July 2009 Archives

Hello friends,

We just finished a rousing lunchtime discussion on the virtues of archiving, so I thought this might be an appropriate time to post our compiled data set from all four days of pollinator observations and captures.
compiledpollenobs2009.xls

Here are files with presence/absence of species within the 2m floral neighborhoods
fnc10mNeighborhood.csv and within 10m floral neighborhoods fnc2mNeighborhood.csv. The first is just a reorganization of FNC.csv and the second includes info from FNC.csv and from WITHIN10M.csv.

Here's a photo of the box I built for drying plants with generously donated materials from the Wagenius family.

A pressed specimen of Anemone canadensis collected at Hegg Lake with Greg.

One of my painted heads after being repainted this week. Each paint color identifies a pollen treatment.

Thanks to Mimi for letting me use her camera!

I am slowly moving up from the basement to rejoin the world at ground level. Thanks to Caroline for the labels - three sets of slides of co-flowering plants of E. ang. are complete. The pictures are waiting to appear online. Meanwhile, succession of the Hjelm house basement table (door) occurs. As a flowering plant replaces another on the prairie, Stipa spartea seeds have replaced the slides.
Can you see a difference between the pictures?
Wouldn't you like to peg some Stipa and populate the board?
I think Someone (codename: drone or riddler) may offer a 6-pack to the person who fills the last seed in each hole of each styrofoam "board". It would be more fun than chasing a chipmunk out of the Hjelm house.

Here is a list of the plants I used in the common garden for my experiment on pollen interference/competition. I used 20 randomly selected plants in the 96 garden with 12 pollination treatments on each plant. The heads I harvested are also included in this spreadsheet, but here they are again: 39 952 blu and wht heads were harvested on 7/24/09.

Baggins.CG.09.xls

The preliminary results seem to show that the only treatments that consistently did not shrivel were:
silver-the control, no pollinations
white-Carduus acanthoides pollen only (thistle)
pink-Coreopsis palmata pollen only

All of the treatments that received echinacea pollen (either am or pm) showed somewhat consistent shriveling.... will it hold up to the stats.... we will see!

the most interesting treatment is.....
purple-Heliopsis helianthoides pollen only- this one had a mix of results, which may be due to differences in the amount of pollen arriving on the styles or some other factor. but it certainly did not show consistent style persistence. hmmmmm.....

Just as a reminder, style shriveling indicates that compatible Echinacea pollen has arrived on a style, and can be a good predictor of seed set. Style persistence indicates that compatible pollen has not landed on a style. In the case of some of my treatments which had both Echinacea pollen and another type of pollen, shriveling may not indicate seed set. This will be tested by dissecting the heads and weighing the seeds later this year.

More results to come soon...

Listen up, Echinacea fans!

I've now finished making slides and taking photos of the first 68 insect visitors--only 107 left to go.

Here are some photos of the process:

1) Here is an insect carrying LOADS of pollen (haha! get it?) which I am about to transfer onto a small agar cube on a microscope slide.

2) I heat the slide, complete with pollen-covered agar cube and cover slip, on a hotplate. Next I throw the completed, labeled slide under the microscope camera and take photos like these:

3) I've pinned each specimen with a unique ID code that corresponds to its vial ID number.

The most common genera near as I can tell from the reference collection are...

Male Melissodes sp.

And Ceratina sp.

Please leave questions or comments and I'll do my best to respond!

-Amanda

On Medicago sativa in CG--Melissodes?

Beetle on Achillea millefolium

On Heliopsis helianthoides

On Monarda fistulosa

Some flowers aren't as easy to land on as Echinacea...

On daisy

Ant on Symphoriocarpos albus

thanks Gretel for letting me use your camera!

So, I've been working hard creating slides from all the styles everyone helped to collect (thanks!), and this is the protocol I've been using:

II. SLIDE PROTOCOL:
A. ORGANIZE THE STYLE VIALS BY SITE
B. CREATE SLIDE LABELS
C. RANDOMIZE THE STYLE INFORMATION ON EXCEL
D. CREATE THE SLIDES IN THE RANDOM ORDER GIVEN BY EXCEL
1. Place the blank slide on a clean surface
2. Pull out the correct vial, open carefully (sometimes the styles are on the lid of the vial), use clean tweezers to remove the contents. Place vial contents onto the slide.
3. Remove any anther parts from the slide; organize the stigmas so they are separate/easily differentiated from each other.
4. Place one drop of glycerin on top of the stigmas. If there are any bubbles, try to move them away from the stigmas.
5. Place a cover slip carefully over the glycerin and allow it to settle.
6. Use mounting medium to seal the cover slip to the slide. Allow the slide to dry on a flat surface for 24-48 hours.
E. Put completed slides into slide boxes.

Of the approximately 380 slides I started with, I have 150 left to do, so I'm more than 1/2 way there. Once I've created all these slides, I'll start taking photos and uploading pictures.

Anyway, just felt like it was about time I updated. Gonna go make slides now... ^_^

Here is the data that's been compiled for FNC, pollinator observations, within 10 m, and the isolation measure of flowering Echinacea focal plants.

Some things I wanted to point out that may or may not make a difference:
>In the FNC data, all quadrants with none present have no data for distances, and sometimes there is only one distance if there was only 1 infl of a species within a quadrant...
>I still need to check each tag number that we recorded for FNC and make sure it coincides with the original record of tags we made when we flagged.
>For the isolation measure, I put in 11 for distances >10m. I marked the original distance in the notes in cases where they were >10m but measured out (i.e.SPP)
>For inflorescence counts>100, I put in 1000. In the comparison file below, we changed 1000 to 101 since we had to sum the inflct for each unique ID and so some of the infl ct were showing up as >1000.
ISOLATION.csv
FNC.csv
POLLOBSWITHFLIES.csv
POLLOBSWITHOUTFLIES.csv
WITHIN10M.csv
pollcomparison ecan mesa amca.csv
ech mesa comparison with mean std error.csv
ech amca comparison with mean sterror.csv

As of 8/3:The last 2 files are new. In order to make the graph that appears on my poster, we divided the unique Id's into 4 groups: 1-alfalfa only in fl.neighborhood 2-ech only 3-both 4-neither. I took the average and standard error from each of those 4 groups to make the 4 bars on the graph. I want to do the same thing for Amorpha. So I attached the file that I'd use to make the same graph but with amorpha. You could use the third to last file which has infl ct for each unique ID for ecan, mesa, and amca to do the analysis but I figured I'd put the others up so you could know how I made the graph...

