Recently in Pollen Interference 2009 Category

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Hey team,

Things have settled down a bit and I've started work again on the great pollen challenge! I have ten locations for each of ~150 slides, and for each location I have been recording the count of pollen grains, as well as the number of species as best I can tell (I have also taken notes with descriptions of pollen in each location). My goals at this stage are to get better at recognizing pollen grains of the same species in multiple photos and to get a feel for the diversity and amount of pollen on the pollinators we caught. I'd also like to see if there's any pollen load size/diversity consistency within a pollinator species.

I have started looking at the male Melissodes sp. and so far it looks like about half of them carry no pollen at all, but some of them have multiple grains at each location.

My question for you is... What makes an insect a 'pollinator' in the context of this study? We are focusing on pollinators, and are not including insects that we caught but know are not effective pollinators (ex. syrphid flies), so there needs to be some way to distinguish between other effective and non-effective pollinators. I have thought about making a cutoff like, say, in order for an insect to be a 'pollinator' it must have one grain of pollen per location. That way insects that happen to be carrying one grain of pollen (total) but that aren't really pollinators wouldn't be counted as pollinators in this study. However, any cutoff seems very arbitrary. It almost seems better to include anything that we know carried pollen, even one grain.

But what about those male Melissodes sp.? If some individuals carry no pollen and others carry quite a bit, do they all count as pollinators, or just the ones that carried pollen?

If you have any ideas, please put them in the comments!


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DATA

Here is a copy of my excel file with all my data. The sheet labeled corrected data for analysis is the file with all the data for each treatment. The sheet labeled baggins has a list of all the heads I used that need to be harvested, this was also posted in a separate post called "Baggins." The ones labeled "donor" do not need to be harvested but should not be expected to have high seed set as they were bagged for most of their flowering time.

Pollination interference data (data entry 2).xls

HARVESTING

I need all of the heads used in my project (the ones that are painted) to be harvested in egg cartons (located in the shelf above the sink in the Hjelm house). Please label the compartment of the egg carton with the row, pos and tt color of the head. The egg cartons should be packed so that they wont be disturbed during travel and please be careful with the heads so no seeds fall out! Gretel has a list of all the heads to be harvested and its posted on the flog in July ("Baggins"). Please mail the packaged heads to: 119 School Street, Keene, NH 03431

LANDFILL SPECIES

I have been collecting plants at Landfill this season and have started to compile a list of the plants I have either seen or collected there. If Megan and anyone else would like to add plants to that list that would be great! Or check my identification. This list is very rough at the moment but I will continue to update it as I get more plants identified.

Species list landfill 2009.xls

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Howdy gang!
Here's the list of tag ID's and corresponding letters at the sites used this summer. We flagged different plants on 7/13 and 7/6 and used the same plants for observations on 7/21 and 7/23. For some reason I can't find the list of flagged plants for 7/13, so it would be great if someone could check on the Hjelm house computer for that info. It may or may not be in the folder for this experiment. I'm sure I compiled that info from the visor memos, but I don't have the file on my computer.
ech flagged plants and tags.doc
We recorded the tag ID's during FNC so we could go back and check to make sure we had recorded the right number, but we never made the check.
Here's the file that lists whether the vial had a bee, fly, bfly, or beetle in it:
ECH poll obs ALL.xls
Here's the FNC Data in an excel file:
ECH FNC.xls

I hope everything's going well in MN! It sounds like lots of progress has been made since I left. I thought my poster presentation went pretty well back in Chi-town. The final version of it is in a previous flog post. Thanks again to everyone...I certainly could not have done this without all of your help. I hope the field season ends well. Keep in touch. Oh and here's an interesting paper I came across recently: brown bj loosestrife comp.pdf
And I really like this picture Daniel took:
P7300552.JPG
And here's Echinacea taking center stage at CBG:
P8060614.JPG
P8060592.JPG

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Here is a file with the pollen storage data (excluding Stuart's data on the 48 hour style persistence).

pollen storage data sheet.xls

We pollinated 3 plants in the common garden that were still flowering. Each plant had 3 treatments of stored Echinacea pollen; ambient temperature, frozen, and refrigerated. The frozen and refrigerated pollen caused shriveling, the ambient temperature treatment did not.

