In summer 2022, I continued the interremnant crosses experiment to understand how the distance between plants in space and their timing of flowering influences the fitness of their offspring. This experiment builds on my study of gene flow and pollen movement in the remnants, asking the question of how pollen movement patterns affect offspring establishment and fitness. If plants that are located close together or flower at the same time are closely related, their offspring might be more closely related and inbred, and have lower fitness than plants that are far apart and/or flower more asynchronously. In other words, if distance in space or time is correlated with relatedness, we’d expect mating between more distant or asynchronous individuals to result in more fit offspring.
To test this hypothesis, I performed crosses between plants across a range of spatial isolation (within the same population, in adjacent populations, and in far-apart populations) in 2020. With the team’s help, I also kept track of the individuals’ flowering time to assess whether reproductive synchrony is associated with reduced offspring fitness, suggesting that individuals that flower at the same time are more closely related.
In 2021, I repeated the same hand crossing methods to assess the fitness consequences of outcrossing on 44 focal plants. However, instead of planting the offspring from these crosses as seeds, I germinated them in the growth chamber and transferred sprouts to a plug tray.
In spring 2022, with help from the team, I planted the seedlings as plugs into ExPt1. I measured the seedlings throughout the summer.
Lindsey digs a hole for an Echinacea plugA baby Echinacea!Amy plants Echinacea in ExPt1 after the burn
To learn more about Amy’s project, check out this video created by 2021 RET participant Alex Wicker.
Start year: 2020
Location: On27, SGC, GC, NGC, EELR, KJ, NNWLF, NWLF, LF
Data collected: Style shriveling and seed set and weight from crosses, start and end date of flowering, coordinates of all individuals in the populations listed above. Leaf count and height of seedlings at three points during the summer (two weeks after planting, mid-summer, and late summer).
Beginning in summer 2021, we have been studying fire effects on Liatris aspera across patches of remnant prairie in Solem Township, MN. Six of these remnants burned in spring of 2021, and five remnants burned in spring 2022. During 2022, we expanded our efforts and collected data at five additional (smaller sites). The (absurdly) high density of flowering Liatris during summer 2022 led us to establish 1 meter wide, randomly placed transects in many sites. Despite our efforts and stated goal of not mapping as many Liatris as we did in summer 2021 (when we mapped 2400+ flowering plants across 23 remnant patches), we managed to overshoot our 2021 counts by at least 33%. Yes, we mapped well over 3200 flowering Liatris during summer 2022 with but a single functioning GPS… We harvested seed from 291 randomly selected Liatris.
Start year: 2021
Location: 28 patches of remnant prairie in and around Solem Township, MN
Overlaps with: Foolishness and shenanigans
Data collected: Demographic and spatial data housed in the remla Bitbucket repo
Samples or specimens collected: Harvested seed heads have been dried and are located in Jared’s office. These need to be inventoried, cleaned, randomized, scored for seed predation, X-rayed, and classified.
Products: Stay tuned!
You can read more about the Liatris fire and flowering project, as well as links to prior flog entries about this experiment, on the background page for this experiment.
In 2022, we collected data on the timing of flowering for 1373 flowering plants (2291 flowering heads) in 23 remnant patches. We identify each plant with a numbered tag affixed to the base and give each head a colored twist tie, so that each head has a unique tag/twist-tie combination, or “head ID”, under which we store all phenology data. We monitor the flowering status of all flowering plants in the remnants, visiting at least once every three days (usually every two days) until all heads are done flowering to obtain start and end dates of flowering. In most remnants, we monitor the phenology of all flowering Echinacea. In 2022, record high flowering led us to sub-sample from select sites (namely alf, nwlf, lfe, lfw, lce, and lcw) and exclude some sites (e.g., ri, aa, and a large chunk of alf).
Linking this detailed phenology data with information about seed production will help us understand how the timing of reproduction influences pollination and reproductive fitness. Additionally, we are excited to investigate whether fire synchronizes flowering in remnant populations. Eight of the populations in which we collected phenology data were burned during spring 2022.
Start year: 1996
Location: Roadsides, railroad rights of way, and nature preserves in and around Solem Township, MN
Data collected: We managed the data in the R project ‘aiisummer2022′ and will add the records to the database of previous years’ remnant phenology records. The 2022 phenology data set needs to be cleaned and prepared for integration with phenology data from previous years and is still located in the aiisummer2022 repo.