FYI, after Amanda and I looked at all the vials yesterday, it turns out that over our 4 day stint we had 132 bees, 13 scsf, 33 flies, 4 butterflies, 15 beetles. Good work everyone!

I attached a map of locations in the CG where we will plant Stipa seeds. Black dots are Echinacea locations, red circles are potential Stipa locations, and blue dots are locations we will plant this year. We will plant each Stipa seed relative to an Echinacea location, maybe 10 cm South or 20 cm North. Any thoughts?

mapOfStipaInCG.pdf

Greg and I have been collecting plants and pressing them. I am doing a plant collection with plants mainly from landfill for one of my classes next year at McGill in Plant Systematics. Greg is collecting specimens of anything that co-flowers with Echinacea and making slides with the pollen from each specimen to compare with the pollen found on the pollinators and styles from pollinator competition experiments. Greg, here is a site with information on collecting plants from the Missouri Botanic Gardens

Here is a document from my class that also describes field collection techniques:
HELPFUL HINTS2010.doc

I also made a drying box for the pressed plants, which is almost finished...

This post is long overdue! I thought I would give a little update on what I've been up to here at the Chicago Botanic Garden.

So to remind those who may have forgotten who I am and what my project is all about.. My name is Andrea. I attend McGill University where I study botany (with Allegra!) and over the past two years I've developed a very strong and deep DEEP love for life in the soil. Don't get me wrong, I love studying plants, but at this phase of my life soil organisms (namely fungi) dominate. It started with an Introduction to Fungi course I took two years ago that involved a series of guided and independent mushroom forays and culminated in the production of a mycological portfolio. I then took a biology of fungi course that was much more thorough and only confirmed for me just how awesome fungi really are. Last summer I was lucky enough to have the chance to work as an REU intern with Louise Egerton-Warburton, soil scientist at the Garden. My research looked at how microbial communities respond to buckthorn invasion and subsequent restoration. I'm back this summer to continue research on that project and as well to start looking at the mycorrhizal communities associated with the Echinacea populations in the remnants of interest in Minnesota.

The main goal of my experiment is to see if there is a relationship between plant genotype and mycorrhizal community. At the beginning of the summer Stuart helped me select nine families in the '97 common garden and from each family around 9 individual plants were randomly chosen.To collect the mycorrhizal fungi I use mesh baggies that have two polycarbonate membranes secured at the bottom. One of these membranes I will stain and permanently fix onto slides for the purposes of determining hyphal length. This will give me an overall idea of mycorrhizal density and colonization. Hopefully I will have time to extract DNA from the other membrane to get a closer look at who's really there.

To install the bags, I basically wedge the bag carefully into the ground near the roots of select plants so that the membranes sit at least 4 cm below the surface. Over time, the mycorrhizal fungi growing near the roots will run across these membranes and have no trouble meandering through its pores. Little do they know that it's a trap!! Around each plant I placed one bag and waited about three weeks. Recently, I retrieved those bags and I've spent the past few days making slides. I promised to post pictures so here are a couple exhibiting some spores and hyphae!

(Image taken with 40X objective)

For some reason it won't let me load more than one photo so I will try again later.

More updates coming soon!

-Andrea

This week we are going to make big dents in CG measurements and our independent projects. We will measure the CG MWF morning and TTH afternoon. We'll start T & Th morning with Phenology assessments and end off each morning with time to work on independent projects. Afternoons on MWF will be devoted to independent projects.

Be ready 8:30 Monday morning--with sunscreen on & visors synced--for a pep talk (it'll be about efficiency & the well-oiled machine)!

day        AM                PM
Monday     measure CG        ind. proj.
Tuesday    phenology & i.p.  measure CG
Wednesday  measure CG        ind. proj.
Thursday   phenology & i.p.  measure CG
Friday     measure CG        ind. proj.
Saturday   phenology & off   off

I was glad to participate in assessing floral phenology Wed morning and, with Amy, checking to resolve uncertainties remaining from this year's monitoring of the first recruitment experiment (not to mention a very fun lunch with the team!). We sampled tissue from closely neighboring rosettes, where it isn't clear whether they are the same or different plants, for eventual molecular analysis in Chicago by Jennifer and her team. Resolution of those plant identities should certainly help reduce the problem of counts of survivors *increasing* between censuses. But, in retrospect, I wondered whether the info we recorded was crystal-clear in terms of how this year's counts should be adjusted, depending on the outcome of the IDing, particularly for the zones where many seedlings were recorded. When the remaining double-checking is done, it would be good to keep this in mind...

Of the many, many other terrific things that I'm excited are being accomplished, I'll just comment that I'm happy to see Megan's post that she has sampled pollen and stored it in different conditions to check its long-term viability. Finding a way to keep pollen viable for a month to a year would pave the way for experiments I thought up while observing pollinators out at LF on July 7. I see that Megan noted the amount of pollen available for that sample wasn't large, so it would be great if another set of samples could be taken, also so other plants are represented.

This two-part entry includes one observation about pollination that struck me odd. I had a floral head (A) at Loeffler's corner and as Agopostamas texanus approached - it stopped - flew backwards and away - and visited others nearby (all Echinacea). Did the presence of ants on the head - around the anthers have anything to do with the "I'll just come back later" actions of the bee? Has anyone else observed a head NOT get visited even though it was ripe with pollen because of the presence of ants?
My second half is simply noting that a calico cat and two large kittens were at the end of the common garden yesterday as I left about 4PM. The mother slunk away and the gold/white kitten watched me while the other kitten mostly white/ some black was trying to consume a chipmunk! Are these cats known to inhabit the area?

