Pollination Ecology: Field Studies of Insect Visitation and Pollen Transfer Rates

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TIEE

Teaching Issues and Experiments in Ecology - Volume 2, August 2004

EXPERIMENTS

Pollination Ecology:
Field Studies of Insect
Visitation and Pollen
Transfer Rates
Judy Parrish
Biology Department, Millikin University
1184 West Main, Decatur, IL 62522
[email protected]
(217) 424-6235, fax (217) 362-6408
Tiger Swallowtail (Papilio glacus)
visiting Milkweed (Ascelpias),
© Judy Parrish

Table of Contents:

ABSTRACT AND KEYWORD DESCRIPTORS...........................................................2

SYNOPSIS OF THE LAB ACTIVITY............................................................................4

DESCRIPTION OF THE EXPERIMENT


Introduction..............................................................................................................6
Materials and Methods............................................................................................8
Questions for Further Thought and Discussion.....................................................21
References and Links............................................................................................22
Tools for Assessment of Student Learning Outcomes...........................................25
Tools for Formative Evaluation of This Experiment.........…...................................25

NOTES TO FACULTY BY AUTHOR..........................................................................26

ACKNOWLEDGMENTS, COPYRIGHT AND DISCLAIMER......................................32

CITATION:
Parrish, J. August 2004, posting date. Pollination Ecology: Field Studies of Insect Visitation and
Pollen Transfer Rates. Teaching Issues and Experiments in Ecology, Vol. 2: Experiment #3
[online]. http://tiee.ecoed.net/vol/v2/experiments/pollinate/abstract.html

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 2 Judy Parrish TIEE Volume 2, August 2004

ABSTRACT:

Students will investigate questions related to the pollination ecology of the most
common and accessible insect-pollinated flowers in bloom. Students will start with
natural history observations to answer common questions such as how long does
the flower stay open, what are its major visitors, and how often is it visited by likely
pollinators. They may then follow up the class study with their own questions, such
as whether flowers that are in large clumps are more likely to be visited than more
isolated flowers, how far the most frequent visitors fly between visits, how likely is it
that the next visit will be to the same species of flower, and whether self pollen
grows through the style more slowly than pollen from a different individual. Common
techniques in pollination studies such as determination of flowering phenology,
visitation rates, and identification of visitors and of pollen carried on visitors will be
used regardless of the question to be investigated. Spring beauties in the campus
lawn, buckeye or crabapple flowers, horticultural plantings on campus, roadside
goldenrods, or wildflowers in nearby natural areas should make it possible to
complete this lab at almost any time in the growing season.

KEYWORD DESCRIPTORS:

Principal Ecological Question Addressed: How and why are animals attracted to
flowers? How can animals and flowering plants act as selective agents upon
each other, resulting in coevolution of a mutualistic relationship?

Ecological Topic Keywords: coevolution, pollination, floral phenology, mutualism

Science Methodological Skills Developed: natural history observations in the


field, classification and use of dichotomous keys, sampling to estimate population
size (of plants, flowers, pollen in loads), defining questions, formulating
hypotheses, designing experiments, collecting and presenting data, microscope
use, graphing summarized data, and development of equations to predict
probability of visit, number of visits per flower, size of population

Pedagogical Methods Used: inquiry based learning emphasizing a specific set of


techniques (see also guided inquiry); cooperative groupwork to generate and test
hypotheses

CLASS TIME: Two hours, plus travel time if necessary, for the initial observations. An
additional hour of planning and approval of projects to address student generated
questions. At least one hour of lecture to precede the introduction of the lab.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 3

OUTSIDE OF CLASS TIME: Pre-lab questions, one hour, two to four hours for the data
collection for the class project, and two to four more hours to complete the
projects from student generated questions. Flowering phenology questions
require multiple short (ten minute) visits to the study site over the period the
flowers are open, from one day for most composite flowers up to a week for
flowers such as spring beauties.

STUDENT PRODUCTS:

• Prelab preparation of questions and definitions,


• Submission of responses to questions for further thought,
• Data set for compilation,
• Scientific research report on assigned portion of compiled class data and/or on
the student-directed question addressed,
• List of five intriguing questions generated during observations, each with a
testable hypothesis (may be part of the discussion section of the report on
compiled data).