Products: Stay tuned!
You can read more about the Flowering phenology in remnants project, as well as links to prior flog entries about this experiment, on the background page for this experiment.
After a foolhardy effort to single-handedly census every flowering Lilium philadelphicum across Solem Township in 2021 and threatening the global supply of orange pin flags in the process, Jared came to his senses and focused on Echinacea in summer 2022… oh who are we kidding, Jared mapped a bunch of lilies in summer 2022. Rather than census flowering plants, Jared established seven 60 x 40 meter plots across sites with varying burn histories and mapped all flowering plants within those study plots (N = 361 flowering individuals). Pods were harvested from 23 plants.
Echinacea who?
Start year: 2021
Location: Remnant patches of prairie in and around Solem Township, MN
Overlaps with: Sleep
Data collected: Spatial and demographic data housed in the remlp Bitbucket repository
Samples or specimens collected: Pods/seed collected during summer 2022 currently reside in Jared’s office. These seeds need to be cleaned, counted, and scored for seed set.
Products: Stay tuned!
You can read more about the Lilium fire and flowering project, as well as links to prior flog entries about this experiment, on the background page for this experiment.
Prescribed burns increase the flowering rates of Echinacea angustifolia, but what aspect of fire induces flowering? Researchers have proposed many factors, including light, heat, nutrients, decreased competition, and smoke. Applications of liquid smoke increase germination rates in many plant species, but very few studies have tested the impacts of smoke on flowering. Our smoke experiment investigates whether liquid smoke will increase flowering rates of E. angustifolia. Many members of Team Echinacea have proposed this experiment in previous years, most recently Amy and Scott in 2019. However, this is the first year of installing the experiment in the field.
Applying the smoke treatments to a flowering Echinacea!
On July 29th, Alex and I visited the Hutchings property and recorded demographic data on 100 Echinacea plants that Scott and Amy had mapped in 2019. After further discussing methods and sample size with Jared and Stuart, we revisited the Hutching’s property to find additional plants. On September 20th and 22nd, Alex, Manogya, and I mapped and recorded demographic data for 205 more plants using the GPS.
We applied the first half of the smoke treatments on October 27th and 28th, and you can read more about that trip here. We applied liquid smoke to 110 plants, exactly half basal plants and half flowering plants for a balanced experiment. We used 11 different concentrations of smoke in our applications. We plan on conducting the second half of the experiment with an additional 110 plants in the spring.
Data collected: Methods, datasheets, and treatment groups can be found in Dropbox at ~/dropbox/burnRems/smokeExPt1. All smoke demographic data collected in summer of 2022 can be found in the aiisummer2022 repo at ~/aiisummer2022/smokeExpt/smokeExpt2022DemoData.csv. This includes coordinates, flowering status, rosette count, and head count for 305 plants. Demographic data will be collected on the plants once they flower in the summer of 2023. The stake file for smoke plants can be found in Dropbox at ~Dropbox\geospatialDataBackup2022\stakeFiles2022\stakeSmokePlants.csv
Samples or specimens collected: None at the moment
Products: None…yet!
Click here to read more about the smoke experiment!
During summer 2021, we began collecting data in remnant patches of prairie to quantify fire effects on the reproduction of Porcupine grass (Hesperostipa spartea). In summer 2022, we decided against collecting further data. A few very late spring burns that seemed to affect Hesperostipa, our smaller than expected summer crew, and a long list of projects led us to conclude it would be better to prioritize other projects.
Start year: 2021
Location: Remnant patches of prairie in and around Solem Township, MN
You can read more about the Fire and seedling fitness in remnants experiment, as well as links to prior flog entries about this experiment, on the background page for this experiment.
The goal of this research is to figure out a method and protocol to collect nectar from prairie plants. Once a collection method and protocol have been designed, the nectar can be analyzed for quality and quantity. We plan to compare prairie plants pre- and post-burn to investigate whether nutritional resources for pollinating insects differ before and after a burn. If there are differences, we can make recommendations about conducting burns and identify plants to use in restorations and reclamations that will provide the nutrients for pollinators.
A microcapillary tube inserted into an anther
Here’s a draft protocol and notes from summer activities….
