POLLINATION BIOLOGY LAB

BASED ON THE RESEARCH OF DR. RANDY MITCHELL

From the University of Akron

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 BACKGROUND

The interaction between bees and flowers is an excellent example of mutualism, a specific type
of symbiosis in which the interaction between two species benefits both. Bees that visit flowers gain
food (such as pollen and nectar) to feed themselves and their young, and flowers get pollinated, allowing
the plant to reproduce. Aside from these benefits, both of these parties also pay costs – the bee must
expend energy to hunt for and gather nectar and pollen, and the plant must provide those rewards. The
economics of pollination, just like the economics of humans, dictates that pollinators will make choices
among the food items available to them. The optimal foraging theory suggests that natural selection
favors individuals whose foraging behavior is as energetically efficient as possible. The basic principles
followed by the bees are just the same as those you use at the grocery store –foragers will spend the least
(in time and energy) to get the most. These choices in turn have important consequences for the mating
patterns of plants, and show that the mutualistic relationship between bees and flowers may not be as
harmonious as you might guess.
For example, from a pollinator’s point of view, moving from plant to plant is an extra cost to be
avoided (imagine that you could only buy one item at the grocery store, then had to go to a different store
for the next). Therefore, pollinators often make the shortest possible movements between flowers and
between plants. However, from the plant’s point of view, this will increase the amount of inbreeding. This
is generally bad for plants because most plants exhibit inbreeding depression (reduced growth and health
of offspring from mating between close relatives). Thus, for the plant, the ideal pollinator would visit one
flower then fly far away to visit an unrelated plant. You can see that this is not ideal for the pollinator.
This tension is an important part of pollination systems. Bees or plants that minimize their costs and
maximize their gains will have the best survival and reproduction, but bees that minimize their own costs
(e.g. moving very short distances between flowers) will automatically be maximizing the costs for plants
(increasing inbreeding), and vice versa. Keep these issues in mind when you observe plants and
pollinators today.

Lab Part I: Generating a hypothesis using field notes and observations

To gain an appreciation for the interaction between plants and pollinators, you will go to some of the
flowerbeds on campus and take field notes using one or both of the following methods:

Bee-focused observations - Record the behavior of an individual bee from when you first see it until it
leaves view. Record your descriptions (NOT interpretations) of the animal’s behavior

Plant-focused observations - Choose one or a few plants or flowers and record descriptions (NOT
interpretations) of the behavior of all animals visiting the plant.

Based on these notes and experiences, you will construct a hypothesis to explain some of your
observations and plan an experiment that will test your hypothesis.

You must turn in field notes and get your teacher’s approval for your research by the end of the day.

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INSECT IDENTIFICATION

Distinguishing bees from other insects: Butterflies are easily distinguished for most other floral
visitors, but flies and wasps can sometimes be mistaken for bees. Bees are generally fuzzier and often the
pollen they collect on their legs or the underside of their abdomen is plainly visible. Flies and wasps do
NOT have pollen baskets, and are not as fuzzy. Flies have only two wings and usually have very short
antennae. Wasps have almost no fuzz at all. Insects are small, so look carefully!
Bees range in size from <1mm to several centimeters in length. On campus today you are likely to
run into four main types of bee. Make sure you can distinguish them using the descriptions below. Color
images of these bees (and some other flower visitors) will be available in lab.

1) Bumble Bees (Bombus sp.). These bees are generally larger and fuzzier than any others you’ll
see. Each species and sex has different combinations of black and yellow fur. With a little work
you could learn to tell the species and sexes apart (there are perhaps a dozen species of
bumblebee in Ohio), but for this lab you will probably find it easier to just lump them all
together as Bombus. (You may occasionally see another large, dark bee with a yellow thorax and
a very shiny abdomen, but it is not fuzzy. This is a Carpenter bee, Xylocopa virginica; the males
have yellow faces).

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2) Honey Bees (Apis mellifera). Generally smaller than Bombus, these bees have little fur and have
a light brown and dark black coloration. You probably know them already.

3) Wool-Carder Bees (Anthidium manicatum). These are solitary bees (they don’t live in social
hives like Bombus or Apis) are about honey bee-sized, but stouter, with bright yellow and black
markings. Males are substantially larger than females, and have a prominent yellow spot on their
faces.