SETTING: Outdoors, using easily accessible plants such as spring beauties in the
campus lawn, buckeyes or crabapples on campus, roadside goldenrods, or
wildflowers in nearby natural areas. Landscape plantings on campus can also be
used. Some lab/microscope work also necessary, as well as access to
computers with graphics software.

COURSE CONTEXT: This lab activity is used in four different courses at Millikin
University: (1) non-majors course, Local Flora, with 18 students, (2) freshman
biology major’s course, Attributes of Life, with five lab sections of 16 students
each, (3) upper division summer Field Ecology course for 10-12 non-majors, and
(4) upper division Plant Biology course for 12-16 junior and senior undergraduate
students. Details of how the activity is used in each of these courses appear in
the "Notes to Faculty: Uses of this Lab Activity in Different Courses at Millikin
University" below.

INSTITUTION: Private, four-year, smaller comprehensive university

TRANSFERABILITY: Useful for non-majors in local flora and field ecology immersion
classes, as well as junior and senior biology majors in upper level Plant Biology.
Also adaptable for younger students, with more emphasis on observations.
Additional comments appear in the "Notes to Faculty: Translating the Activity to
Other Institutional Scales."

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 4 Judy Parrish TIEE Volume 2, August 2004

SYNOPSIS OF THE LAB ACTIVITY

WHAT HAPPENS:

Students observe animal pollinators on flowers and work to answer


instructor-directed questions on topics such as which flowers are more attractive,
probability of visitation, and types of visitors attracted. Students may then design
an experiment or observations to test the hypothesis, analyze the data, and
prepare a formal report on their findings.

LAB OBJECTIVES:

At the conclusion of this lab...

1. students will develop an appreciation for and understanding of the importance


of mutualistic interdependence of organisms in the coevolution of structures
and behaviors,

2. students will learn to identify common flowering plants, common insect


pollinators/visitors to those plants, and common pollination syndromes
evident in those common plants and pollinators,

3. students will compile, add to, and use a data base of flowering times,
visitation rates, and a pollen reference collection usable by other classes,

4. students will learn to ask questions that generate testable hypotheses about
pollination ecology, gain experience designing experiments to test those
hypotheses, and analyze and present results in scientific format.

EQUIPMENT/ LOGISTICS REQUIRED:

• This handout and data sheets,


• Populations of flowering plants and visitors,
• Basic fuchsin gel,
• Dissecting needles for cutting and applying gel cubes, glass microscope slides,
coverslips, and candle with matches or lighter (or dark paper and sunlight),
• Permanent markers for labeling slides; slide box and small insulated ice chest to
keep prepared slides from melting,
• Microscopes with 100 power; counting grids,
• Insect nets,
• Ethyl acetate for stunning, or freezer,
• Hand lenses,
• Stopwatches or watches with second hand.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 5

SUMMARY OF WHAT IS DUE:

From this lab, students should submit the following:

1. Responses to the following pre-lab questions due at the beginning of the


first lab devoted to pollination,

• Define the following terms: coevolution, mutualism, pollination


syndrome, insect, phenology, diurnal, parasitism, angiosperm.

• Differentiate between visitation and pollination, between pollination


and fertilization, and between pollination and parasitism.

• Draw and label a “typical” flower, and describe the major functions
of each part.

• Why do animals visit flowers? Describe the characteristics of


animals that would make for a good pollinator. What rewards do the
plants provide?

• Do any visitors harm the flowers? Do any flowers harm their


visitors?

2. Students will submit data collected by their groups which will be compiled
and distributed,

3. Students will generate questions about the interactions they observed.


These questions may be developed into testable hypotheses for student
projects, or may be included as part of their discussion sections for a
research-style report using the compiled data,

4. If student generated questions are investigated, a research-style report is


required to present these data.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 6 Judy Parrish TIEE Volume 2, August 2004

DESCRIPTION OF THE EXPERIMENT

INTRODUCTION:

Why are flowers so pretty? It is of little benefit to a wild plant to be admired. Why
have plants put so much energy into the structure of flowers and production of nectar
and other rewards? Of course, the flower’s purpose is to result in sexual reproduction.
Berenbaum (1995) states that “Sexual reproduction is just as important for plants as it is
for animals when it comes to generating genetic variation, but plants have a singular
disadvantage compared to animals when it comes to sex: they can't just get up and find
themselves a mate. " Plants must rely on pollen vectors, from wind to insects to birds, to
transport their pollen to another individual. Those visitors must be attracted to the same
species repeatedly to bring about pollination. Visitors must cause pollen transfer for
flowers to be successful. Usually this means that the visitor must be attracted, collect
pollen accidentally by brushing floral parts, or purposefully collect pollen to take back to
a nest, and then visit another flower of the same species and brush up against the
stigma, effecting pollination. Flowers can attract pollinators by providing ample nectar of
the right composition, and by advertising this nectar by deep shape and recognizable
floral patterns, by providing excess pollen as food, or by providing shelter or a place to
raise (and feed) young - or by at least looking as if they do (Faegri and van der Pijl
1971). We human observers have tastes that are somewhat similar to those of the birds
and bees when it comes to floral attractiveness (although we vary from carrion beetles
and flies, as we do NOT consider the smell of rotting meat attractive). Our perception of
color differs from that of non-vertebrate pollinators. Bees don’t see colors at the red end
of what we consider the visible light spectrum, but they do see colors of ultraviolet.
Many flowers have ultraviolet markings that act as nectar guides that cue insects on
where to find floral rewards (Barth 1991, Buchmann and Nabhan 1996, Proctor et al
1996).

In some cases, there are many species of plants, or many flowers of the same
species, open at the same time, resulting in a shortage of possible pollinators and
competition among the plants for visits (Mosquin 1971, Waser 1983, Caruso 2000). This
may result in differences in flowering time to reduce competition for pollinators (Frankie
1975, Anderson and Schelfhout 1980) or in changes in floral structure (Waser 1983,
Fishman and Wyatt 1999, Medel et al 2003). In other cases, pollinators must compete
with each other, as the floral rewards are in short supply (Pleasants 1981, Pyke 1982,
Thomson 2004).

Natural selection has favored those flowering plants that are most attractive to
pollinators, and those pollinators best able to get floral rewards. Millions of years of
coevolution between flowering plants and their pollinators, with each participating
species population acting as a selective agent on the other, have resulted in
overwhelming biodiversity of both insects and flowering plants (Stebbins 1983). “The
shapes and colors of the flowers, their scent, their location on the stalks, the season
and daily schedule of their pollen and nectar offerings, as well as other qualities we
admire but seldom understand, are adjusted precisely to attract particular species of

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 7

insects; and those specialists in turn, whether beetles, butterflies, bees, or some other
group, are genetically adapted to respond to certain kinds of flowers” (Wilson 1999).
The mutualistic relationships that develop between a flowering plant , which benefits by
cross-fertilization and its most effective pollinator, which benefits from an enhanced food
source, are also influenced by other species populations. There may be herbivores and
nectar thieves that visit flowers to acquire rewards, but do not disperse pollen (Irwin and
Brody 1999, Maloof and Inouye 2000). We see the results of reciprocal selective
pressures, with adaptations of the flower to restrict nectar acquisition to a visitor with the
ability to move its pollen effectively, and adaptations of the insect to acquire nectar in
long, curved tubes. Other organisms that interact with both flower and pollinator may
complicate the system, resulting in apparent or real maladaptations (Thompson et al
2002).

The most frequent visitor to a flowering plant is not necessarily the one that is the
most effective pollinator. Natural selection will result in the flower adapting to the
pollinator that is most likely to bring about effective pollen transfer for fertilization. Floral
color, shape, placement, timing, and reward will be selected, with the most attractive
flower getting the most effective visits, producing more viable seed, and leaving more
offspring in future gene pools. The most likely pollinator can often be deduced from
floral characteristics, with flowers that attract the same type of pollinator converging in
morphology into what we call pollination syndromes (Table 1, Key 2) but it usually takes
careful observation to sort out specific pollination relationships. Most flowers attract
several different kinds of pollinators, with several different attractants which may not be
consistent with the “syndrome.”

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 8 Judy Parrish TIEE Volume 2, August 2004

MATERIALS AND METHODS:

Study Site(s): Outdoors, anywhere there are plants in flower - ranging from spring
beauties in the lawn, to flower beds on campus, to insect pollinated trees like crabapple
or buckeye.

Overview of Data Collection Methods and Analysis.

Example Assignment - Natural History Observations.

