© Name: Hema © Roll No. : 230013 © Subject : Application of Plant tissue and culture © Topic : Micropropogation of Aloe Vera © Submitted to: Priyanka Mishra MamIntroduction ~ Micropropagation is a modern biotechnological method of rapidly multiplying plant material using tissue culture techniques. It involves growing plant cells, tissues, or organs ina controlled sterile environment on a nutrient culture medium. This technique is especially useful for plants like Aloe vera, which are economically and medicinally important. Through micropropagation, thousands of identical and disease-free Aloe vera plants can be produced in a relatively short time, compared to traditional propagation methods. 2. Botanical Description of Aloe vera off Aloe vera (Aloe barbadensis Miller) is a stemless or short-stemmed succulent plant that grows up to 60-100 cm tall. Its leaves are thick and fleshy, green to grey-green, with serrated edges. Inside the leavesis a clear, jelly-like gel widely known for its healing properties. Family: Asphodelaceae Habitat: Prefers dry, arid climates; drought-resistant Reproduction: Naturally by suckers and offshoots, but micropropagation provides a faster alternative. 3. Importance of Aloe vera Aloe verais globally recognized for its medicinal, cosmetic, and nutritional value: Medicinal use: Treats burns, wounds, and skin irritations; sup ports digestion and immunity Cosmetic use: Common ingredient in skincare products like lotions, gels, and creams Nutritional use: Juice consumed as a detoxifying drink Industrial use: Used in pharmaceuticals, food supplements, and personal care productsWhat is Micropropagation? Micropropagation is a technique that allows the propagation of plants under controlled laboratory conditions using small pieces of plant tissue (explants). It ensures rapid multiplication of genetically identical, disease-free plantlets. The process uses sterilized conditions, growth hormones, and nutrient-rich culture media (such as MS medium) Key Features: Can multiply plants in a limited space. Allows propagation of plants that are hard to grow through seeds or cuttings. Used widely in agriculture, horticulture, and conservation. Stages of Micropropagation Stage I: Initiation Selection of a healthy explant (shoot tip, leaf base, or sucker)Surface sterilization using chemicals (e.g., ethanol, sodium hypochlorite) inoculation onto sterile culture medium (Murashige and Skoog - MS mediurn). Objective: Establish contamination-free growth Stage II: Multiplication Explants are transferred to media containing cytokinins like BAP (Benzylaminopurine) to induce shoot proliferationMultiple shoots. grow from one explantThis stage is repeated to increase shoot numbers. Stage Ill: Rooting Individual shoots are placed on rooting medium with auxins like IBA (Indole-3-butyric acid) or NAARoots form, producing complete plantlets.Plantlets are ready for transfer to natural conditions. Stage IV: Acclimatization Gradual transfer of plantlets to external environment.initially grown in a greenhouse under controlled humidity and temperature.After hardening, they are transplanted to soil in fields or pots. Micropropagation in Aloe vera @ In Aloe vera, conventional propagation through offshoots is slow and often yields limited plants. Micropropagation overcomes this by allowing the mass production of uniform and vigorousplants.Common Explants: Shoot tips, leaf segments, axillary buds. Culture Medium: MS medium with BAP (for shoot proliferation) and IBA (for root induction) Results: High multiplication rate, uniform morphology, and stable biochemical content. Used in pharmaceutical industries to meet the high demand for Aloe vera. Detailed Protocol for Aloe vera Micropropagation A. Selection and Sterilization of Explants Explants: Shoot tips or young leaf bases Sterilization steps: 4. Wash under tap water for 10-15 mins 2. 5% detergent (Tween-20) for 5 mins 3. 70% ethanol for 30 seconds: 4.0.1% HgCl for 5 mins 5. Rinse 3-4 times with sterile distilled water B. Culture Medium Preparation Medium: Murashige and Skoog (MS) medium Additives: 3% sucrose 0.8% agar BAP (2.0 mg/L) for shoot inductian NAA (0.5-1.0 mg/L) for rooting pH; Adjust to 5.8 before autoclaving C. Culture Conditions Temperature: 24—26°C Photoperiod: 16-hour light / 8-hour dark Light intensity: 3000-4000 lux Humidity: 50-60%Advantages of Micropropagation + 1. High multiplication rate: Thousands of plants from one explant. 2, Disease-free plants: Sterile lab conditions eliminate pathogens. 3. Uniformity: Genetically identical plants with consistent traits. 4, Faster propagation: Especially for slow-growing or rare plants. 5, Space-efficient: Large numbers grown in small lab spaces. 6, Year-round production: Independent of seasons or climate. Challenges in Micropropagation A 1. Costly setup: Requires equipment, lab facilities, and trained personnel. 2.Risk of contamination: Fungal or bacterial infections can ruin cultures. 3. Somaclonal variation: Genetic changes may occur if hormone levels are not optimized. 4, Labor-intensive: Requires meticulous handling. 5. Acclimatization loss: Improper transition to field conditions can lead to plant death. Applications of Micropropagation 4 1.Commercial farming: Large-scale production of Aloe vera for sale. 2.Pharmaceutical industry: Reliable source of medicinal plants. 3.Cosmetics: Consistent supply of gel-rich plants for product manufacturing. 4.Conservation biology: Preserving endangered or rare plant species. ‘5.Res earch and breeding: Producing genetically identical plants for experiments.* Conclusion Micropropagation offers an efficient and reliable method for the large-scale cultivation of Aloe vera, meeting the demands of pharmaceutical, herbal, and cosmetic industries. With proper protocol and care, this technique can produce high-quality, disease-free plants rapidly and sustainably. . References 1. Murashige, T., & Skoog, F. (1962). Physiologia Plantarum, 15(3), 473-497. 2. Kumar, S., & Reddy, M.P. (2011). Plant Tissue Culture and Biotechnology, 21(2), 135-141. 3. Rout, G.R., & Das, P. (2002). Plant Cell, Tissue and Organ Culture, 71, 277-285. 4. George, E.F,, et al. (2008). Plant Propagation by Tissue Culture. Springer. 5. Sharma, A., & Chandel, K.P.S. (1992). Plant Cell Rep orts, 11(3), 133-136.