Gretel- KJ's
Mimi- Railroad Crossing
Allegra- Landfill East
Stuart- Staffanson
Amy- Steven's Approach
Amanda- Yellow Orchid Hill
Daniel- Aanenson
Kate- Riley
Caroline- On 27
Greg- Loeffler's Corner

See you guys at 7:30ish at Hjelm House!
xoxo,
Amanda

We have made a few changes to the protocol for tomorrow's FINAL outing for pollinator observation and collection:
1) DO NOT capture syrphid flies!
2) If you observe a small common syrphid fly (Family: Syrphidae, Sphaerophoria sp.) on your Echinacea head, make a note that one SCSF was observed. Here are some photos of syrphid flies:

http://echinacea.umn.edu/insects/images/poll2005vin1948side.jpg
http://echinacea.umn.edu/insects/images/poll2005vin1927top.jpg
http://echinacea.umn.edu/insects/images/poll2005vin1927front.jpg

Here's a video that might help:
http://www.youtube.com/watch?v=UkjPfAMGmiw&NR=1

---------------------------------------------------------------------------------------------

As for pollinator collection and analysis...

We've gone out collecting three times now (tomorrow is the fourth and final day), and we have about 140 Echinacea insect visitors in vials in the freezer. Here is my protocol for making slides and labeling specimens:
1) Remove vial from the freezer and allow insect to thaw
2) Cut a very small (2mm cubed) piece of agar and situate it in the middle of a slide.
3) Remove the insect from the tube and dab and sweep every surface of the body across the agar cube. This is intended to simulate the amount of pollen that might be transferred to Echinacea styles during a visit.
4) Set the insect aside on Styrofoam.
5) Sweep the inside of the vial with the agar cube for any loose pollen.
6) Place a cover slip atop the agar cube and put the slide on the edge of a hotplate on LOW heat. Watch the slide carefully and remove it when the agar is soft but not melted (once melting starts, it will quickly boil and create bubbles in the slide).
7) Pin the insect (for identification if it is an unknown species, or quick-n-casual for a known species) and pin it with a new one letter, three number code.
8) Label the slide with the vial code and the specimen code.

Photos:
I photograph the slides on the same day as I make them so that the agar doesn't dry out before the photo. To sample the slides, I have generated random pairs of numbers from 1 to 22 (inclusive). The cover slips are 22 x 22 mm and I use the ruler on the microscope to scroll to the randomly selected "plots" from one corner of the cover slip. From there, I take the photo at 40x. I take ten photos for each slide, and sometimes take more than one photo per plot if I capturing all of the pollen grains requires multiple focuses.

I will post some photos of this process soon, so hang in there!
As always, feel free to leave questions or comments.

Thanks!!
Amanda

So, there has been a serious lack of pictures lately (aside from those awesome Stipa scans), so I am posting a bunch of pictures taken a while ago, just so you can see what we are up to:

http://www.flickr.com/photos/danrath/3727546804/
Amanda in her little corner of the farmhouse, doing voodoo with bees.

http://www.flickr.com/photos/danrath/3726738091/
http://www.flickr.com/photos/danrath/3726739525/
http://www.flickr.com/photos/danrath/3726736903/
Greg and Kate locked away in the Basement of Oppression

http://www.flickr.com/photos/danrath/3727538558/
http://www.flickr.com/photos/danrath/3727532876/
http://www.flickr.com/photos/danrath/3726727059/
http://www.flickr.com/photos/danrath/3727301314/
Berry Picking, starting with Caroline's Gollum face. We have picked at least 20 buckets of berries from these people.

http://www.flickr.com/photos/danrath/3727297280/
http://www.flickr.com/photos/danrath/3726488747/
http://www.flickr.com/photos/danrath/3726486389/ - Aphids, the enemy.
Insects in the Common Garden that I found while searching for plants

http://www.flickr.com/photos/danrath/3726483267/
Mimi broadcasting Stipa grass

We spent most of yesterday collecting pollinators and measuring plants in the Common Garden. (It has never taken me 3 hours to go 30 metres before, but all the plants I measured seemed to be in the middle of a grass clump). I have also figured out a procedure for refinding plants in the transects, and it should not take very long to refind all of them. This is good, as I might be working by myself to do that.

On July 20th I collected pollen,separated into three microfuge tubes, from plant 36, 958. Tube #1 has been left at room temperature, #2 is in the refrigerator, and #3 is in the freezer. There wasn't much pollen available, so I hope it's enough to try some pollinations and see how long the pollen stays viable under the three treatments.

Apparently there is some question about what I have been doing lately. Daniel was partly right--data entry has been on my list. Remember checking the recruitment zones earlier this summer? And being rewarded with all those six packs? Well, there were some ambiguities in the data. I compiled a list 2009 zones to check.xls of all the things we need to check in the recruitment zones. Some of these (checking burn status) can be taken care of with a visit to the plot; other ambiguities may be cleared up by taking some tissue samples and sending them to Chicago for microsatellite analysis.

Below is one of the scans of Stipa seeds collected from Douglas County. This particular set of seeds was collected from a plant at Staffenson Prairie Preserve. The seeds themselves are pointed towards the left side of the image, and extending from them are the long awns that give Stipa it's common name (porcupine grass). At this stage, they look less like quills, though, because they have dried and started to coil (click on the picture to see it full resolution and you can actually observe the coils and lots of other neat features of the seed, like hair and a dagger tip!). Out in nature, the coiling action would allow the seeds to attach to a disperser or to drill themselves into the ground in preparation for overwintering and germinating the next spring. We'll be scanning all of the seeds we collected (an estimated 3,000+ seeds from 431 plants), making digital measurements of seed length and width, and planting around 2,500 of them interspersed with Echinacea in the common garden.

Hi, Team Echinacea --

Its Diedre and Jake with an update from the lab at the Chicago Botanic Garden.

In spite of several setbacks, including a crowded lab and a power failure that shut the lab down for an entire day, we've been able to create a lot of data. Currently, we have ten microsatellite primers working which we use for paternity and genetic diversity analysis.

Recently we've been able to up our extractions to over 150 samples a week and 10 PCR's a day! Jake and I started extracting the samples that Jennifer and I took in Minnesota several weeks ago. Jake is using these for his poster on population structure. He will be looking at whether there is interbreeding or inbreeding among the nine remenant populations we sampled (East of Riley, Anenson, Steven's Approach, Landfill, Railroad Crossing, Staphenson Prairie Preserve, KJ, and Ness). Jake has already found some interesting results with the use of Structure and FStat.

