Tissue Culture Invitro Propagation
Along with convention propagation methods to increase nursery stock, our collection of plants is primarily propagated through tissue culturing and seed flasking. We specialize in a diverse range of plants, including bromeliads, air plants, staghorns, Pinguicula, and cacti. We are one of the few groups in the United States, and possibly internationally, with such a large collection of air plants grown in vitro. Growing plants in vitro allows us to rapidly multiply and expand our inventory compared to traditional methods.
We use two main techniques to cultivate plants in vitro: tissue culture and flasking. In tissue culture, small pieces of plant tissue are used to produce new plants. This method is ideal for creating large quantities of genetically identical plants, which is common in commercial production. Flasking, on the other hand, involves sterilizing seeds and placing them in a nutrient-rich gel. This technique produces a diverse group of plants (known as a grex), which is perfect for plant breeding and enhancing genetic variability.
While both processes are labor-intensive, they are highly rewarding when done correctly.
Two key factors in successful in vitro propagation are the media and the sterilization of plant material.
Media refers to the nutrient-rich gel in which the plants grow. It contains macro- and micronutrients, vitamins, sucrose (sugar), and agar. The exact formulation varies depending on the plant species, but agar is typically used at a rate of 7 grams per liter. The pH of the media is generally adjusted to 5.8. Once the ingredients are thoroughly mixed, the media is sterilized by autoclaving or pressure cooking to eliminate any potential contaminants. After sterilization, the media is poured into petri dishes, test tubes, or deli containers, and once cooled, it is ready for use.
Sterilizing the plant material is another crucial step, requiring careful attention. A surfactant like hand soap or Tween 20 is commonly used to coat the plant's surface, ensuring that no air bubbles or pockets remain where bacteria or fungi could thrive. The plant material is then sterilized with a chemical disinfectant—often a 10% bleach solution—for a period of 20 minutes to an hour. After sterilization, the plant material is rinsed with sterile water to remove any remaining bleach residue. It's important to note that certain species, such as carnivorous plants, may be sensitive to this level of treatment and can be damaged, so research on each species' sterilization requirements is essential.
Failure to properly sterilize the plant material can lead to contamination, which may result in the death of the culture and pose a health risk. Proper sterilization is essential to maintain healthy, thriving in vitro plants.