Weeks 3 & 4: Final Trial Preparations

Hi! Welcome to the third and fourth weeks of my project blog! 

The past two weeks have been primarily remote because my project advisor (Matias) was traveling for a scientific conference. On the bright side, Mrs. Baessa and I had plenty of supplies available to work with, and extra time at Genspace allowed me to gain some insight into a possible agrobacterium transformation for alfalfa. 

After visiting CSHL the past week, I determined that testing the two soil groups prior to planting would be ideal. With nitrogen compound testing strips, I tested the available MGS-1 simulant for its nitrate and nitrite concentrations. As predicted by the simulant spec sheet, nitrate/nitrite levels in the soil were shown as zero (Figure 1). So in the following growth trial, soil will be collected and subsequently tested to determine the amount of nitrogenous compounds produced by the alfalfa root system, Azospirillum fertilizer, or both. 

Additionally, previous meetings have drawn my attention to creating a conditions sheet for alfalfa in order to determine the best environment at CSHL for running the final trial next week. My research was rooted primarily in humidity and temperature, because the laboratory separates several chambers based on their sterility, humidity, and temperature. Meanwhile other factors such as soil depth, pH, level, and watering may be adjusted accordingly by the users preferences. Ultimately I determined that experimental groups will require full sun, humidity of approximately 60-70%, temperatures of 60-65 degrees Fahrenheit, and a randomized block layout to grow most efficiently given the circumstances of my research. A randomized block design refers to a system of distributing experiment groups completely randomly within a given space in order to reduce result biases due to uneven conditions; given that the greenhouse/growth chambers that I will use may have varied conditions across the different flats I use, next week I will randomize the placement of several hundred plant trays. 

Finally, to end the week I visited Genspace to continue working with the Open Plant Community project. With several other members, I transplanted common liverwort (also known by its scientific name Marchantia) from agar plates to larger boxes for long term storage (Figure 2). Our propagations included several strains of Marchantia, namely Tak1, Cam1, and RUBY – a red variety cultivated by inserting a plasmid that expresses the red pigment betalain (commonly observed in beets). While alfalfa and liverwort only share the title of “model organism,” agrobacterium affects them equally. I hope to pursue producing transgenic cultures of alfalfa with Matias’s plasmids in the coming weeks, and look forward to documenting differences between Marchantia Polymorpha and M. Sativa transformation. 

See you then!