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Hey All,
Long time no post, I know. Things certainly have been busy around here. As you all probably know, I finished making slides a while back - 372 slides total. WooHoo!!

Now, onto the next step, taking pictures of these slides. I took my very first pictures today, just a couple to get the hang of things, they are attached to this post. From my fiddling around today, I can see that this is going to be a lot more work than I thought. First off, the pollen is hard to find, it's not all at the same level of view, some of them are on top of the stigma and then they're really hard to see. Also, it's challenging to focus in enough to where I can ID pollen grains. Stuart suggested working on a random sample of my slides for the rest of the summer, and completing them this fall at the CBG. The only issue there is the change of machinery, but hopefully we can figure something out.

As for the rest, I think I'll be taking 1-2 shots of the entire style, and labeling them thus: stylevialID_site_date/time_A# - so A1, A2, etc. Then I'll zoom in and proceed to take pictures at sites B through F on the style, and for each change in focus will be another number. The question is whether I should attempt to get a good sense of the exact number of pollen grains on the stigmas or try to ID the pollen types. I'd like to be able to do both, but I think for this summer at least, I'll try to get a handle on the former rather than deal with the later.

On that note, Caroline suggested using a clicker to count the number of pollen in a picture, and that seems like an excellent suggestion. Does anyone know if we've got one?

Anyway, enjoy the pictures that I took so far:
737BA_SPP_11am_20JUL.jpg

416ZU_LC_23JUL_A.jpg

416ZU_LC_23JUL_B.jpg

-Kate Monster

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Here is my dataset that I am working on analyzing in R as a .csv file.

halverson.data.091.csv

Stuart, here is my R script so far:

halverson.data.analysis1.R

I made new columns in the .csv spreadsheet for the factors and levels we discussed. I will work on a list of hypotheses to test. I think I changed the definition of "y" when I did my 24 hour analysis. Can I give "y" a different name for each analysis? Or does the code need to read a defined "y" each time?

Thanks for the help and check out the graph of 24 hours and the summary m2.

Allegra

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Here are the data on the three pollen types and the protocol for measuring.
I used the same plant/pollen from each plant and measured at least 30 different pollen grains from each. I didn't use any pollen if its pole faced forward - only if it was sideways.

Bad news - the Ech. ang. and Heli. heli. are very close. Good news - maybe the pollinators and plants can't tell them apart either.

Book1.xlsx

PS - I am in SE Minn and the Monarda Sunflower and Miss. Goldenrod are in full bloom all over.

Protocol for slide image recording and measuring.docx

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Until I get the website done, this list of measurements is all I can offer you. The measurements are in pixels. The measurements were made from multiple pollen on a single slide.Slide list.xls

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Perplexed by Stuart's question - a trip to the hilltop here in Watertown, and Mimi's poster, I checked again on the amorpha pollen - it is NOT bean shaped. But I do have reliable pictures - (Amanda don't bother getting its pollen tomorrow)

What keeps amorpha and medicago sativa from occupying the same locations? Legume wars underground? Does Andrea have insight?
Amorpha canescens g.jpg

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Here's a snippet of R code showing how to extract info from the shrivel character data (a file is below...
df <- data.frame(shrivel.txt =c("x", "xoxx", "xxxx", "oooo", "xoooo"))
df      # start off with this data frame
str(df)

df$shrivel.count <- nchar(as.character(df$shrivel.txt)) #add column

vx <- gsub("o", "", df$shrivel.txt)  # replace o with ""
vx
df$shrivel.xs <- nchar(vx)           # make a new column in df

vo <- gsub("x", "", df$shrivel.txt)  # replace x with ""
vo
df$shrivel.os <- nchar(vo)           # make a new column in df

str(df)
df      # final data frame
codeForAllegra.r
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2 graphs that are basically the same as the one with alfalfa on my poster, bu tusing sweet clover and amorpha instead. Although amorpha is the most common native species in floral neighborhoods per unique plant, there are only 43 plants that had either just amorpha, just echinacea, both , or neither. For sweet clover, the sample size is 82, but the graph isn't very impressive either....