The class might be divided into three groups, one to estimate visitation rates, one
to determine the phenology (seasonal and/or daily timing of flowering) of the flowering
species (Ohio buckeye in this case), and one to collect visitors and determine how
much and what kind of pollen is carried. Alternatively, any one part could be used. Once
you have these kinds of data about the most likely flowering plants and visitors for the
time of your class, you may want to provide students with these data and go straight to
the student-generated questions. Many of the techniques are described in Kearns and
Inouye (1993).

Group 1 - Visitation Rates

Equipment Needed:

* stopwatches or watches with second hand,


* marking tape or flags,
* “eyeball ID,”
* hand lenses,
* thermometer,
* data sheets,
* tape recorder or partner to record visits so observer’s eyes can remain on
flowers.

Procedure:

1. Each person in the group should find a branch (buckeye) or a plot (spring
beauties) with receptive flowers (pollen is being shed, and/or stigmatic
surface is visible and sticky). Mark the branch or plot with marking tape
(buckeye) or flags (spring beauties).

2. Carefully examine the flowers. Use the key to pollination syndromes (sets of
traits of flowers thought to attract and/or accommodate pollen vectors, and
sets of traits of animals that allow them to exploit flowers with those traits) to
predict visitors by floral morphology.

3. Count the number of individual OPEN flowers on the branch to be observed,


or in the marked plot. Count only the flowers you will observe - and record

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 9

visits on only the flowers selected or in the plot.

4. Observe the marked branch or plot for three observation periods of exactly 10
minutes, counting the number of visits by each category of visitor (categories
include honeybees, bumblebees, small bees, flies, butterflies, beetles, and
birds).

5. Note the ambient air temperature in oC.

6. Return to the same branch or plot and make three ten-minute observations at
additional assigned time (later in the day, or earlier on another day - assigned
times will cover two hour blocks - 0800 to 1000, 1000 to 1200, 1200 to 1400,
1400 to 1600, and 1600 to 1800 hr),

7. Note the temperature in oC

8. Calculate the average number of visits per category of visitor per flower per
observation period.

9. Is the most frequent visitor the one you expected from the pollination
syndrome key? If not, why might there be a difference in what the flower
appears to attract and what actually visits most frequently?

10. What were each of the visitors doing at the flower? Does that activity
promote pollen transfer?

11. Turn in your observations to your instructor so that they may be compiled.
Pick up the compiled observations from all groups,

12. Estimate the number of visits that can be expected per flower at each
observation time.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 10 Judy Parrish TIEE Volume 2, August 2004

Report Form: Visitation Rates - Group 1, In-class Observation

Time of Day________ Temperature - Degrees Celsius________

Number of flowers observed_________ Number of minutes observed____________

Category Observation 1 Observation 2 Observation 3 Mean


Honeybees
Bumblebees
Small bees
Flies
Butterflies
Beetles
Other
Total

Report Form: Visitation Rates - Group 1, Additional Assigned Observation

Time of Day________ Temperature - Degrees Celsius________

Number of flowers observed_________ Number of minutes observed____________

Category Observation 1 Observation 2 Observation 3 Mean


Honeybees
Bumblebees
Small bees
Flies
Butterflies
Beetles
Other
Total

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 11

Key 1 - Major Insect Visitors to Flowers (based on Central Illinois)

1 Wings not visible, or hard wing covers concealing wings.... 2


2 No wings, narrow area between thorax and abdomen.... Ants
Hard wing covers conceal flight wings, form line down middle of
2' Beetles
back, chewing mouthparts....
1' Wings visible.... 3
One set of filamentous wings, eyes large and obvious (careful -
3 Flies
some Syrphid flies mimic bees)....
3' Two sets of wings.... 4
4 Both sets of wings often colorful, covered with scales.... 5
Antennae with knob-like ends, wings usually folded when at
5 Butterflies
rest....
Antennae with feathered ends, no knob, wings often open at
5' Moths
rest....
Bees and
4' Wings membranous, usually clear....
Wasps, 6
Thorax and abdomen joined by narrow “waist,” abdomen
6 Wasps
often pointed....
6' “Waist” not as marked, body usually hairy.... Bees, 7
Megachilid
7 Pollen carried on “belly”.... leafcutting
bee
7' Pollen carried mainly on leg.... 8
Halictids,
Usually small (~5-10mm), black or metallic green,
8 “sweat bees”
short tongued....
Andrenids
8' Long tongue, usually over 12 mm.... 9
Spur on hind leg, abdomen often appears Anthophorid
9
striped.... digger bees
No spur, body robust, usually over 20mm, yellow Bumblebees
9'
and black, eyes not hairy.... Bombus
No spur, golden brown color, 12-15mm, hairy Honeybees
9"
eyeballs(!).... Apis mellifera