Pre-collection:
Select heads ahead of time that will be flowering, preferably on day two of the male flowering stage, on the nectar collection day. Day two tends to have the most average anthers present on a head. Anthers tend to have more nectar than the styles, so the anthers will be our targeted area of collection. On the selected heads, place the crowns and pollinator exclusion bags over the heads. If the head has a twist tie for identification, use the same color twist tie on the bag.
Label the microfuge tube in ascending order prior to going into the field. Place a large binder clip on the last numbered tube and leave it in the supply so the next person to number knows where to start. Clearly number the tubes with a fine tip or medium tip permanent marker.
Nectar collection procedure:
Select the plants to sample from during the course of the day, I used seven.
If necessary, record the row, position, twist tie color, estimated male flowering day, and time of collection for the selected head to sample. If in a remnant, record the location using the GPS.
Prepare the microcap(s) and leave it in an empty microcap tube in the styrofoam holder in the field collection box so you are ready to collect the sample.
Remove the pollinator exclusion bag from the selected head.
On the same head, select the anthers that are the most recently presented to sample from.
Use a Q-tip to remove the pollen from the selected anther.
If the pollen is still in the way, use a Q-tip or end of a toothpick to move the anther out of the way.
Insert the microcap tube into the anther floret. Insert the microcap down into the floret until there is light resistance when you reach the base of the floret.
Twist or rotate the microcap five times. Remove the microcap.
Carefully remove the microcap.
Record the amount of nectar collected in the microcap in millimeters (mm). It is helpful to hold the microcap up to the sun to see the refraction of light from the nectar to see the amount collected. You can use the magnifying glasses to read the amount of nectar in mm on the ruler.
Place the entire microcap with the collected nectar sample into a numbered microfuge tube and record the number of the microfuge tube.
Securely close the top of the microfuge tube and place it into the cooler or pouch with the ice packs.
Replace the pollinator exclusion bag.
Label the flag marking the plant with the row and position for future reference and time efficiency. Once this task is completed, it will not need to be done for the same plants in the future.
Repeat steps 2-15 for the next plant.
Once the seventh head has been completed, head back to the first plant to repeat the entire collection procedure. This should take about 50-60 minutes to collect from 7 plants.
Upon returning to the Hjelm House, place the collected samples in the collected samples box, which then is stored in the refrigerator.
Field supply checklist:
Nitrile gloves
Microfuge tubes (tall enough to put the microcap inside and close the cap)
Microcaps
Drummond Scientific Microcap 1-000-0033 Microliter Pipets, 3.33µL Capacity (Pack of 100) 0.34 mm or 0.0133 inch diameter.
Two ten gallon buckets (one for sitting and one to set supplies on top)
Cooler with ice packs (ice packs can also be held in our pouch instead of the cooler)
Ruler with mm markings
Water and sunscreen
Clipboard with the data sheet
Rubber band to hold the data sheet onto the clipboard
Duct tape paint holder filled with paint and toothpick
Q-tip bundle
Wooden toothpick bundle
Field collection box to hold small supplies
Kestrel weather unit
Measuring tape or meter stick
Field notes:
This data was collected in experimental plot 1 from 7/18/2022 through 7/28/2022.
Sometimes the magnifying glasses directed sunlight to a focal point and it became quite warm (similar to killing an ant with a magnifying glass scenario).
I found it easiest to put the supplies inside the buckets while traveling to the site because it was a lot to carry.
I liked the magnifying glasses because I could adjust the lens up and down while going back and forth from the head to the supplies. I recommend wearing a bandana in between your head and the glasses for comfort, sweat, and keeping away long hair.
I recommended using the 3 uL capacity microcaps. They fit best in the floret and collected the most nectar.
I used a piece of Styrofoam to hold the microfuge tubes that contained the various sizes of microcaps, toothpick, pollen Q-tip, and the next empty numbered tube for the nectar.
Sometimes the microcap got bubbles in the tube, so I only counted the amount of liquid I saw. It seemed on occasion that some of the nectar went out of the tube, so it may be best to record the amount after sampling each floret. *A recommendation for next time: Place your finger over the end of the microcap to try and prevent liquid from being removed.
