Tissue Culture Beginner's Guide - Learn ALL the Basics

Summary

In this tutorial, the creator walks viewers through the entire process of plant tissue culture, teaching how to clone plants at home. The video covers essential supplies, including tissue culture media, ingredients like distilled water, MS, sugar, jelling agents, and plant growth regulators. It details the preparation of the media, sterilization, and transferring plants into fresh media to promote growth. The tutorial also includes tips on avoiding contamination, selecting plants, and ensuring success with tissue culture. Additionally, the creator highlights helpful resources and a sponsored tissue culture masterclass for further learning.

Takeaways

• 😀 The video teaches how to start with plant tissue culture at home, cloning plants from your own space.
• 😀 Tissue culture media provides essential nutrients, vitamins, and hormones for plants to grow outside of their natural environment.
• 😀 The five key ingredients for making tissue culture media are distilled water, MS medium, sugar, a gelling agent, and plant growth regulators (PGRs).
• 😀 Different plant protocols require different media recipes; they can be found on platforms like ResearchGate or Google Scholar.
• 😀 The correct pH range for tissue culture media is 5.6 to 5.8, and pH adjustments are made using hydroponic pH up/down solutions.
• 😀 Sterilization is crucial for tissue culture to avoid contamination; the media and tools like forceps should be sterilized in a pressure cooker.
• 😀 The best way to avoid contamination when starting tissue culture is to use plants already in tissue culture rather than those from traditional soil-based growth.
• 😀 Starting with plants already in tissue culture helps ensure a sterile beginning for the cloning process, reducing the risk of failure.
• 😀 A still airbox or laminar flow hood can be used to maintain a sterile environment while transferring plants into multiplication media.
• 😀 Regular subculturing every 4-6 weeks is necessary to prevent buildup of ethylene gas, which could kill the plants in tissue culture.
• 😀 Once the plants have multiplied enough, they can be transferred to rooting media or acclimated to soil to complete the process.

Q & A

What is the primary purpose of tissue culture media in plant cloning?

Tissue culture media provides all the necessary nutrients, vitamins, and hormones for plants to grow outside their natural environment. It helps facilitate the cloning process.

What are the five key ingredients in tissue culture media?

The five key ingredients in tissue culture media are distilled water, MS (Murasige and Skoog) basal salt medium, sugar, a gelling agent (like agar or gelling gum), and plant growth regulators (PGRs), which are plant hormones.

Why is it important to maintain a specific pH level in tissue culture media?

Maintaining the pH between 5.6 and 5.8 is essential because it ensures that the plant can efficiently absorb nutrients. Deviations from this range could harm the plant's ability to grow and develop.

How can tissue culture media be sterilized?

Tissue culture media can be sterilized by pressure cooking it at 15 psi for 15 minutes. The containers should not be sealed tightly before sterilization to prevent pressure buildup, but should be sealed immediately after opening the pressure cooker.

What is the recommended method for sterilizing forceps used in tissue culture?

Forceps can be sterilized by autoclaving them in a pressure cooker for 15 to 20 minutes at 15 psi, just like the media.

Why is using plants already in tissue culture recommended for beginners?

Starting with plants already in tissue culture ensures that they are sterile from the beginning, reducing the risk of contamination. This allows beginners to focus on the multiplication process rather than dealing with sterilization challenges.

What are some tips to avoid contamination during the tissue culture process?

To avoid contamination, work quickly, avoid leaving containers open for too long, never reach your hands over open containers, and use sterilized forceps and tools. Additionally, use a still airbox or laminar flow hood to maintain sterility.

What should you do when you notice contamination in your tissue culture?

If contamination occurs, such as with the Monstera in the video, discard the contaminated plant and start over with a new, sterile plant. Contamination can ruin the entire culture.

How often should you subculture plants in tissue culture?

Plants should be subcultured every 4 to 6 weeks, which involves transferring them to fresh media. This prevents the buildup of harmful ethylene gas and ensures continued growth.

What are the next steps after successfully growing plantlets in tissue culture?

Once enough plantlets are formed and the media is filled with shoots, you can transfer them to rooting media or acclimate them to soil if they are large enough. These plantlets can then be moved out of tissue culture for further growth.