4) Other Bees. There are over 3,000 species of bees in North America (more than all the birds,
lizards, snakes, and mammals combined!) and this category is intended to hold them all!
For our purposes, any bee that doesn’t fit the above descriptions falls into this category. Most
will be much smaller than those described above, and are often shiny, with little hair.

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POLLINATION AND FLORAL TERMINOLOGY

Pollination occurs when pollen is moved from the stamens of one flower to the stigma
(receptive portion of the carpel) of another. This often requires that an animal visit the flower. To be a
pollinator, an animal must contact stamens and stigma during at least some flower visits.
When pollinators move pollen from one flower to another on the same plant, selfing occurs. When
pollen is moved to flowers on another plant, outcrossing occurs.
The shape and size of a flower’s petals and other structures can affect pollination. Some
pollinators prefer one type of flower, while others have different preferences. The shape and size of a
flower can dictate the amount of pollen transferred to and from the pollinator’s body. For example,
flowers in the mint family often have a “trigger” mechanism. If a bee is probing the flower for nectar at
the base of the ovary, it may contact parts of the flower that act as triggers, causing the stamens to snap
onto the bee’s head or back, dusting it with pollen.

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Flowers on a plant are often arranged in an inflorescence, all the flowers on a flowering stem, and each
plant may have several inflorescences. When a pollinator visits several flowers on the same plant (many
flowers on an inflorescence, or moves short distances between inflorescences), this can increase selfing.

Note that plants can fool you – for example, plants in the sunflower family (Asteraceae) produce
“heads” that are actually collections of dozens or hundreds of flowers!

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FIELD NOTES
Good field and lab notes are important for doing science. The goal of these notes is to help you
record your observations so that you (and others) will be able to reconstruct what you did, saw, and
experienced (with respect to your scientific goals) many years in the future. These notes are
WORKSHEETS, not finished products, and will provide the raw material from which you will begin to
work toward answers. Neatness is important in that the notes should be legible, but they should NOT be
re-written (that takes lots of time, and is prone to error). Many people have different opinions on and
methods for writing notes, but for now we’ll stick with the basics. The information below has been
tailored for field notes, although similar guidelines apply for lab work. Use pencil, since ink can smudge
if it gets wet. Field biologists often use notebooks that will fit in a pocket.

Each entry should be labeled with the following 6 items:

1. Your name, the date, and location should be at the top of the page (be specific – “Akron” would not
be sufficient, but “Ohio, Summit County, Copley HS, East side of building” is better).
2. Time of day. Include when you begin, and continue to make note of the time through the day.
3. Weather observations (e.g., temperature, humidity, cloud cover).
4. What you are observing (e.g., behavior of bees visiting Stachys lanata).
5. Method for observation (e.g., following a bee for 5 minutes, or until it is lost from sight).
6. What you observed. Sit or stand quietly and watch the bees and plants carefully! Don’t evaluate or
explain yet, just watch and record your DESCRIPTIONS of what happens.

Below your observations, label and include the following:

7. Summary of observations. After you’ve experienced the system for a while, you should record your
major impressions, and make a summary of what happened. Again, be DESCRIPTIVE at this stage –
interpretation comes later. A description is just that – a dispassionate listing of what happened,
without any interpretation of why. If you have ideas or interpretations that you don’t want to forget,
write them in the margins, and carefully label them as interpretations. This is a good place to record
which species you saw over what period of time (if you’re not sure which species, a brief description
or sketch is appropriate).
8. Your Questions and Hypotheses. Now list a few questions and hypotheses that have been raised by
your observations. For a few of these, come up with some ideas of how you could test those
hypotheses, or answer those questions using experiments or by doing further observations. You may
find it useful to have another look at the explanation of pollination biology above at this point.
Remember, good questions are cheap, but ways to answer those questions cleanly may be hard to
come by. Try to phrase your questions clearly, in ways that encourage clean answers.

If you change locations, format each entry as described above.

