Week 5

This was a very productive week for me in lab. Last week I finally got the dyes I was waiting for and I made agar for the first time in my life so I'm finally ready to start plating plant tissues in an attempt to isolate endophytic fungi. I plan on using three species of plants: lantana, oleander and bougainville, and three tissue types: leaves flowers and stems. 

Endophytes are slow growing fungi and if the surface of the leaves or flowers I'm using have any sort of fungal spore or bacteria on it then those organisms will grow much faster than the endophyte, take over the agar and eat all the food before the endophyte has a chance to appear. To ensure that the only thing that is growing in my agar is an endophyte, I have to sterilize the surface of the tissues. 

Monday I came in early to design the sterilization method I will be using on the plants for my project. The materials are first rinsed in running tap water for 5 min to remove any dirt or debris and then cut into 5mm lengths. The sections are agitated in 100% isopropyl alcohol for 2 minutes and then placed in a 1.25% bleach solution. At the intervals of 0,1,5, and 10 min four sections of tissue are removed, triple rinsed in sucessive sterile water baths and placed on an agar plate. It will take more than three weeks to see any results.