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 12 Judy Parrish TIEE Volume 2, August 2004

Key 2 - Dichotomous key to pollination syndromes

Flowers small, inconspicuous and usually green or dull in color,


1 Wind
petals reduced or absent....
1' Flowers conspicuous, usually with white or colored petals.... 2
2 Flowers regular in shape, radially symmetrical.... 3
Flowers purple-brown or greenish in color, often with strong
3 4
odor of rotting fruit or meat, little floral depth....
4 Flowers purple-brown, sometimes with a “light window”.... Flies
4' Odor day or night, dull color.... Beetles
3' Flowers with little odor, or sweet odor.... 5
5 Flowers with deep corolla tube.... 6
Flowers red, open in day, little or no odor, no nectar
6 Hummingbirds
guide, nectar plentiful....
6' Flowers not pure red, usually sweet odor.... 7
Flowers yellow, blue, or purple, corolla tube not
Long tongued
7 narrow, but sometimes needing forced opening,
bees
often with nectar guides....
Flowers red, purple or white, corolla tube or spur
7' 8
narrow, usually lack nectar guide....
Flowers purple or pink, diurnal, upright, with
8 Butterflies
landing area....
Moths
Flowers white or pale, pendant, open or
8' (in some
producing odor at night....
areas, bats)
Flowers more dish-shaped, reward accessible, yellow, or Bees, Flies,
5'
with abundant pollen.... small moths
2' Flowers irregular in shape, bilaterally symmetrical.... 9
9 Flowers red, little or no odor.... Hummingbirds
9' Flowers with odor, usually with nectar guides.... Bees

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 13

Table 1. Pollination Syndromes: Characteristics of flowers and the vectors that


shape them.

Vector Characteristics of Flower Characteristics of Vector


Wind Inconspicuous, green or dull in Abiotic
color, petals reduced or absent,
abundant and in canopy
Beetles, flies Dull colors, dark red, strong, spicy Good sense of smell
odor, or odor of rotting flesh, flat Some lay eggs in rotting
shape flesh
May have light window (flies)
Bees Often blue or yellow, with landing Good sense of vision,
platform smell
Often have markings that act as Often have body hairs
nectar guides, sometimes in UV Can perceive depth,
spectrum “count” petals
Reduced numbers of floral parts Do not see true red – see
Often irregular in shape UV
May have deep tube or spur for
nectar
Moths Open at dusk or night, emit sweet Most active at night
odor at night Strong sense of smell
Often dull or white Have long proboscis for
Long corolla, no landing platform nectar acquisition

Butterflies Open in day, emit some odor in day Active in day


Landing platform Have long, thin proboscis
Long corolla tube, narrow for nectar acquisition
May be blue, purple, red, yellow Can see red
May have nectar guide Alight on blossoms
Hummingbirds Red, large flowers with deep nectar Vision much like human –
tube and abundant nectar see red
Little or no fragrance Long bill and tongue, large
Open in day body
No landing platform Little sense of smell
No nectar guide Intelligent – remember and
return to flowers with
abundant reward
Active in day
Approach flower and
hover

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 14 Judy Parrish TIEE Volume 2, August 2004

Group 2 – Phenology

Equipment needed:

* Colored bell wire, embroidery floss


* Flags to mark plot
* Hand lens
* Aluminum tags, or colored plastic toothpicks
* Permanent marker
* Field guide to flowering plants

Procedure:

1. Carefully examine flowers. Determine if some are shedding pollen, with


stigmatic surfaces not receptive (functionally male), or if pollen is all shed, but
stigmatic surfaces are open and sticky (functionally female), or if both
stamens and pistils are functional, or if both are finished. Note presence of a
scent or nectar. Note color patterns, and presence of nectar guides.

2. Mark at least 15 individual flowers with numbered small tags, with colored
floss, or with colored toothpicks. Note the phase or phenological state of
each. The group should decide on criteria to describe each phase (see
examples for spring beauty and partridge pea).

3. If population size estimates are required, randomly select at least 10 one meter
squared plots and count number of individual plants of the target species are in
flower.