The poster I am working on will look at the diversity of pollen donors with regard to flowering on individual and population levels.

Here are some pictures of our work in the lab:

Jake and I making DNA extractions a little more fun than they already are through use of our artistic talents.

Some preliminary results for Jake's project.

We are going to plant Stipa spartea seeds into the common garden. Here is a
file with target locations for ~2600 seeds. The seeds will be planting in ~269 batches. Here's a list of the batches: allStipaStarts.csv Sometime before we plant, Caroline will share a photo of one of those Stipa seeds.

Here's some of the work I've done with organizing my data. I still need to figure out how to organize it to be able to analyze it, so this is mostly just preliminary work. I have about 2 weeks to put this all together....any help/advice is appreciated because right now, the data I have is a little overwhelming. There are 3 sheets in this document.
Ech Guide to Co-Fl Sp.xls

For next week, it looks like the weather should hold up for Tues and Thurs to be able to do pollinator observations. So we will need to flag the sites on Monday and have everything ready to go for Tuesday. Remember, you ALWAYS record something for each observation you make, regardless of whether or not you observed/caught a pollinator. Select No for poll. observed and No for pollinator caught if this is the case. Some things I wanted to clear up for people helping with FNC:
>If you reach 100 when counting inflorescences, stop and record >100.
>When recording the species within 10m, you will no longer put this into a memo. Instead you will always select pl A, record 0 for infl ct, and in the field of quadrants, select the fifth option called "within 10m".
>Review the guide to co-flowering sp for how to count infl or print one up and ask me if you have questions.
>If you come across a new species that isn't in the list of species in the form, record in the notes not only the species but also a brief description of how you counted inflorescences.
Thanks!

Here's some of the pollinators I saw on Coreopsis near Hegg Lake. They seemed to only be pollinating Coreopsis although there were other species like Achillea, Amorpha, and Echinacea around.

On Friday all of us except Greg went on a trip to a mesic prairie 3 hours away to help Gretel look for orchids. We split into 3 groups of 3 and flagged the flowering plants within the various treatment grids.

The Western Fringed Prairie Orchid, Pratanthera praeclara a threatened species. The first orchid I've seen in the wild!

Allegra, Amanda, Daniel, Caroline, Amy, Stuart--it was pretty cold for mid-July!

Mountain mint--Pycnanthemum sp. It was really neat to see some different species found in the mesic prairie, as well as some familiar ones. Some others plants we saw were Liatris, Rudbeckia hirta, Apocynum, Lilium philadelphicum,, Asclepias incarnata, A. speciosa, and Campanula. Below is another mint, whose name I can't remember:

Thanks Gretel for letting us help!

Hello all!

This is Daniel with another update on what team Echinacea has been up to in the past week. Allegra, Stuart and I have become what I like to call the "Staffanson Crew", and we are responsible for doing phenology at Staffanson every other day. Today we got the time down to about 2 hours and 20 minutes, from 3.5 hours originally. The process was made a lot more efficient by splitting up sections and giving everyone a separate checklist. Of course, the temperature was almost 40 below, so that was slightly unpleasant. Add the wind and the fact that I was only wearing 2 thin layers, and you have a recipe for hypothermia. However, I persevered, thinking of my avocado and sausage sandwiches waiting for me at the Hjelm house.

The pollinator project has been going along well, with Kate, Amanda and Mimi hard at working sorting out the oodles of data they have obtained. Greg and Kate are based in what I like to call the "Basement of Oppression", working on making slides and taking pollen photos. Amanda is pinning bees and creating agar slides with the different pollen loads, then photographing them. Finally, Mimi is working on sorting out all the different types of flowering plants found at each sites. Meanwhile, who knows what Amy and Caroline are up to? Reviewing papers and entering data most likely, tasks far beyond the comprehension of we undergrads.

In my case, I have been searching for the different plants in the common garden that we identified as having spittle. I spent all afternoon in the common garden yesterday, and it was a ton of fun, especially since I saw so many interesting things. The most interesting thing of all though, was watching a bunch of ants pick up and move an aphid that was sitting on a leaf. The aphid may have been dead or alive (alive would be so cool!), but since I was silly enough to forget my camera, I guess I'll never know.

Most of the plants I looked at have aphids on them, but I will need to wait until I finish looking at them all before I draw any conclusions. Meanwhile, our transect searches are done until next week. However, I have found aphids on many of the plants I saw during our Staffanson searches, so I remain hopeful!

To celebrate peak flowering in the common garden, Megan made these awesome cupcakes. We all enjoyed them. Thanks, Megan!!

The common garden hit peak flowering status on July 13, 2009. The graph below shows how many heads have started and ended flowering each day. Approx. 197 heads have yet to shed pollen of the 1,614 found.

Please review this protocol for measuring plants in the common garden.

Kate and I made a list of species coflowering with Echinacea at Ri, LC, and rrx yesterday. These species are not within the 2m floral neighborhood, but are within 10 m of at least one of the observed plants. coflsp13jul2009.xls

Here's a snippet of code I used to generate files to upload to visors.

makeRandFileForVisor <- function(size = 50, fname = "xyz"){
write.table(sample(1:size),
                 file = paste("E:\\shared\\rand",
                                    size, 
                                    fname,
                                    ".txt", 
                                    sep=""), 
                 quote= FALSE,
                 row.names= FALSE, 
                 col.names= paste("rand",size,fname, sep="")) }
visors <- c("ag","dr","kg","ad","cr","gk",
            "mmj","mj","ah","gd","sw","rs")
for (i in visors) {
makeRandFileForVisor(20,i)
makeRandFileForVisor(50,i)
makeRandFileForVisor(100,i)
makeRandFileForVisor(200,i)
}

Ech jenkins FNC protocol revised.doc
Ech Guide to Co-Fl Sp.xls

I have a photo guide but I can't upload it b/c the file is too large.

If you plan on helping with FNC, please read the documents above. It's important that everyone counts inflorescences the same way. Thanks!

Also, for tomorrow, there are a couple of notes I thought I'd add:
>Please make a note if you see ants on the head of the plant you are observing.
>Also make a note if it is mostly cloudy.
>Try to get to your site with about 10 min to spare so you can get your supplies ready and orient yourself with the placement of the flags to avoid time spent wandering in search of flags.
>Please try to start your observation as close to 8am as possible. End at 11am. Do not start an observation if you can't finish by 11.
>Remember that you will only be collecting styles at the end of the observation pd from now on. Clean your tweezers with your shirt in between collections.

