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Aloe Tissue Culture Rapid Propagation
Aloe vera, a perennial evergreen succulent herb belonging to the genus Aloe, has gained widespread attention in recent years due to extensive research in its chemistry and pharmacology. This has sparked a global trend in aloe-based health care products. In China, the cultivation of aloe has started to grow rapidly, but the plant is unable to self-pollinate, making seed propagation difficult and inefficient. Current methods mainly rely on vegetative reproduction through ramets and tillers, which limits large-scale and rapid propagation. As a result, aloe seedlings remain expensive. To address this issue, this article introduces an efficient technique for the rapid propagation of aloe seedlings using tissue culture of stem tips. This method allows for the mass production of millions of seedlings in a short time, offering a low-cost and highly efficient solution.
**Materials and Media**
Healthy aloe seedlings with a height of 10-15 cm are used as explants. The callus induction medium consists of MS basal medium supplemented with 1–6 mg/L BA and 0.1–0.5 mg/L NAA. For differentiation, the medium contains 2–4 mg/L BA and 0.1–0.5 mg/L NAA. Rooting medium includes MS with 0.1–0.5 mg/L IBA and 2–5 mg/L PP33.
**Breeding Method**
(1) **Callus Culture**: Aloe seedlings are washed thoroughly in clean water, and the stem tips are carefully selected after removing leaves and most of the stem. The tips are surface-sterilized with 75% ethanol for 30–60 seconds, followed by 5–10 minutes in mercury solution, and then rinsed multiple times with sterile water. The shoot tip, including the growth point, is excised and inoculated onto the callus induction medium. Cultures are maintained under 10–12 hours of light per day at 1000–2000 Lx, with a temperature of 25 ± 2°C. After 15–25 days, callus formation is observed, indicating the initiation of cell division.
(2) **Subculture**: Once callus forms, it is transferred to fresh medium. The callus is cut into small pieces and re-inoculated. This process can be repeated to generate more material for further culture.
(3) **Differentiation Culture**: Callus is transferred to the differentiation medium, where it develops buds and small leaves over 30–50 days. When the seedlings reach a suitable size, they are moved to rooting medium.
(4) **Rooting Culture**: Seedlings with 3 cm long leaves are placed in rooting medium. Roots typically appear within 15 days.
(5) **Hardening Off**: Once roots develop, seedlings are removed from the test tube, washed in clean water, and kept in a controlled environment (15–25°C, 60–80% humidity) for 12–24 hours before transplanting.
(6) **Transplanting**: Seedlings are transplanted into vermiculite-based nurseries. They are misted daily with a nutrient solution and fresh water. After 15–20 days, they are ready for field planting.
**Important Notes**
1. All reagents must be accurately measured. Sucrose can be replaced with white sugar to reduce costs. Sterilization is done at 120°C, 1.5 kg/cm² for 30 minutes. The culture room should be fumigated with formaldehyde and sterilized with UV before each use.
2. If contamination occurs, affected cultures should be discarded. Only healthy parts can be re-cultured if contamination is localized.
3. After hardening, seedlings can be soaked in a rooting powder solution before transplanting. During domestication, maintain proper humidity, temperature, and light. Common pests like spider mites and aphids can be controlled with 40% omethoate EC diluted 1200 times. Fungal diseases such as black spot can be treated with 50% carbendazim WP diluted 1000 times.
This method offers a reliable and scalable approach to aloe propagation, supporting sustainable and cost-effective cultivation.