After a long time of talking, we finally put a plan into action, leaving the world of theoretical microbiology behind and entering the world of practical application.
The first part of our long journey (to be outlined at a later date) was to select some plant samples and sterilize their surface. Sterilizing their surface with a 95% ethanol solution, 3% sodium hypochlorite (bleach) solution, and 70% ethanol solution hopefully ensured that all bacteria,fungi, or spores on the outer surface were destroyed.
Our goal is to cultivate what is on the inside, endophytes. Symbiotic microorganisms that live within higher plants and produce a wide variety of bioactive compounds.
Shown here is one of our sample organisms, Ginko biloba, a “living fossil” whose leaf structure looks unchanged after almost 270 million years, quite an extraordinary organism!
We chose a few other plant specimens and put them through the same surface sterilization protocol. After washing with sterile water a few times, small pieces of plant tissue were excised and plated on potato dextrose agar (PDA) plates in the center of the plates. We also made sure to make a few control plates which involved simply pressing the sterilized leaf surface on the PDA, such that anything that grew on that plate would most likely be from the outside of the leaf, and if it showed up on any of the experimental plates, it’s roll as a strict endophyte would be put into question. This was also a good way to test the efficacy of our sterilization protocol.
After pieces of stem and leaf were plated, they were placed into an incubator of sorts at 25Cº +- 2Cº. I say “an incubator of sorts” because we don’t have access to a real laboratory grade incubator, so instead we used an old offline oven which had nice seals and minimal air flow and kept at a steady 23ºC all day.
An update will come later this week as they have been incubating all weekend, and on Monday we will get to see if anything has grown. Until then, here are some pictures of some of our samples already plated.