Spring Orchid Tissue Culture for Rapid Propagation of Dendrobium Nobile

⑴ We established a small family organization culture room and manufactured our own equipment. As is known, this work must be carried out under sterile conditions. To achieve this, we developed well-sealed glass inoculation boxes, purification air units, and implemented various technical measures to reduce the bottle contamination rate. By using alcohol spray to reduce dust, the success rate increased to 92%, which has saved time in selecting the optimal culture medium formula and laid a solid foundation.

All equipment that comes into contact with culture media or can withstand high temperatures is sterilized using dry heat (140-150°C). For items that cannot withstand high temperatures, chemical sterilization methods are employed. To address the need for flame sterilization during inoculation (due to the lack of air inside the chamber), a 300-watt electric stove was added. However, to prevent damage to the explants from the high temperature, a ventilation and cooling system was also installed. To minimize secondary contamination of cutting tools, they are placed in a sealed tube after high-temperature sterilization, while clean air is continuously blown in to cool them down, ensuring the explants are safely transferred to the nutrient bottles. All items entering the chamber must be alcohol-disinfected before placement, and any item carrying bacteria is not allowed inside. The disinfection procedure for the chamber involves alcohol spraying followed by ultraviolet sterilization, and regular fumigation with formaldehyde and potassium permanganate.

⑶ Select appropriate culture medium formulations. Both induction and differentiation to rooting use MS basal medium supplemented with naphthaleneacetic acid and cytokinin. However, adjustments in composition are necessary at different growth stages, particularly during rooting. As orchids are more challenging to root compared to other flower varieties, a high-concentration NAA soak for a few seconds is employed, yielding good results.

④ Collection and sterilization of explants. The explants, being seedlings with long roots (unexpanded leaves), should be collected and inoculated immediately to achieve a higher survival rate (the best time for inoculation is from October to February of the following year). For the sterilization of explants, the author has found that using 0.1 liter of mercury and 0.1% concentrated hydrochloric acid for multiple sterilizations yields better results.

⑸ Cultivation Environment: Light intensity of 1000-2000 lux, 10-16 hours daily, cultivation temperature of 26°C ± 2°C, air temperature should be kept at a lower level (70%-80%), otherwise re-contamination may occur.

① Establishing a small tissue culture room at home is feasible; it requires minimal investment (fully utilizing home conditions). From preparation to conducting experiments, it only costs a few hundred yuan, and the experimental equipment can also be used for research on other varieties (such as woody plants, medicinal plants, etc.).

② Sterile operation must be taken seriously from the mindset and carefully executed; otherwise, it could result in heavy losses and unimaginable consequences.

③ Without a physical microscope, experiments can only be conducted by芽生芽. Implementing measures such as staged cutting of samples and the liquid-rotating bed method is a way to obtain large quantities of bulbils, and it can also break through the繁殖 speed barrier.