In fall 2022, we planted Echinacea within our study site for the seed addition experiment. In the lab, we are currently doing a germination experiment with the same batch of seeds to calculate the expected seed germination rates under ideal conditions. Following our standard germination protocol, we first treat the achenes with Florel solution and put them in a cold fridge with low light levels to mimic winter conditions. After 14 days, we transfer them to a warm, bright environment to germinate.
Jared applies FlorelGerminating EchinaceaLindsey plants Echinacea
The seeds generally don’t start to germinate until they are in the warm environment. This year, however, I checked on the seeds a few days after they went into the fridge and discovered that they were already starting to sprout! The condenser on the fridge had broken, so it was no colder than room temperature. In spite of the unusual conditions, ~80% of the seeds germinated. This is very encouraging since it means that the seeds that we planted in the fall were viable.
After fixing the fridge, we are germinating a second batch of seeds following the standard protocol so we can replicate the experiment next spring. Lindsey planted the seeds that germinated, and we plan to grow them into plugs and transplant them this summer.
One of our main goals over the past two years has been to process all of the Echinacea harvested from remnants in 2020, 2021, and 2022 to investigate the effects of prescribed fire on flowering and fitness. We harvested 1,012 heads over these three years.
year
# of heads
rem2020
299
rem2021
383
rem2022
330
For each head, the end goal is to get an accurate count of the number of achenes and the seed set, a measure of pollination success. To collect these data, we sent the Echinacea through our high-throughput seed processing system, which we call the ACE process. (The true meaning of ACE is controversial: Always Cleaning Echinacea? Accurately Counting Echinacea? Achene Chaos Extraordinaire?) The ACE process has many steps: inventory, clean, recheck, scan, count, randomize, x-ray, classify, store.
In spring 2023, we finished processing all of the remnant Echinacea from 2020, 2021, and 2022! We send a huge thank you to all the students and volunteers who put in many hours on this project over the last few years. We couldn’t have done it without you!
Luk and Sue cleanWanying and Caitlin recheckLake Forest College students recheck and scanKat scansAllen countsPriti randomizesMarty, Char, Padmini, Caroline, and Allen agree: it’s a team effort!
Jared is currently adding the achene count and seed set data to the remnant data repository on the Echinacea Project website. All achenes are currently in the dehumidifier at the Chicago Botanic Garden. After two weeks of drying, they can be stored in the seed bank.
I know it has been a while since I last gave you all an update, but we have some good news to share! We managed to finish cleaning the third batch and are close to completing randomization for the second batch. I will say our progress has been pretty steady before the last update I shared with you, but we decided to out some ways to accelerate processes, such as randomization. We created an assembly line (shown in the image above). We tasked people to either set up the stations by pouring and spreading out the achenes onto the randomization grids or randomly select grids in which achenes will be inspected for predation. We found this setup to be much more efficient, and hopefully, by the end of next week, we may have three batches fully cleaned and randomized. However, our initial goal of completing five batches may be somewhat too ambitious, so instead, we decided to just focus on three batches so there may be time to collect and analyze the data. In the figure below, you will find our overall progress made on the project, and given that my focus would be on studying predation rates on Liatris plants, the stuff that comes after randomization does not necessarily apply to the findings I will need. With that said, those other sets of data will most likely be of use for other studies, and the Echinacea Project will most certainly take a look into those things. Overall, things are looking good for us with the project, and I cannot wait to see the results soon.
P.S., I did not have a chance to specifically address my project’s intentions in an ABT prompt, so here it is: “We know that Liatris plants get eaten more often than Echinacea plants, and predation rates tend to be higher in the absence of fires, but we do not know which types of which types of Liatris plants may be more likely to be eaten than others. Therefore, we are studying how the number heads of a Liatris plant affects the predation rate on the plant’s achenes.”
Pollinators are declining worldwide, a phenomenon that some people are calling the insect apocalypse. There are many factors driving these population declines, and loss of habitat has been identified as one major cause of insect demise. In our study area in western Minnesota, we have seen numerous prairie patches converted to agriculture over the years. However, we don’t know how the bee community has changed over time across the landscape. To investigate these questions, the Echinacea Project started the Pollinators on Roadsides project back in 2004, and we collected another year of data this past summer. In the fall, we brought 7 coolers of insects back to the lab at the Chicago Botanic Garden.
This winter and spring, volunteer Mike has been busy as a bee pinning all the specimens so we can send them to Zach Portman, a bee taxonomist at the University of Minnesota. This week, Mike started working on the last cooler of bees! So far, he’s pinned 680 insects collected in 2022.
The 610th bee from summer 2022Mike shows off the 2022 bee collection
Funding for this project was provided by the Minnesota Environment and Natural Resources Trust Fund as recommended by the Legislative-Citizen Commission on Minnesota Resources (LCCMR). The Trust Fund is a permanent fund constitutionally established by the citizens of Minnesota to assist in the protection, conservation, preservation, and enhancement of the state’s air, water, land, fish, wildlife, and other natural resources.