Do you think either one is usable? I originally wanted to use the most common native and the most common exotic (alfalfa and amorpha).
ecan amca meof comparison graphs.xls

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Hey yall,
Back in Chi town finally after about 10 hrs of traveling. I said "Get er done" to someone today out of habit and got a weird look. Thanks guys. I'm also going through baked goods and other delicious foods withdrawal.

Final version of poster: Jenkins REU09.pdf

Hope everyone's doing well in K-town! I miss yall already...and my bike :(

ps. I knew Roxy would come through for us. Warren should've known better after the snake incident.

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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

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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.

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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.

P7300576.JPG

P7300578.JPG

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

P7300584.JPG

Thanks to Mimi for letting me use her camera!

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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...

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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.
P7151455.JPG

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 theseYL1304N119-5b.jpg:

LS1212J124-4.jpg

LS1212J124-9b.jpg

3) I've pinned each specimen with a unique ID code that corresponds to its vial ID number.
P7281460.JPG

P7281461.JPG

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

Male Melissodes sp.
P7281462.JPG

And Ceratina sp.
P7281463.JPG

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

-Amanda

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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... ^_^

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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!


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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...


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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

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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

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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.

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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.
P7110320.JPG
P7110309.JPG
P7110318.JPG
P7110312.JPG

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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

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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.

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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:
P7080213.JPG
Here's some pictures of our fun 4th of July and the amazing sustainable sandcastle.
P7040196.JPG
Waniel, Per, and Hattie
P7040194.JPG
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.
P7100260.JPG
A flock of pelicans flew overhead at NWLF.

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

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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

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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.

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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

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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

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RESULTS FOR NAME THAT POLLEN

Slide 1: Heliopsis

Slide 2: Coreopsis

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

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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.

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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

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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!

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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

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0001.jpg

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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!

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

echinacea03July2009.jpg

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

Love,
Daniel and Amanda

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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

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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.

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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

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My project is almost ready to start this week!

I will be hand pollinating heads on 20 flowering plants in the '96 section of the CG with different species of foreign pollen and Echinacea pollen to simulate pollen loads arriving from generalist pollinators. The Echinacea pollen will be applied at the same time as the foreign pollen, 4 hours after, or not at all depending on the treatment. I should start painting bracts for the first flowers in the next couple of days. After discussing the pros and cons of different pollinator exclusion methods, I have decided I will use bags for the 20 plants in my study, and cages for the plants I will be taking pollen from frequently. All my plants now have blue flags in the CG. Ruth brought new paint for us to use, including a lovely new dark purple color. Thanks Ruth!

I have also decided to use 3 different foreign pollen donors and no mixes for my treatments. After some practicing collecting pollen from different species at the Landfill, here are the best species I have come up with that have easy to collect pollen:
Lead plant-Amorpha canescens-Fabaceae
False sunflower-Heliopsis helianthoides-Asteraceae
These are both common in the remnants and co-flower with Echinacea, hopefully sharing pollinators.

Other possibilities:
Coreopsis palmata-Asteraceae
Thistle-whichever is in flower?-Asteraceae
Prairie anemone-Anomone sp.-Ranunculaceae

Also, the bike gang is now 7 deep, so K-town betta watchout!

Allegra

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Here's a file that lists sites you can choose for your study.

Also, here's a list of equipment that we used for during the first field season where we did systematic observations and collections of pollinators.

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Here's a file with plants that Allegra can use for her pollen interference/precedence experiment. These are all plants identified yesterday as going to flower in 2009. Plants are sorted according to priority order--random, except that plants from site of origin "Unknown" are given priority 1.

