How To Successfully Grow Orchids From Seed Using Tissue Culture Techniques!

Today I will show you how to flask orchid seeds. Hi, my name is Anna and I am an intern at Plant Cell Technology. I'll be showing you how to grow orchids from seeds and tissue culture. Did you know almost all orchids you find in stores start off in tissue culture? The reason why is because in nature orchids rely on mycorrhiza, which is the fungus that facilitates the absorption of nutrients to the plants. However, those conditions are hard to replicate, which is why orchids are grown in tissue culture. So let's get started and grow our own seeds. I begin with pod sterilization, where I start off by using 70% alcohol to sterilize my workspace. I will then create a 50-50 mixture of both bleach and water that I will add to my jar. Next, I will add one drop of polysorbate 20 to act as a surfactant. This is the seed pod I'll be using, which I'll add to the bleach solution. Next, I will shake the jar and put it on our orbital shaker for 20 minutes. Now that the time is up, I'll re-sterilize my workspace and wipe down all surfaces. I will then open the jar with the seed pod and dump out the bleach solution, making sure not to pour out the pod. Next, I will add sterilized water to the jar and put on our orbital shaker for five minutes. I will pour out the solution and repeat these steps for three rinses total. After the last five minutes in the shaker, I will re-sterilize my workspace. I'll open the jar and pick up the pod to dip in 70% alcohol for a few seconds. Then, I'll place the pod on a glass dish and take a lighter to catch the seed pod on fire for further sterilization. I will get my tools ready and change my scalpel blade to split the pod in thirds. Then, I'll sterilize my three test tubes and fill each halfway with sterilized water. Now, I'll split the seed pod in thirds, making sure to keep the seeds intact. I will add each to a test tube with sterilized water. I noticed that even after agitating the seeds with a vortex, they would remain attached to the pod. I decided to cut the seeds from the pod with my scalpel and add them directly to the test tubes. Some of the seeds became separated, so it is okay if you have to add multiple clumps to the test tube at a time. I repeated this process for the remaining seed pod pieces and mixed each using the vortex. I noticed that the seeds finally started separating, so it is time to transfer them to media, which is very easy to make. You will first add 250 milliliters of distilled water and use a stir bar to agitate the solution in the media container. Next, you will add 6.84 grams of orchid media. I added 250 microliters of PPM to the mixture. After, I calibrated the pH so that it reached 5.8. The last step is to add 2 grams of agar to the solution, and I did this process twice for two media containers so that it could fill all 21 test tubes. Make sure to add autoclave tape on your media containers when you are finished. You will add both media containers and test tubes into the pressure cooker. Screw on the lid and don't forget to press start. 25 minutes later, first open the valve to release extra pressure and take out media containers and test tubes. Once you are finished, it is time to pour the media. Re-sterilize the workspace, including the test tubes and media containers. I chose to pour the media five test tubes at a time, and make sure to shake the media container before pouring because some charcoal may have stuck to the bottom of the glass. Repeat this process for all 21 test tubes, and make sure to shake the media container before each pour to agitate the charcoal. Now that the media is ready, it is time to add the seeds. I begin by re-sterilizing my workspace and using my micropipette to add 1.5 milliliters of the seed mixture to each test tube. Make sure to first shake the test tube using the vortex and suck in and out the seeds with the micropipette before taking 1.5 milliliters to ensure the seeds are thoroughly mixed. Repeat for the remaining test tubes and don't forget to mix the seeds between each tube. I had some seeds left over, so I decided to put the rest in sterilized petri dishes. I added 1 milliliter to each dish and even had some left over, so I added an additional milliliter to some of the dishes. Once I was finished, I put saran wrap around the petri dishes to prevent contamination. I also added barcode labels to all the dishes and test tubes so we can document the seed growth. Orchid seed germination can take a couple of months, so be patient. These are the finished test tubes and plates. This was actually my first time putting orchid seeds in tissue culture, and this was just one of the skills I've learned as a PCT intern. Make sure to check PlantCellTechnology.com for all your plant tissue culture needs and follow us on social media. Bye! Thanks for watching the video. If you are enjoying these tissue culture videos, make sure to follow us on social media for more informative content. And if you are interested in conducting your own tissue culture experiments, make sure to check us out at PlantCellTechnology.com for all the products you need to get started. Use code FP10 for 10% off on your first purchase.