The little map on the right shows the locations of recent visitors to the Echinacea flog (since 29 June 2009). I just noticed that Hong Kong caught up to Belize (2 visitors each). These countries are tied for third (behind USA and Australia). Which country is going to have the most visitors by the end of the summer?

Here are the standings today (12 July)...

United States	182
Australia (AU)	3
China (CN)	2
France (FR)	2
Hong Kong (HK)	2
Belize (BZ)	2
Canada (CA)	1
Romania (RO)	1
Norway (NO)	1
Austria (AT)	1
Israel (IL)	1
Ecuador (EC)	1
India (IN)	1
Afghanistan	1
Japan (JP)	1

You can view current tallies to see where you favorite country ranks. Note that Ecuador is in 4th place and Hungary isn't yet on the list.

I did some calculations of compatibility rates between pairs of half- and full-sibs while working on a revision of a paper about inbreeding depression in Echinacea. I started on the back on an envelope, but quickly moved to the flog because I did the calculations before, but lost the envelope. Here are the basic questions:

What proportion of matings between full-sib and half-sib Echinacea plants is compatible? Calculating a simple answer assuming no dominance in S-alleles is straightforward, but unrealistic. I think I started in the right direction toward a better answer. Comments appreciated.

Thank you thank you thank you to everyone for helping this week. Just in terms of my project, we characterized floral neighborhoods for almost 70 plants in three days. In terms of pollinator observations, Tuesday's escapades in the remnants were fruitful, but thursday's weather would not hold out for us, so we had to postpone the second day of observations to next week, meaning we will have to randomly select a different set of 8 plants for all 10 remnants. I expect to see some more diversity in the neighborhoods next week because some species are just starting to flower like Coreopsis, Dalia, Apocynum, and Amorpha.
Here;s Amanda measuring to the nearest flowering Echinacea in Aanenson's:

Here's some pictures of our fun 4th of July and the amazing sustainable sandcastle.

Waniel, Per, and Hattie

Yesterday Daniel told us we could have a romantic walk in Staffanson Prairie if we came with him, but instead he made kate and I search his and Amy's transects. What a trickster. Here he is cursing the heavens.

A flock of pelicans flew overhead at NWLF.

I have many more pictures http://picasaweb.google.com/mimijenkins/MinnesotaSummer09# in case you're interested.

Hey everyone,

Thanks again for collecting helping with style collection, so far I've made four slides and the results are certainly interesting. There is certainly more pollen from 8am to 11am, but it's going to be hard to tell what pollen is there. So far, I haven't seen any thistle pollen, which is purplish, or any football shaped pollen, only a Echinacea and/or "-opsis" pollen (Heliopsis or Coriopsis). But then, my current sample size is tiny, so this may change. I have a lot more slides to produce, but so far so good!

FOR NEXT WEEK:

The protocol for style collection next week is a bit different, so please read this and take any notes that you feel are necessary, or ask me any questions you might have.

We will only be collecting styles at the end of the observation period. ONLY COLLECT STYLES ONCE!

Prior to taking the style off the plant, we will be recording style persistence data, to recap: The form will have five new lines for you to fill out, fr1, fr2, mr1, mr2, and immatures.

Fr1 will be the lower row of unshriveled styles. Count a row if there are more that 3/4 of the styles left.

Fr2 will be the upper row (ie the most recent) row of unshriveled styles.

Mr1 will be the first row of male anthers

Mr2 will be the last row of male anthers (usually there will not be a mr2, so leave this blank)

Immatures is the number of rows or florets left (put an "r" or an "f" to indicate which). Only count florets if there are 11 or less. Only do this if head is at the end of flowering.

One last thing, if there is still a problem with recording the letter field, enter the tag number of the plant in that space and enter the flag letter in the notes (if there is no tag, put a zero). There shouldn't be a problem, but if there is, just try to capture the information requested in the notes section.

Thanks again for your help and your patience!

-Kate Monster

This may (tentatively) be the official web presence of the pollen library - I am fairly comfortable using wikispaces and the students in my classes are as well. See if it works and provide any feedback you may have.

pollen.wikispaces.com

Enjoy the weekend.

For those who may find it useful soon. The ppt file (which is large) contains the partial identification key for our usage.
If you look closely, the diameters of the main four (Amanda has found) at this time are marked.
More to come next week.Who's who.ppt

Where is our study area? We focus on >6400 ha (25 square miles) of land that used to be tallgrass prairie and is now mostly used for agriculture (especially corn & soybeans). There are lakes and sloughs too.

The study area comprises these 25 sections:

T128 N R40 W:
31, 32, 33, 34, 35
T127 N R40 W:
6, 5, 4, 3, 2,
7, 8, 9, 10, 11,
18, 17, 16, 15, 14,
19, 20, 21, 22, 23

Plus, the area extends into the surrounding sections:
T128 N R40 W: 30, 29, 28, 27, 26, 25, 36,
T127 N R40 W: 1, 12, 13, 24, 25, 26, 27, 28, 29, 30,
T127 N R41 W: 25, 24, 13, 12, 1,
T128 N R41 W: 36, 25

Two files showing the design of the Echinacea-mycorrhizae experiment and the locations of the sampled plants:
designMycorrhizae2009.xls
dataSheetMycorrhizae2009.xls

This morning, Amy and I searched transects in Staffanson. We did random points 15, 36, 19, 5, 20, 40, 12, 28, 1, 14, 3, 37 and 27. They were 10 metre long and half meter wide transects, some of them half a kilometre apart. We found 1 plant (!) total in the transects, and 3 plants nearby.

*whew*. Lotta walking...