I recommend going down the list and excluding all plants that don't have at least two promising heads. Flag all included plants with labels 1, 2, 3, .... Stop when you have enough for your experiment. Ask Gretel which color flag to use. Blue is an option because your plants are in CG96 and Andrea's are in CG97.

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Here's what I've come up with for the revisions to my original proposal as of Friday's group discussion; it is not in full form yet but I wanted to flog what I have so far so that people could read it and correct any mistakes I've made, make suggestions, etc.

Thanks
jenkins echinacea proposal revised.doc

Oh and friday was a big day in the common garden because the first Echinacea plant (in the 99garden I believe) released its pollen. Tomorrow we will investigate to see if any more have followed suit. We also searched for spittle on Echinacea plants for an hour on friday to help Daniel know whether he has a sufficient sample size and found 22 with spittle on them in roughly half the garden.
And I like pretty pictures so here's one for fun
P6200016.JPG
-Mimi

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Here's my proposal for my project. Read it. Savor it. Constructively criticize it.
jenkins echinacea proposal.doc

It's still very helter-skelter at this point and in need of much fine-tuning, so any suggestions are greatly appreciated. Thanks!

Also, I'll re-attach the docx files from my last post in doc format.
Echinacea Pollinators nesting2.doc
Protocol for Taking Pictures of Insect Specimens.doc

On a side note, yesterday was a really exciting day because I found my first seedling, we got two bikes at a garage sale for $25 each, and there were the Runestone Days fireworks in the evening. The party lasted long into the night in K-town, and I think I remember falling asleep to the sweet sounds of AC/DC You shook me all night long coming from the street dance. These folks know how to party. I'm looking forward to the kiddie parade tomorrow! Although Amanda and I were saddened to hear it wouldn't be a kitty parade.

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Hello again, field log readers--

I know it's been a trying twenty hours of waiting, but I now have more details regarding my independent project!

GallinatProposal.doc

Just by clicking on my proposal (above) you will get an idea of the specific questions I'd like to answer, how I plan to go about answering those questions and how my study fits into this summer's bigger picture work on competition for pollination. Take it from me, it's a riveting read!

I would greatly appreciate any questions or comments about this proposal-- whether you are part of the Project or just an Echinacea enthusiast in K-town for Runestone Days, feel free to write in the comments. Thanks!

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My name is Allegra Halverson and I am from New Hampshire. I am an undergraduate student in Botanical Science at McGill University in Montreal, and a recent addition to Team Echinacea. Lots of things happened this week, so here are a few highlights:

We moved into the old town hall and I've been loving the bike ride to the farm in the mornings so everyone with access to a bike should bring it!

I saw a garter snake, two frogs, two deer, ground squirrels, a wild turkey and lots of birds.

Gretel and I selfed Megan J's prairie turnip plants at the landfill site on Wednesday. We also helped Andrea put out flags and fungal traps in the CG for her mycorrhizae project.

I started my plant collection at the landfill and common garden with 15 plants so far. I have to make a plant collection for a class next winter and will also make one for the Echinacea project at the same time to help future newbies with plant identification.

During this first week we received a lot of background information on the project and began the planning stages of our own projects related to the larger questions about Echinacea in the fragmented prairie habitat. Several projects surrounding the question of competition for pollinators were chosen along with pollen identification projects and one project about the aphids. My project will focus on how inter-specific pollen landing on Echinacea flowers effects style persistence. pollen competition proposal.doc

We developed a new key for the labeling seedling search maps:
-each plant in the circle has a dot with line drawn to the center and the distance (cm) to the focal plant written on the line

s with a circle around it: a seedling
B with a circle around it: a basal plant, not flowering
* with a circle around it: a flowering plant, should have a metal tag like this 7819.2 (.2 is the number of flowering heads)
N with a circle around it: a nail with a metal tag on it

any plot with a plant found in it, other than the focal plant, had a map made for it.
any plot with a seedling found in it was photographed and a pencil marker with a letter (for basal or seedlings) or number (for numbered plants) was placed 2 cm west of all plants
a toothpick was placed 5 cm from the seedling towards the focal plant

am i missing anything?

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