*This post is part of a series of posts based on my research and experience as a masters student at UGA. To find out more go to my page Holy Basil Research.
I believe most grad students go through this experience. You start your research project with big ideas and expectations of what you want to learn, as you go through you encounter challenges and limitations that affect your project and it evolves over time. Then, all of a sudden you look back and realize your research question is entirely different than when you started. At least this has been my experience.
When I was helping to write the grant to get funding for my masters research we had a clear research question. We wanted to see if we could manipulate growing practices to increase the quality of holy basil through the quantitative measures of yield, antioxidants and essential oils. I received the funding, and once I started grad school we had to come up with a plan of how to do that.
We realized there were many different varieties of holy basil and didn’t know which ones to grow for our study. So we acquired 14 different varieties; 5 from Horizon Herbs Seed Company, and 9 from the USDA seed repository in Ames, Iowa (read more in my post about seeding holy basil). We thought we would do a preliminary screening of these 14 to see which ones performed the best so we could choose the top 3-5 to do a cultural practice study. It is good to be thorough, right? We thought we could do this preliminary variety trial during my first summer/fall semester and use that data to choose holy basil varieties to run two cultural practice studies in the summer of 2016. For the cultural practices, we wanted to look at the effect of pinching back the flowers and cutting back the plant to encourage branching. Did it increase yield? Did it increase antioxidants and essential oils?
However, as we continued the variety trial with our 14 different kinds of holy basil we found that there were major physical differences. They had different coloring, growth habit, leaf shape and texture, flower size and time of flowering. We wanted to explore these differences further. I have included a picture and name or accession number for each of the 14 varieties in this post so you can see the differences for yourself!
When I gave my Departmental Proposal Seminar I was given feedback about the need for repeating the study. I couldn’t publish my results from my variety trial with only 1 year of data. Getting published is an important part of research, so I began to realize that I couldn’t just view this is a preliminary screening anymore. This variety trial had to be completely replicated next year if I wanted to report any of my results.
So, we put the idea of a growing practice study on the back burner while we poured our attention on the variety trial. We moved forward on building the protocols to extract the essential oils. After building the protocol, we realized that each distillation would take 3.5 hours. Since repetition is important to determine statistical significance, we had 3 reps of each of the 14 varieties over 2 seasons of harvest (read more about the reason for repetitions here). That meant I had 84 samples to distill. That is 294 hours of lab time just to get the samples of essential oil. This doesn’t include time to analyze them or run statistical analysis. So this project is turning out to be bigger and more involved that I originally planned.
Once I I finish distilling the samples, I will run them through the Gas Chromatograph Mass Spectrometer (GC/MS) to identify the compounds in each sample. For example, it would tell me that the kapoor holy basil has 60 different chemical compounds and one of them is the compound eugenol. Then I will run the sample through a Gas Chromatograph Flame Ionization Detector (GC/FID) to determine the relative amounts of each compound in the sample. For example, it might tell me that the concentration of eugenol in that sample of kapoor holy basil essential oil is 1.8% relative to the other 59 compounds in that sample. These are just examples. I haven’t done any of this analysis yet.
Once we have collected all this data. The mass of the dried holy basil, the mass of essential oil that is produced by each plant, the composition of each essential oil and the relative amounts of each compound in each essential oil. Then we will do our best to perform a statistical analysis so we can choose the top performing holy basil varieties.
Then the plan is to replicate this entire experiment next year. We will plant seeds from all 14 varieties, we will grow out the same amount of plants, plant them at approximately the same time as last year, get two harvests, etc…
This will allow us to compare the results from both years to see if we keep the same top performers and run the more in-depth analysis for a second year on those, or see if the list changes based on the second year of data.
In all of this, I still haven’t made any headway on figuring out what to do for an antioxidant analysis on all of these samples, and since we have to redo the variety trial experiment I likely won’t be able to do any cultural practice study. This has been an eye opening experience for me. Coming up with a great research question is only the tip of the iceberg. As the project progresses there are countless limitations of time, budget, resources, accepted standards in the field, opinions of different professors, good study design and having to back up every research decision with a previous standard or logic already published in the literature.
The reality is that many research projects change over time. Boundaries have to be created. Expectations have to be revised. There may be disappointment. But it can also create new opportunities, build problem solving skills and encourage flexibility. I feel like I say this a lot, but I am really learning so much from this project. I am sure things will continue to change, and the plans presented in this post will be different than the reality of what happens as I move forward in my project. But that’s okay. This is how science moves forward, one step at a time.
If this interests you and want to find out more, go to my page, Holy Basil Research!