Tissue Culture Experiment 2

Following on the results of my last experiment, I plated up a few more samples of D. Muscipula (venus flytrap) seeds in media jars, this time with added nutrients. I wish I had kept better track of what I added to the media, but I got distracted.

Flytrap seedlings in tissue culture media. I added food coloring to keep track of different concentrations.
Flytrap seedlings in tissue culture media. I added food coloring to keep track of different concentrations.

I’ll speculate that the above specimen has approximately 1/8 cups sucrose per Liter, a pinch of generic plant fertilizer, and a dash of multivitamin. This was the most dilute sample I made. Plates with higher sucrose concentrations got contaminated and had to be thrown out.

I was too stingy with the agar and the media failed to solidify. I hoped the seeds would float and be happy, but they promptly sank to the bottom. I left the jars on a windowsill, assuming they were a lost cause.

Surprisingly, the seeds germinated while completely submerged. After the leaves grew out, the seedlings floated to the surface. Despite the fact that it is the middle of winter and the plants are receiving no supplemental heat or light, they are still successfully producing chlorophyll by absorbing sugar and trace micronutrients from media.

Growing plants have greater nutrient requirements than seedlings. However, I am loathe to replate these guys for lack of a sterile work environment. I’ll see how long they last in the current media and check back later. I am happy with how things are progressing.

For my next trick, I’ll slice off a piece of my arm to grow in vitro.

Tissue Culture Experiment 1

Man, having a job really infringes on my plans for world domination, no matter how much I intentionally suck at my job. This update is long overdue.

This all began a few months ago, when I decided to learn to do tissue culture so that I could ultimately clone myself. I started with plants, since plant tissue is easy to work with, and I have had a lot of practice with, uh, herb gardening. My first attempt at tissue culture succeeded only in growing plates of mold. I threw those out, although I suppose I could have harvested some penicillin.

For my next attempt, I put some venus flytrap seeds in plain agar with no additives. I was still quite lax with my sterilization techniques, but none of the plates were contaminated this time because there was nothing but agar.

Newly hatched flytraps in agar

I moved one plate of seeds to peat moss shortly after germination because the agar was drying out. The other plate remained for about a month after germination. I finally moved the last plate to soil because the specimens were looking sickly.

The plant in the middle and on the right were moved to soil shortly after germination, and the plant on the left was in tissue culture until one month after germination. Obviously, I didn’t do it right.

There were a number of things that I did wrong with my tissue culture.

Sugar: I had originally thought that adding sugar to the agar would be stupid because, wtf, plants don’t eat sugar. But one reason why the ex-vitro plants did so much better than the in-vitro ones was the availability of light. Plated samples cannot be kept under direct sunlight or they’ll cook. Thus, plant tissue cultures do not rely on photosynthesis as an energy source. Sugar must be added to the growing media — apparently, plants can absorb and transport sucrose. (This doesn’t work well with ex-vitro plants because the bacteria and ants will get to it first.)

Venting: I used those swine-flu-epidemic surgical masks as lids for my cultures. Keeps the germs out, but still allows airflow. Not necessary, and allowed the agar to evaporate quickly. If photosynthesis is not transpiring, then there will be minimal need for gas exchange. Cleaner to use a closed lid.

Replating: The seedlings should have been replated after the first traps started forming. The nutritional requirements of a growing plant are different from those of a seedling which is still deriving nutrients from the seed.

Other additives: Thiamine is essential to carbohydrate metabolism, macronutrients such as N, P, K as fertilizers, plant growth regulators like cytokinins, and other stuff but I stopped reading because I got bored [1].

I’m going to try one more time.

1. Trigiano, R. and Gray, D. Plant Tissue Culture Concepts. CRC Press, 1999.

Tissue Culture Experiment 0

I made my first attempt at tissue culture over the weekend. Nothing too elaborate, just trying to establish a good control system for future experiments.

Here I’m trying to grow some squash in a medium made of agar and K-mart plant fertilizer. If this goes well, then I’ll try to clone something more exotic, like wasabi. If that works, then I’ll cut off a piece of my arm and use that for my next tissue culture. Then, world domination.