The rapid propagation technology using shoot tip tissue culture is an effective method to eliminate or reduce viral infections in lotus roots, restore the desirable characteristics of specific varieties, enhance the propagation rate, and speed up the dissemination of high-quality strains. In recent years, Xiangshui Shallow Water Science and Technology Demonstration Park has successfully implemented this technique, gradually achieving industrial-scale production of shallow water seedlings.
First, initial culture begins in mid-April when lotus roots start sprouting. Fresh seedlings are selected for explant culture. The stem tips (including the apical bud) are carefully taken, disinfected, and the outer sheaths are removed using forceps. When the stem tips reach a diameter of about 2 mm, they are inoculated into a differentiation medium. After approximately 30 days of cultivation, multi-leaf seedlings can be obtained through differentiation.
Second, proliferation and rooting culture follow.
1. Subculture and propagation: The primary culture seedlings are cut under sterile conditions and divided into three types of explants—those with one spread-out leaf, one unexpanded leaf, and one shoot. These are then placed individually into a proliferation medium. After about 30 days, the cells are harvested and transferred to a basal culture for further propagation. It's recommended that the proliferation phase last around 30 days. Under optimal conditions, four generations can be completed within 120 days, achieving a theoretical propagation coefficient of over 2000 times.
2. Rooting culture: Once a sufficient number of subcultured seedlings are ready, they are moved to a basal culture and then transferred to a rooting medium under sterile conditions. After 30 to 40 days, well-developed root systems form, resulting in complete plantlets suitable for vegetative propagation.
Third, acclimatization and transplanting.
1. Acclimatization: Rooted seedlings are moved from the lab environment to the field together with their bottles, kept at around 25°C. They are shaded, misted, and allowed to gradually adapt by opening the bottles for a week. This helps them adjust to outdoor conditions.
2. Transplanting: Carefully remove the seedlings from the bottles, rinse them with water, and sterilize the soil in a shallow layer of 3–5 cm. Plant them with a row spacing of 1 meter and a plant spacing of about 0.5 meters. Maintain a temperature around 25°C with appropriate shading. After about 15 days, the seedlings will grow new leaves. With proper field management, they will continue to develop. Under open-field conditions, the seedlings need a growth period of at least 100 days with temperatures above 15°C before they can produce larger rhizomes for planting the following year.
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