At all stages, feel free to record incidental observations that you feel might be important. It is acceptable
to record speculations and questions in the margin. Each time you change location you should repeat the
labeling mentioned above. Remember, write your notes so that someone who wasn’t with you would be able
to reconstruct the important features of where you were, what you saw, and what you did.

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Examples of hypotheses to test:
 Bees move farther between stems after encountering empty flowers
 Different plant species attract different insects
 Insects differ in their effectiveness in pollination (probability of contacting anthers and stigma).
 Bees visit more flowers on stems with more flowers
 Time an insect spends on a flower varies among plant species
 Time budgets (amount of time spent on different tasks) vary among insect species.

Try to come up with a workable idea that is not on this list.

Resources:

In the Library:
 Bertin, RI. 1989. Pollination biology. In: Abrahamson WG (ed.) Plant-animal interactions, pp 23-86.
New York: McGraw Hill
 Proctor, M, Yeo, P, and Lack, A. 1996. The Natural History of Pollination. Portland, OR: Timber Press.
 Real, LA. 1983. Pollination Biology. Orlando FL: Academic Press

On the Web
 iNaturalist
 leaf snap
 Bee diversity http://gears.tucson.ars.ag.gov/na/bees.html
 Pollination (focusing on plant-animal interactions) http://www.els.net/ - search for “pollination”
 Pollination (focusing on floral form) http://koning.ecsu.ctstateu.edu/plants_human/pollenadapt.html
 Introduction to optimal foraging theory (using hummingbirds as an example)
http://www.cquest.utoronto.ca/zoo/bio150y/foraging/intro.html
 More information on field notes http://bioweb.wku.edu/faculty/ameier/formats2.htm
http://members.aol.com/YESedu/collect2.html

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PLEASE SUBMIT THIS ELECTRONICALLY!
LAB PART 1:
Field notes based on observations of the CHS campus flower beds.

Based on the observations submit an electronic lab proposal that includes all the following
information.

1. State your hypothesis and the motivation for your hypothesis.

2. How will you test your hypothesis (don’t forget to mention sample size)? State the independent and
dependent variables.

3. What is the control group in your study? If no control group, explain why one is not needed.

4. What is the general protocol for running the experiment?

5. Provide the Excel (or google sheets) spreadsheet you will use to analyze your data. Make sure you
are labeling columns/rows.

4. Which possible statistical tests do you intend to do to analyze data?

5. Which type of visual data analysis do you intend to use? (bar graph, line graph etc.)

6. How will you know if your hypothesis is supported or rejected?

LAB PART 2: Testing Your Hypothesis

Once your lab proposal has been authorized (by me) start your experiment on your own time!

Follow the rubric (purple) and write up your first draft.

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Faculty Data Interpretation – 20 Points

Dr. Mitchell wanted to investigate the relationship between the number of flowers on a plant and the
number of flowers visited by various bumblebees (Bombus spp.). In his study, Dr. Mitchell
experimentally altered floral display (i.e. number of open flowers on a plant) of the square-stem monkey
flower (Mimulus ringens; Scrophulariaceae) so that plants either had two (2), four (4), eight (8), or 16
flowers per plant. To do this, he experimentally removed flowers on plants until he reached his target
goal. After plants were manipulated, he counted the number of flowers that were visited by three
species of Bombus—Bombus fervidus, Bombus griseocollis, and Bombus impatiens. As part of his data
analysis, Dr. Mitchell graphed the average number of visits (along with 95% CI) of each species when
there were two (2), four (4), eight (8), and 16 flowers per plant.

a
a

1. As the # of flowers per plant increases, how do the three species of bee respond (in general)?

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2. Explain how this behavioral response would affect the potential for self-pollination.

3. Why do bees exhibit this behavior (provide a reasonable hypothesis)?

4. One way to graphically display statistical results is to include letter notations. Letters next to
means are used to denote whether a difference exists. Means that share the same letters are NOT
significantly different from one another (so any mean with “a” is not different from another
mean with “a” but is different from a mean with any other letter). Given this information,
describe how the three species of Bombus compare in the number of flowers visited when there
were 8 flowers per plant. Your answer can include some statistical jargon but attempt to
explain the data to someone who has never had a statistics lesson.

5. How does your research compare to Dr. Mitchell’s data?

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