficiency of breeding programs by providing consistent plant regeneration from tissue cultures, allowing faster selections for desired traits.
The Formulation of Sharabati Medium (per 1 Litre)
The precise formulation of Sharabati Medium may vary based on plant species and research purposes. However, the core components listed below provide the essential mix of nutrients, carbohydrates, vitamins, and growth hormones to sustain plant tissue:
| Component | Concentration |
|---|---|
| Macronutrients: | |
| NH₄NO₃ (Ammonium Nitrate) | 1650 mg |
| KNO₃ (Potassium Nitrate) | 1900 mg |
| CaCl₂·2H₂O (Calcium Chloride) | 440 mg |
| MgSO₄·7H₂O (Magnesium Sulfate) | 370 mg |
| KH₂PO₄ (Potassium Phosphate) | 170 mg |
| Micronutrients: | |
| MnSO₄·H₂O (Manganese Sulfate) | 22.3 mg |
| ZnSO₄·7H₂O (Zinc Sulfate) | 8.6 mg |
| H₃BO₃ (Boric Acid) | 6.2 mg |
| KI (Potassium Iodide) | 0.83 mg |
| Na₂MoO₄·2H₂O (Sodium Molybdate) | 0.25 mg |
| CuSO₄·5H₂O (Copper Sulfate) | 0.025 mg |
| CoCl₂·6H₂O (Cobalt Chloride) | 0.025 mg |
| Iron Source: | |
| FeSO₄·7H₂O (Ferrous Sulfate) | 27.8 mg |
| Na₂-EDTA (Disodium EDTA) | 37.3 mg |
| Vitamins: | |
| Thiamine-HCl (Vitamin B1) | 0.4 mg |
| Pyridoxine-HCl (Vitamin B6) | 0.5 mg |
| Nicotinic Acid (Niacin) | 0.5 mg |
| Plant Growth Regulators: | |
| 6-Benzylaminopurine (BAP) [Cytokinin] | 1.0 mg |
| Indole-3-Acetic Acid (IAA) [Auxin] | 0.1 mg |
| Carbohydrate Source: | |
| Sucrose | 30,000 mg (30 g) |
| Gelling Agent: | (optional, for solid media) |
| Agar or Phytagel | 8,000 mg (8 g) |
pH Adjustment: The pH of the medium should be adjusted to around 5.6 to 5.8 using KOH or HCl before autoclaving at 121°C for 20 minutes.
Note that growth hormones such as BAP (a cytokinin) and IAA (an auxin) need to be optimized based on the specific plant species and desired tissue response (callus formation, root induction, shoot regeneration, etc.). The presence of these growth regulators is a critical component in determining how cells will differentiate during the tissue culture process.
Additional Notes on Ingredient Roles
- Macronutrients: Provide essential nitrogen, potassium, calcium, sulfur, and phosphorus to cells, encouraging growth and metabolic function.
- Micronutrients: Trace elements that assist in enzymatic processes and overall cellular health.
- Iron Source: Both ferrous sulfate and EDTA are used in combination to provide plants with a stable source of iron, preventing chlorosis (yellowing of leaves).
- Sucrose: Serves as the primary carbohydrate, supplying energy for cellular metabolism.
- Growth Regulators (BAP, IAA): These determine plant morphogenesis in vitro, inducing the right balance of growth based on the desired tissue development.
Conclusion
Sharabati Medium may not be as widely discussed as media like the Murashige and Skoog (MS) medium, but it fills a niche role in plant tissue culture, particularly for species that need a more specialized nutrient balance. Whether you’re working on rare species conservation or looking for ways to unlock growth in plants that have resisted previous attempts in vitro, Sharabati Medium offers a proven solution.
By understanding the formulation and functions of its individual components, you can effectively tailor the medium for a wide range of plant propagation uses, from breeding programs to advanced conservation efforts.
If you’re in the world of plant tissue culture and haven’t yet experimented with Sharabati Medium, it’s worth considering for your next in vitro endeavor!
Got any questions about Sharabati Medium or tissue culture in general? Drop them in the comments below, and let’s keep the conversation growing!