The pollinator exclusion bag being placed prior to collection increased the amount of nectar collected. Note, on 7/25, I sampled some heads that did not have pollinator exclusion bags prior to sampling and found that they had about the same amount of nectar as ones that had the bags at that same time of day. This will need further investigation. I also sampled from each of the most recently presented anthers that day to see if I would get more per head in one sampling rather than multiple times a day. I got more nectar per head, but I also sampled more anthers on this day. The data table below shows the average amount of nectar in mm per floret for each day of sampling. Note that the day I sampled the most florets in the afternoon, 7/25, has the least amount of nectar per floret. This day was also cloudy and some of the heads included in the data didn’t have pollinator exclusion bags prior to collection.
7/18
0.406 mm
7/20
0.399 mm
7/21
0.448 mm
7/25
0.213 mm
Pollinator Exclusion Bag vs. No Pollinator Exclusion Bag
As of 7/25, the time frame of 2:30 and 3:00 pm has had the highest average amount of nectar collected. The mornings and cloudy days have produced the least amount of nectar. After the first three days of collection, we noted that the mornings had less nectar, so I focused on mid mornings and the afternoons for the following days.
Time of day vs. mm of nectar collected
Start year: 2022
Location: Experimental plot 1
Overlaps with: NA
Data collected: Britney has a datasheet from summer 2022 that lists all samples and the conditions associated with their collection
Samples or specimens collected: All nectar samples were delivered to Dr. Rahul Roy at St. Catherine’s University on November 10th
Products: Stay tuned!
You can read more about the nectar experiment and find links to prior flog entries about this experiment here.
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. Currently 40% of net Minnesota State Lottery proceeds are dedicated to growing the Trust Fund and ensuring future benefits for Minnesota’s environment and natural resources.
Every year since 1996, Team Echinacea members record flowering phenology, taking measuring data and harvest heads of thousands of Echinacea angustifolia plants in common garden experiments. These experimental plots are prairie restorations and abandoned agriculture fields that are managed as grassland habitat. Some plots have multiple ongoing experiments within. Currently, the Echinacea Project currently has 10 established experimental plots.
This project status report will contain updates on experimental plots experimental plots 1, 2, 4, 5 and 8. Reports for the remaining experimental plots can be found on separate posts including Amy Dykstra’s plot (exPt03), the hybrid plots (exPt06, exPt07, exPt09), and the West Central Area common garden (exPt10).
Jo, Emma, Kennedy and Sophia measuring in the picturesque views of exPt02.
exPt01: Experimental plot 1 was first planted in 1996 (cleverly termed the 1996 cohort), and has been planted with nine other experiments in subsequent years, with the most recent planting being Amy Waananen’s inter-remnant crosses. It is the largest of the experimental plots, with over 10,000 planted positions; experiments in the plot include testing fitness differences between remnants (1996, 1997, 1999), quantifying effects of inbreeding (inb1, inb2), and assessing quantitative genetic variation (qgen1). There are also a number of smaller experiments in it, including fitness of Hesperostipa spartea, aphid addition and exclusion, and pollen addition and exclusion (the latter two experiments were continued the summer of 2022 and will have separate update posts). In 2022, we visited 7,273 of the original 10,673 positions planted and found 2,985 alive. Only 1,111 plants were classified as “flowering” in exPt01 this year. This is a drastic increase from the mere 79 plants that flowered in summer 2021– coincidentally, the plot was burned in the spring prior to summer 2022 and not prior to summer 2021. In summer 2022, we harvested 1,588 total Echinacea heads in exPt01. No additional staples were added to the experimental plot this year.
Some numbers for experiments within exPt01
Inb1: The INB1 experiment investigates the relationship between inbreeding level and fitness in Echinacea angustifolia. Each plant in experiment INB1 originates from one of three cross types, depending on the relatedness of the parents: between maternal half siblings; between plants from the same remnant, but not sharing a maternal or paternal parent; and between individuals from different remnants. All individuals were planted in 2001.We continued to measure fitness and flowering phenology in these plants. In 2022, of the original 557 plants in INB1, 92were still alive. Of the plants that were alive this year, 40 of them were flowering; this is a drastic difference from summer of 2021 where only one of the plants was flowering.
qgen: The qGen1 (quantitative genetics) experiment in p1 was designed to quantify the heritability of traits in Echinacea angustifolia. We are especially interested in Darwinian fitness. Could fitness be heritable? During the summer of 2002 we crossed plants from the 1996 & 1997 cohorts of exPt01. We harvested heads, dissected achenes, and germinated seeds over the winter. In the spring of 2003 we planted the resulting 4468 seedlings (this great number gave rise to this experiment’s nickname “big batch”). 1,467 plants in qGen1 were alive in 2022. Of those plants, 592 flowered this summer.
