Here is the fungus I am trying to isolate!

With the help of another intern (Amanda) this week I was able to actually find and identify the fungal endophyte growing in my grass. Neotyphodium is the genes of the endophytic fungi i am finding in the festuca grass samples I am testing on. Now the hard part is to isolate them. In some further research some scientist are using antibiotic enriched media to prevent bacterial growth but not effect fungal growth. Some antibiotics have no effect on a fungus because fungi lack the proper chemical receptors for the antibiotics to target - much like viruses. This may be the next step in finding a media to grow what I want. In the mean time here are some of the pictures of endophytes I took this week. If you look at last weeks post the blue lines are fungus growing between the plant cells and they are very prevalent in these pictures as well. 

Rought draght was rough

One of my agar's has continued to grow and is looking like it could be an Endophtye but I am not going to hold my breath. It looks like just colonies of fungi have grown and no other microbes seem to be present on the plate, which is the hardest part so far this projects, getting an isolated microbe to test on. The rough draft due this week was just that very very rough and incomplete. I am feeling the stress of not having enough conclusive data by the end of the semester to conclude my research and answer the question for my experiment. I think I have had many successes in the last few weeks but still nothing in the way of fungal growth. I am almost positive that one of the bacteria I was able to isolate is an endophyte but I was not trying to isolate a bacterial endophyte to begin with. Fungal endophytes are much more selective and hard to grow. I am going back to the beginning to see if the grass i am using in fact has fungal endophytes in it. I will use a process of staining extremely thin slices of grass with Anil blue to identify if the grass is in fact infected with fungus. If all goes correctly it should look something like this.
     

granola troll hole

Last week the pressure of the semester really started to weigh on me and then before I realized it the week was over and it was time for a Friday fun day. Field trip with the other interns to Dreamy Draw park for a little lesson on navigation, latitude, longitude, and coordinates. There was a very interesting talk on ephemeral drainage, and the conspiracy theory that alien crash sites have been covered up in those hills. I went to hide a geocache for the first time and think we found a great spot.

The project is going slow and steady but at least it is going. I am still trying to isolate and identify the microbes I keep inoculating from the grass. I am at the same time adjusting my sterilization technique and trying to get a smaller number of microbes per slide. Unfortunately the dichotomous key Josh and I found for MEA is not for the type of fungus I am trying to isolate. MEA is still a good media to use for the fungus I am isolating I just need to find another dichotomous key. 

Malt shakes any one?

To all of the hard working mentors and students in the program, have a great and happy holiday. This week in lab I have made yet another media to try and isolate these fungal cells. In my research I found an interesting article talking about endophytes actually giving plants tolerance to pollutants in the air, like from automobiles. I am amazed at how symbiotic these relationships are between endophytes and plants and just how much each one can gain from each other. The malt extract I am using is going to require 20 grams of malt extract and 9 grams of agar solution as a standard from the dichotomous key I am using.  
 

Here is the video from last week sorry guys.

I will have to admit even with all the stresses of life weighing me down, the time spent in lab and with the great minds that encompass that area has been a centering point for thought, laughter, and of course some science work. I am thankful for all the joy this class brings me. Now with that said I made corn meal agar and will make malt extract agar this week to go along with the new dicotomist key we found. That way I can see if the microbes I have isolated are in fact endophytes. These media should help differentiate and help isolate the fungus I am looking for. Untill next week here is a video for your enjoyment.

This is why endophytes are important!

Things seem to be taking turns very quickly. One day I am excited that I got something isolated and a pure enough sample to test on, and then the next day back at square one. I am going to try using different media's designed for fungus growth to see if that makes any difference. With the help of Josh (thanks again by the way) I was able to find a dichotomous key that will help me in identifying the microbes I am finding. They used MEA (malt extract agar) to inoculate there microbes as well as corn meal agar. Josh will be ordering these media for me so I can make some new plates and try again. Even though things are going slow, I feel I am making strides and improvements in the right direction. This video shows how endophytes have already helped the world and the problems of agriculturist everywhere.