The Protocol for Style collection tomorrow is a bit different, so, everyone needs to get to the farm at 7:15 so we can go over it! But let me give you a quick overview of the main points:

• We will only be collecting styles at the end of the observation period. ONLY COLLECT STYLES ONCE!
• Prior to taking the style off the plant, we will be recording style persistence data:
o The form will have five new lines for you to fill out: fr1, fr2, mr1, mr2, and immatures:
ß Fr1 will be the first row of unshriveled styles. Count a row if there are any left at all.
ß Fr2 will be the last row (ie the most recent) row of unshriveled styles.
ß Mr1 will be the first row of male anthers
ß Mr2 will be the last row of male anthers (usually mr1 and mr2 are the same thing, ie the same row so just fill in the same number twice if this is the case)
ß Immatures is the number of rows with immature florets left, or if the number is less than 11 total (for the whole head), put that number down.
We'll go over this quickly tomorrow so everyone can get the idea.
• One last thing, if there is still a problem with recording the letter field, enter the tag number of the plant in that space and enter the flag letter in the notes (if there is no tag, put a zero). There shouldn't be a problem, but if there is, just try to capture the information requested in the notes section.

Thanks again for your help and your patience!

-Kate Monster

Two files: pollCompMemo06jul.txt and mimimemo070809.doc

Here's the file.

RESULTS FOR NAME THAT POLLEN

Slide 1: Heliopsis

Slide 2: Coreopsis

But don't they all kind of look the same??

Yesterday, between ten people at ten remnants, we collected...
68 pollinators!!
(That's almost 70!)

A big thanks to all who participated. You had an impressive capture rate and recorded your vials flawlessly.

We will go out to collect again tomorrow morning, each person to a different randomly chosen site (I will post these on the flog later today). Please arrive at Hjelm House at 7:30 to synch visors and pick up supplies so that we can all begin observations at 8:00 AM. I will provide muffins and coffee.

Here are some updates to the pollinator collection protocol-- please read them and jot them down (if necessary) on your printed protocol before going out tomorrow morning.

1) As you all noticed, the visor option for "pollinator observed" does not allow you to move on unless you provide a response. There are two methods for selecting either "yes" or "no". What I would prefer that you do is click on the words "pollinator observed" and click either "yes" or "no" on the resulting screen. The other possible method is to check the box for "yes" or check and un-check the box for "no", but this should only serve as a backup plan.

2) When entering your stopwatch time, please use a decimal between minutes and seconds. So, if it takes six minutes and twenty seconds for a pollinator to arrive, your entry should read "6.20".

3) If you observe but do not capture a pollinator, enter the data for the observation, select "no" for "pollinator captured?" and move on to the next plant. Do not stay at that plant to wait for more pollinators.

If you have any other questions or helpful tips for tomorrow's collection crew please write them in the comments and we will address them before tomorrow morning.

Thanks again, guys-- I guess dreams really do come true.

You know you could barely contain yourself with all the excitement and anticipation ... here are the handles revealed!

Allegra- Legos
Amanda- Robo Cop
Amy- Joker
Caroline- Riddler
Daniel- Yea Mon
Greg- GT
Gretel- Queen Bee
Kate- Monster
Mimi- Penguin
Stuart- Drone

So, for those of you who were wondering what Team Echinacea will be doing tomorrow, here is the field protocol for the transect searches that we will be using.

Field Protocol.doc

Any questions, please let us know!

Also, we were searching for Stipa today (a prairie grass that Dr. Ridley is planning to add to the common garden), and this is the setup we used to mark the sites with the GPS:

The antenna allowed us to get about a 9 cm margin of error when using the Trimble. And yes, that is yours truly manning the antenna, ensuring that the carrier lock is not lost. We were all ready to tell the next person who asked us what it was that we were searching for nuclear waste.

I have returned to take images of pollen as seen below. It will still be a few hours/days/more? to get the images as desired but this is a start. I am predicting some trouble to distinguish between coreopsis, helianthus, and echinacea so be ready to be distinguishing.
Attached are a protocol for pollen slides and the image of echinacea 7.1.09.ech7.1

Protocol for Pollen Slide Prep.doc

This file lists flags in random orders suitable for pollinator observation tomorrow.

Here's the R code used:

flagOrder <- function() {
cat(cat(sample(LETTERS[1:8]),"\n"),
    cat(sample(LETTERS[1:8]),"\n"),
    cat(sample(LETTERS[1:8]),"\n"),
    cat(sample(LETTERS[1:8]),"\n"),
    cat(sample(LETTERS[1:8]),"\n"),
    "\n")
}
for (i in 1:20) flagOrder()

Amanda, Kate, and I have combined our protocols and I've included a revised equipment list.
Echinacea PONS equip list.doc
Ech combined PollComp Protocols.doc

The following people have agreed (I think) to help with this project on the mornings of July 7th and July 9th, as well as July 21st and July 23rd. If you cannot for some reason, let us know tomorrow.

Stuart, Gretel, Caroline, Amy, Daniel, Greg, Megan, Mimi, Amanda, Kate. Thanks everyone!

Also, after some discussion with Allegra about her pollination and painting needs for this week, I think that we need to either A) give up one person and therefore one remnant to help Allegra or B) have the normal 4 observations for remnants with less than 8 plants instead of doing more observations within the 3 hr pd, and have those people help Allegra when they are done early with pollinator observations/style collection or C) a combination of A&B. Thoughts?

Note: I have included the protocol in with pollinator observations and style collections because we are using the same plants for all 3. However, 3 people can do this over the course of 3 pm's. Those who are helping so far are: Kate, Allegra (when not doing pm pollinations)

Friday we practiced catching insects for a half hour but much more practice is needed and those who were not there on friday need to be trained. I think a group training/practicing session before lift-off is imperative.

Today Daniel and Allegra helped me assess the flowering plants situation at some of the remnants we plan on using. We flagged flowering plants at: Riley (5),YOH (3), NRRX (4), LC (8), Steven's appch (3). Some of these sites had plants that seemed likely to flower by Tuesday so we flagged those plants as well.We randomly selected 8 plants for LC since there were 17 that were either flowering or seemed very likely to flower by Tuesday. There was only 1 plant flowering at NWLF so that site has been eliminated. We also noted co-flowering species for each remnant. Tomorrow we need to finish flagging and randomly selecting plants at the 5 other remnants.

Stay tuned for July 4th picnic and kick-ass sandcastle pictures!

Stuart, here are the estimates of people hours I will need for my project. Also, a schedule of times when I would need helpers during this week. Who wants to trade some hours?