We made it to the third batch for the cleaning procedure! After completing the first batch for randomization last week, we needed to pick things up on cleaning, and we hit another promising milestone. Ultimately, we wanted the cleaning process to stay well ahead of the randomization process, and we did manage to get ahead of ourselves some on randomization last week. We made much more progress in that department than I had envisioned, and we caught up a little too much concerning cleaning progress. We focused on cleaning all this week, and now here is where things stand. There is still much more to go, but we are steadily getting there.
Good news! We managed to get through randomizing the first batch! Now we are working our way through the second batch, and so far, things have been going pretty well in terms of pace and efficiency for both cleaning and randomization. Overall, we have seen a considerably wide range of head counts in the Liatris plants, but predation rates have been relatively low compared to what was observed in 2021’s data. Who knows what we will find in the next several batches, but I am pretty excited about the progress made so far and the results we may see. Hopefully, we will complete the next batch within the next few weeks, if not sooner.
So far, things are going well in the randomization step, but there is still quite a ways to go to complete the first batch of Liatris plants. However, we have counted a considerable number of achenes that have been eaten, and we are starting to get a rough estimate of the predation rate per Liatris plant. We are finding that there are, on average, 1 to 3 achenes eaten per plant out of a total of at least 30 randomly selected achenes. This equals approximately a 3 to 10 percent range for what has been found, but we have also found a few that have had either none or many achenes eaten. In light of these findings, I think we are heading in the right direction with the project, but hopefully, we will need to get much more work done to get through all five batches. Hopefully, we will have all the plants cleaned and randomized before the burning season (which starts in April), but it will take a lot to get to that goal within a little over a month’s worth of time. However, the good news is whether or not we meet our goal, there will be more than enough data to work with, and ultimately we can come to conclusions at the end.
In lab today, I started the process of randomization. This step in the project does take some time and patience, but all of it is worthwhile toward reducing bias and collecting achenes that can be x-rayed or identified as eaten. Similar to the randomization of achenes from Echinacea plants, sheets of randomization and counting grids are used. In addition, we also use plastic bags, white envelopes, stickers, tweezers, and a magnifying glass for this procedure. The first step involves pouring the achenes evenly across the randomization grid and ensuring all the achenes are placed in a specific square on the grid without being on the lines. Then using a randomized list of grid coordinates, we go down the list until we hit a coordinate (an example would be “B2”) with achenes present. We then examined each achene in the selected coordinate to see if there was any predation. If less than 30 achenes were counted in the coordinate, we randomly selected another one going down the randomization list. Once we categorized 30 achenes as either eaten or eligible for x-ray, we took our white envelope and plastic bag and put stickers on them. The envelope will store the eaten achenes while the plastic bag will contain the achenes ready to be x-rayed. On the envelope, we write the date, initials, and the quantities of achenes eaten, not eaten, and uninformative. After putting the achenes in their respective containers, we put them in a pile marked “randomized,” and we move on to the next one. As of today, we managed to get through a little over one third of our first batch, so we still have much work to do before proceeding onward.
After cleaning Liatris plants for a week, I am happy to say that the first of five batches are now finished. Now that the first batch is completed, the project’s next step is ready to begin. The process of randomization will be the next step in the project. This step will involve the random selection of achenes from each plant to avoid bias and separation of achenes that are either qualified for being x-rayed or not. Luckily, both groupings will have a role in the project’s studies. X-rayed achenes will inform us about pollination and reproductive outcomes for each head of a Liatris plant. On the other hand, non-x-rayed achenes can be assessed for why they cannot be x-rayed, including what types of damage the achenes have and if any predation occurred toward those achenes. My research question will focus specifically on seed predation, so using the non-x-rayed achenes will be essential. I am still working on a finalized research question relating to seed predation, but seeing the progress made so far has me excited about what will come next in the project and toward finalizing my research question.
I am happy to say that the Liatris Project is off to a good start. After taking inventory of all the Liatris plants this past week, I got to start the cleaning process. A total of 293 Liatris plants have been counted in the inventory, and all have been sorted randomly into 5 different batches. Today, I got to start cleaning the ones in the 1st batch, and while cleaning, I noticed several similarities and differences compared to cleaning Echinacea plants. Overall, I found that Liatris achenes were much easier to extract from the plant than Echinacea achenes, but counting them proved much more challenging. To make things easier, random selection sheets of different numerical ranges were arranged that listed random numbers from left to right down the sheets. Using these sheets, I could randomly pick out a flower head and count the number of achenes associated with that head. I also had to observe if any achenes were missing from each head on a Liatris plant. I recorded the total number of heads per plant and the number of heads with no achenes, some achenes, or all achenes missing. After taking these recordings, I removed all the other achenes present on the Liatris plants and sorted them into an envelope. Any chaff leftover got put into a separate envelope labeled as “chaff.” So far, a handful of plants have been cleaned, but there is still a long way to go.
