Lab work for the Week
Monday was slow I headed into lab bright and early in the morning to find that my advisor was working from home and that my fellow intern also would not be in lab. So I decided to pour some plates from the previous week and take photos of out antagonistic plates to right down some observations in my notebook. after ending early on Monday, Tuesday finally had some work for us to get done. This included going out into the filed and collecting some root and leaf samples. We collected from rice patties with a nitrogen value of 0 as well as a high nitrogen value patch. We collected 20 leaf samples and four root samples which we would process the next day. We would pull one plant and cut 5 leaves off of the plant and place them into a sterilized tube. And for the roots once the plant was one of the ground we had to shake/pull the mud off of the roots and place them into a 100 ml conical tube. When collecting the roots the area in which we will be isolating from is the rhizosphere, this is the area right outside of the roots (shown below).
This is where we are most likely to find endophytes that grow in the root systems. The next day on Wednesday we had the task to disinfect and plate the leaf samples. The procedure I found during my research to disinfect the leaves and roots was as the following:
- Rinse with sterile dH2O
- Dip in 7% ethanol for 1 minute
- Dip in 2.5% Sodium hypochlorite (Chlorox) for 4 minutes
- Dip in 7% ehtanol for 30 seconds
- Rinse three times in sterile dH2O
The set up or the disinfecting and plating process looked is shown below.
After the disinfecting process we had to cute our 4 leaf pieces into 2 and place them on two PDA plates and 2 NA plates. During plating we also had to injure the leaves by making small incisions on the sides to induce infection! Once that was completed which took the entire day by making 40 plates each, we placed the NA in the 37 degrees C incubator and the PDA plates in the 28 degrees C incubator. Thursday was yet another busy day filled with mainly making plates for the next day of experiments. We made 40 plates of both NA and PDA as well as about 40 tubes of PDA slants and NA Slants. Friday was a bit more exciting with Vincent returning and us being able to see some of the cable bacteria that grew in the slides for a week!
The cable bacteria quite literally look like cables or to me they look like hair under the microscope! These do help with reducing methane emission in rice soils however, they do make the soil quite acidic with the amount hydrogen they produce.
next we had to analyze our results from our leaf plates! Most of them seemed to grow either a fungus (on the PDA) or a bacteria (on the NA) below are the results!
We did not have enough time in lab this week to streak for isolation but we will complete that task next week.
The last activities we did on Friday is go out into the field again with Vincent to collect sample from the high nitrogen levels patch. The images of the cable bacteria came from a soil sample of high nitrogen. The samples from low nitrogen had no cable bacteria. So we scooped a huge bowl of mud and sifted the mud using a strainer and then put it in a conical tube! It was very hot that day and we were all sweating quite a lot, so after sampling we all went for an iced coffee to end our work week!
This was another slow weekend. We started it on Friday night where we went to a Karaoke place called LBeats. We sang so many songs, its was fun! Then on Saturday the group split up with me and Julia hanging around Los Banos due to an errand I had to run. The idea was that we needed to find Bayluscide which is an apple snail control that is only sold here, and my dad wants some for our taro patches back home. When I tell you it was journey getting there, it was. We started at a café to get some food into our system. I had some waffles and a brown sugar latte, Julia got a strawberry latte and we both thought it was quite interesting. Next we went to find a charger because Julia’s phone was about to die and she had the google maps on it. After that we headed into a part of town that we both haven’t walked in. The streets were busy and after another 20 minutes of walking we stumbled into the small farming supplies store. The store was so small it seemed as though it could only fit three people. I was lucky enough to get the last box of 10! Next we decided to hang at one of the smaller malls where Julia and I sat at a coffee shop reading our palms for like 2 hours. Then we decided to get some messages in the message chairs here.
Once Julia and I finished our we decided to start the trek back. After walking for a bit we were hungry so we stopped at a somewhat fancy restaurant called Bonitos. It felt so nice to sit down in an AC room for a bit, plus we were very hungry. We got our food and enjoyed our time. When we were about to leave there was a huge crack of thunder followed lighting and some heavy rain. We were stuck in the restaurant for a bit and then when the rain lightened a bit we made the decision to head out. It was drizzling a bit but the thunder and lighting were heavy. It was quite beautiful though because we were walking during sunset so when the lighting flashed the whole sky turned purple. Sunday was yet another lazy day where Jessica N. Julia and I went to a café to do some work and had some yummy treats. Till next time folks! 🙂