Monday I would ideally like to paint and count styles for up to 80 treatments which I estimate will take 9-10 people hours (depending on how many need to be trained). I would like to do it after phenology or at least do the style persistence before lunch, painting can happen later. Wednesday and Thursday also follow the same schedule.

But... this would mean a big pollination day on Tuesday, when everyone is out in the remnants. Pollination takes about the same amount of time. So Mimi and I have come up with several possible solutions (or a combination):
- If Ruth will be here Tues. morning she could help collecting insects and someone else could help pollinate
- Cut the remnants down to 9 instead of 10
- Have some people in smaller remnants leave a little earlier than 11am, or have vehicles and be able to book it back at 11 to help pollinate from 11-12
- Cut down the treatments painted on Monday, however I don't want to cut down too much, since some of my plants have been flowering since Thursday or Friday and I don't want to wait too long
- Skip painting on Monday, paint Tuesday and Thursday, and pollinate Wed. and Friday when we have more people to spare

- I would also like help collecting pollen on Tues. and Wed. but I may be able to do this myself if needed.
- lastly, I need one person to help me do afternoon pollination on Tuesday and Thursday between 2-4pm

Sorry for throwing this into the mix so late in the game folks. I guess it was just hard to know how many hours and people I would need before we did a full day of painting and pollinating.

-Allegra

Carex molesta


Carex gravida

Lythrum alatum


Sporobolus neglectus

Here are some more pics of species I need for my project. Thanks for everyone's help so far.
Agalinis tenuifolia

Teucrium canadense

Your choices are: Heliopsis, Coreopsis, and Echinacea. (hint: there is only one species of pollen in each photo.)

Put your guesses in the comments, and we'll reveal the answers on America's birthday! Go ahead, put some money on it.

-Daniel and Amanda

EDIT-- We will not be announcing the results until there are at least three guesses in the comments!! Get guessin, folks. And happy July 4th!

Today we got the microscope camera in the mail-- here are the results!

This is the long-awaited photo of Echinacea angustifolia pollen. THIS IS IT, GUYS. Are you crying yet?

Love,
Daniel and Amanda

Over the years I have made several notes about locations of Asclepias viridiflora individuals. I have not noted the species at Staffanson Prairie Preserve. I've copied notes below. I can show you where these plants are (on a map or live)...

2-July-1998 site eth
Asclepias vividiflora 6.5 paces S of 2294

1-Aug-1998 site eth
Asclepias viridiflora w pod!

23-July-1998 site nolf
EA pla #3069 cf Asclepias viridiflora 1.1m WSW of this EA

I have mapped an Asclepias viridiflora individual at NRRX. No notes, just the location.

I have collected several seed pods from A. viridiflora at the landfill. Here are the records...
Landfill 9/5/1997 26 seeds 1 pod 4 planted at TP plot
Landfill 9/5/1998 3 pods

Finally, here's a note from my visor from earlier today. The yellow flags are at your prairie turnip plants.

Note-to-megan
7/3/09 9:31 am
landfill
Asclepias viridiflora
2 fl plas between
yel flags 1-02 & 1-28
1 fl pla between
yel flags 1-31 & 1-52
1 fl pla SSE of
yel flag 1-47 (far S) in dip

It might be fun to see Prairie Home Companion-- Free!--in Avon Minnesota.

For some time, the powers that be on the Echinacea project have thought, "Wouldn't it be nice to compare all we've learned from coneflower with another important native component of the prairie?" Well, this summer, study of habitat fragmentation and its ecological and evolutionary impacts broadens to include an additional species. And the winner is ... Stipa spartea aka porcupine grass! The time is right now to collect the seed, which will be sown into the common garden later this summer.

Here's a link to a page from the Bell Museum of Natural History about Stipa spartea.

Below, you will find the sampling protocol that Greg and I used to collect seeds from Staffenson Prairie this afternoon. Stuart may have thoughts to add and the strategy might change a bit as we run into the reality of different Stipa remnant populations. In addition to remnants, we will also be collecting seeds from roadside transects. Expect that challenge to be met later next week.

Protocol for sampling Stipa spartea in prairie remnants.doc

Hello everyone,
I'm glad you are all going to visit, neighbors. The echinacea is about to flower in full force, as are the leadplant (not today but soon), coreopsis in abundance, some goldenrod, purple prairie clover, dalea. Additionally galium, phlox, alumroot, ground cherries and lilies are blooming.
There will be many pollinator neighbors out next week trying to make a speedy delivery. If the weather is nice Mon AM, why don't you join us, neighbor?
See you all Mon.!

As promised, you can read the protocol I've developed for collecting pollen styles from Echinacea plants. This protocol will be joined with Mimi and Amanda's to form one complete and very awesome experiment. You can find my protocol here:
Kate's Proposal_1.doc

Input welcome!

-Kate

Cirsium altissimum

Asclepias viridiflora

Elymus virginicus

I generated a list of 40 random UTM coordinates for SPP and posted them here: sppRandCoords.csv.

Here's the R code I used to generate random coordinates...

df <-  data.frame(order= 1:40,
                  E= round(runif(40,  286100,  286900),2),
                  N= round(runif(40, 5077080, 5077500),2))
write.csv(df, file= "sppRandCoords.csv", row.names= FALSE) 

I gleaned the rough SPP corner coordinates from Google Earth--UTM 15T:
NE 286900 E 5077500 N
SE 286900 E 5077080 N
NW 286100 E 5077500 N
SW 286100 E 5077080 N

Here's a snippet of R code to make a plot of the points and to make a file with latitudes & longitudes..

df <- read.csv(
"http://blog.lib.umn.edu/wage0005/echinacea/sppRandCoords.csv")
plot(df$E, df$N, asp = 1, type = "n")
text(df$E, df$N, labels= df$order)
require(PBSmapping)
names(df) <- c("EID", "X", "Y") 
df <- as.EventData(df)
attr(df, "projection") <- "UTM" 
attr(df, "zone") <- 15
fred <- convUL(df, km=FALSE)
write.csv(fred, file= "sppRandLL.csv", row.names= FALSE)

Firstly, does anyone have a good acronym for this experiment? I tried PONS but don't really like it.