Other plots:
exPt02: To examine the role flowering phenology plays in the reproduction of Echinacea angustifolia, Jennifer Ison planted this plot in 2006 with 3,961 individuals selected for extreme (early or late) flowering timing, or phenology. Using this phenological data, we explore how flowering phenology influences reproductive fitness and estimate the heritability of flowering time in E. angustifolia. In the summer of 2022, we visited 1,856 positions of the 3,961 positions originally planted. We measured 1,438 living plants, of which 651 were flowering, with a total of 1,168 flowering heads. In the fall, we harvested 558 heads from exPt02. We began harvesting on August 10th and completed harvesting on September 12th. The large difference between the number of heads and the number harvested has to do with high levels of seed predation, mainly by ground squirrels. This year, Will, Jennifer, and other members of Team Echinacea published a paper in the American Journal of Botany using data from exPt02 – check it out here! Location: Hegg Lake WMA
exPt04: Experimental plot 4 was planted to gauge whether Echinacea from small remnant populations could be genetically rescued via an outcross to larger, more genetically diverse populations. Caroline Ridley and other members planted this plot in 2008. We did not visit exPt04 this year. Location: Hegg Lake WMA
exPt05: The only experimental plot planted at Staffanson Prairie Preserve (SPP), exPt05, was planted to compare progeny of maternal plants from burned and unburned sections of SPP. There were 2800 plants planted originally, but high mortality made it impractical to visit the plot row-by-row. Now, we and treat the plot like demography. We use our survey-grade GPS to find plants in exPt05 that have previously flowered and add more plants to the stake file if new plants in the plot flower. In 2022 we found 11 living plants, four of which were flowering! There were two heads that should’ve been harvested, but Alex and I forgot to harvest them (oops). Location: Staffanson Prairie Preserve
exPt08: Team Echinacea established quantitative genetics experiments to quantify additive genetic variance of fitness in Echinacea, with the idea that we can estimate evolutionary potential of study populations. The maternal parents of qGen2 and qGen3 are plants in the 1996, 1997, and 1999 cohorts. These plants were crossed with pollen from plants in remnants to produce seed for qGen2 and qGen3, which now inhabit exPt08. Originally, 12,813 seeds were sown in the common garden. Seeds from the same cross (shared maternal and paternal plants) were sown in meter-long segments between nails. A total of 3,253 seedlings were originally found, but only 363 plants were found alive in 2022. There were 14 flowering plants in 2022, and 15 heads. On a side note, 3 additional flowering plants were found in t-plot, but the heads were eaten before we could harvest any of them. Location: Wagenius property
Start year: Differs between experiment, see above. First ever experimental plot was in 1996.
Location: Differs between experiment, see above.
Overlaps with: …everything!
Data collected: Raw measuring data can be found in cgData repository. Processed data will be eventually uploaded to SQL database. Currently, SQL database has measuring data up until 2021.
Samples or specimens collected: See above for total harvested heads in each plot.
Products: Many publications and independent projects.
I want to study the impacts of isolation of Echinacea angustifolia on pollination. The remnant prairies, in which Echinacea is a model species, vary is size, and within the remnants the density and number of Echinacea plants are not consistent. I am hoping to learn if the size of the remnant or the proximity to neighboring plants has a greater impact on seed set, which is an indication of the degree of successful pollination.
Statement
In my spare time I like be outside with friends and family running, hiking, skiing, and biking. I love exploring new places in addition familiar ones.
Undeclared major (potentially biology or chemistry), Carleton College, Class of 2025
Research Interests
I would love to know more about any aspect of how the world works, because it is an interesting place. However, I am especially interested in conservation efforts in any field since the earth is the one planet we know of with life and biodiversity within that life, but changes to the ecosystems are causing problems. In the Echinacea project, I would like to know more about different characteristics of the plant that are changing through the years with various treatments.
Statement
I am from Roseville, Minnesota. In my spare time I like to run, ski, and ride horses, which are all recently acquired hobbies. I also like to read and bike.
This is me at a boardwalk with marsh, prarie, and forest habitats. This is me skiing at Carleton