Also, I wrote up a list of what each person needs for next week's endeavors. If I've estimated times correctly, we need 9 people to do pollinator observations, 3-4 ppl for FNC (but the more the merrier), and 3-4 people for flagging plants beforehand (again the more the better) :ech PONS equip list.doc

What we still need :
>Ice packs (9)
>lunchbox coolers (6) I found 3 total but maybe there are more roaming around?
>stopwatch (1)--Greg offered to bring a couple back from his school for next week.Thanks Candyman!

9 possible remnants have been chosen (based on size of flowering plant populations in 2004 & 2005) and the back ups as well so here they are:
Staffanson, Landfill, North of Railroad crossing, LCW, NW Landfill, Riley, Yellow Orchid Hill, Steven's approach, Nessman. Back ups: East Riley, EELR?, Railrd crossing

Hopefully Friday pm or Monday pm we can drive around to these sites and assess the abundance of flowering Echinacea plants, and also get a good understanding of what may be co-flowering with Echinacea next week. We will need to be flexible...if there aren't enough plants flowering, then we will have to push the experiment back, and our second round later in the summer will have to be less than 2 weeks apart from the 1st round as we had originally planned.

A recent addition to our project is the potential to assess reproductive success using the style persistence method since it will be very valuable data and we are going to be collecting styles anyway for Kate.

We need to finalize protocols (I will try to post mine asap), figure out how to randomly select fl. plants in Staffanson and Landfill, the largest remnants, and I need to do some more practice runs of FNC with the newly made forms that Gretel helped me with today. We also will hopefully be able to do some practice runs of insect collecting this Friday with beemaster Amanda. I think field season is starting to get into full swing.

Thank you to everyone who has helped thus far with suggestions and advice. It has truly been appreciated.

A certain someone has thrown the digital gauntlet down, and being who I am, I cannot stand by and let that someone's remarks pass. Generations of Raths back to the Dark Ages would roll over in their graves were their descendant to back down, spineless, before a challenge. I shall outmaneuver my opponent by focusing on quality, not quantity. My posts shall be masterpieces of prose and picture, and my adversary shall soon bow down, defeated.

Today was a rather productive day, as Amy and I flagged transects in Nessman, Stephen's Approach, KJ's, Northwest of Landfill, and East Riley. They all went fairly quickly one we got the procedure down: Determine where the plants correspond to the map, lay down the first metre tape, choose and flag random spots on the length of the tape, measure outwards from that tape to the edge of the remnant, and flag that point as well. After that, we put in shiners and tags so that we could be sure of finding them again. We also flagged plants for Jennifer's and Diedre's tissue samples.

Pictures below are of:

A beautiful sunset outside Kensington. I cannot get used to the fact that the sun sets around 10 out here!

A dragonfly eating another dragonfly at Glacier Lakes National Park. This one just kinda landed on Allegra's head.

Prairie Clover! There was a lot of this at Glacier Lakes, which made the prairie that much more beautiful.

Members of Team Echinacea: Mimi, Kate, Daniel and Allegra at Glacier Lakes. Such a nice day!

Even though we were not at work, we could not stop ourselves from searching for seedlings! Didn't find any though.

There is a nest of baby birds in the Common Garden in row 41 that I found a week or two ago. There were only eggs at first, but they hatched and are now sprouting quills! Expect updates on these guys as well as regular pictures!

Note: As I was writing this, Dr. Ridley walked in carrying two Pappa John's pizzas as a break from the healthy salads we have been eating all week! A move worthy of a saint!

Hey Everyone,

I've finally made it to the flog. So excited. ^_^

For all our loyal followers, my name is Kate and I'll be starting the Masters program at Northwestern in the fall. Thus, part of my energy this summer will be devoted to thinking/researching/exploring possible masters projects. Stuart, Caroline, and Megan have already been very helpful in setting me on the right track; they all have some great ideas and thoughts on what I could focus on. I'm feeling a bit spoiled for choice, actually. Guess I have a lot of reading to do!

I will also be working with the Pollinator Sub-team of the Echinacea Team. Allegra, Amanda, Mimi and I will all be tackling various aspects of the great pollen issues surrounding Echinacea. The questions I will be attempting to shed light on include:
1) What pollen ends up on flowering Echinacea? In what quantities?
2) Is a plants floral neighborhood reflected by the pollen that ends up on the flower?
3) How does isolation impact the amount of pollen on Echinacea plants? How does the quantity/quality issue play out on isolated Echinacea vs. Bunched plants?
4) Does flowering early or late in the season have any impact on the amount of pollen a Echinacea receives?

I'll be posting a proposal type document with my methods soon, so watch for that.

Ta for now,
Kate

Potentilla arguta and other species for my project. If everyone could keep a lookout for these species, I would be grateful.

Potentilla arguta


Dichanthelium acuminatum

Panicum capillare

Potentilla pensylvanica

More spp and images to be added later.

I was recently informed that Daniel Rath has been "outblogging" me on the FLog, and I agree, he has-- but it stops now. Daniel updates the FLog several times a week, and that's cool. So from now on, every post Daniel posts, I too will post a post. Plus one.

Consider this a challenge, Daniel Rath!

To make up for lost time, and because if you're anything like my mom (and you might be my mom-- hi mom!) you love photos, here is a visual record of the past two days.

1) The past two mornings have been surprisingly cold here in K-town! Around 11:00 AM the truck bed has absorbed enough heat for a really fine cuddle.

2) On the way back from the '99 South garden, Gretal (Queen Bee) and I saw a little hummingbird trying to run with the big dogs (some swallows) atop the telephone line.

3) In the end, he was a bit of an outcast.

4) Today many of us went to the landfill to practice our independent project techniques (characterizing floral neighborhoods, catching pollinators, collecting pollen from non-Echinacea flowers, etc). I expected a dump, but I found a wonderland-- just see for yourselves!

Don't be fooled, it's not Italy- it's a DUMP. In Kensington!!

Mimi couldn't imagine what good deed could have landed her in such a place!

Then we found this Prairie Lily (Lilium Philidelphicum)

We got pulled over by this cop, and she made us characterize a floral neighborhood!

There was also some flowering Leadplant (Amorpha canescens)

I will post again soon about how my independent project plans are shaping up, so stay tuned. Don't forget to leave feedback in the comments!!

Edit: Click on the photos if you'd like a slightly larger image.