4. Return to collect data on phenological phase of marked flowers and


population in flower at four additional assigned times or days. Observations
of class members should continue for at least 5 days, or at 4 times of day on
one day if the flowers are short-lived.

5. Turn in observations to your instructor so that they may be compiled. Make sure to
pick up compiled data.

6. Determine the sequence of each phase of floral function, the average


longevity of each phase, and how long the flower is open for visitation.

Phase Key must be designed or modified depending upon what species of


flowering plant is the target.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 15

For Spring Beauty: Key to Phases: (from closed bud to flower finished)

A- Flower enclosed in bud


B- Flower open, streaked pink, stamens very pink and erect
C- At least 2 stamens appear less pink, pistil obvious
D- Stamens folded back against petals, style clearly splits into 3 stigmas
E- Ovary swollen, petals wilting

Date and Time of Observations:

Observation Date Time


1 _______ ______
2 _______ ______
3 _______ ______
4 _______ ______
5 _______ ______

Phase at Each Observation


Flower # Phase-A Phase-B Phase-C Phase-D Phase-E
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 16 Judy Parrish TIEE Volume 2, August 2004

Group 3 – Visitors

Equipment needed to collect insects:


* Aerial nets,
* Kill jars with ethyl acetate, OR freezer,
* Insect pins and boxes,
* “Eyeball ID” for insects,
* Field guide to insects.

Equipment needed to collect pollen:


* Basic fuchsin gelatin (recipe below),
* Dissecting needle,
* Microscope slides, coverslips,
* Candle and matches, lighter, or black paper and warm sunshine,
* Marking pen,
* Microscope with 100x.

Procedure:
1. Individuals with allergic reactions to bee stings should not participate in this
portion.
2. Keep the basic fuchsin gelatin out of the sun, in a small ice chest. If phenol
was added, do not touch the gel.
3. In an area away from where the visitation observations are underway, collect
as many visitors to flowers as you can with an insect net.
4. Use ethyl acetate to kill or stun insects.
5. Use a dissecting needle to cut a 0.5 x 0.5 cm cube of glycerin fuchsin jelly.
Holding the cube of jelly on the dissecting needle, wipe pollen from the body
of the visitor. Make separate slides for jelly wiped on mouthparts, abdomen,
and legs of the insect (or other body parts as observed). Carefully remove all
visible pollen from the body section. If the specimen is a bee with a pollen
load, crush the load so that pollen grains are identifiable.
6. Place the cube on a glass slide, put on a coverslip, and gently melt over a
candle or lighter, or place on dark surface in the sun.
7. Label the slide with a Sharpee - date, time, type of visitor, body part.
8. Make a reference slide from the flowering species for comparison. Wipe a
cube of the jelly on an anther that is shedding pollen. Label it with date, time,
species, and phase of flower
9. Examine your slides under a microscope at 100X. Count the number of pollen
grains of the target species (buckeye or spring beauty) AND the number of
non-target species pollen grains. Identify non-target species if possible. Start
at one corner of the coverslip and systematically move the slide back and

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 17

forth so that the field of view covers the slide the same way that an eraser
would erase a blackboard.
10. If pollen grains are too numerous for a complete count, sample the slide by
counting 10 fields of view. Estimate how many fields of view there are (this
will differ depending on the magnification you are using). Then multiply the
average number of pollen grains of both target and non-target species
counted in your 10 fields by the number of fields to estimate the number of
pollen grains on the slide.
11. Calculate the proportion of target species pollen in the pollen load or jelly
wipe that is from each part of the body. If non-target pollen is identified,
calculate the proportion of each species pollen on the slide.

Group 3, Visitor - Target Flower Species:

Visitor # Pollen # Pollen # Pollen # Pollen # Pollen # Pollen


Species Legs - Legs - Body - Body - Mouth - Mouth -
Target Nontarget Target Nontarget Target Nontarget

1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 18 Judy Parrish TIEE Volume 2, August 2004

Additional Instructions for All 3 Groups:

Turn in the report form from your section of the investigation. Pick up the
compiled data from all groups and answer the following questions. After examining the
data, develop five questions the data raise and testable hypotheses for each of those
questions.

In addition, you may be asked to choose one of those hypotheses to to design and
carry out an experiment to test. If so, use the Format for Written Research Report below
to write your lab report.

Format for Research Report - should include the following sections:


* TITLE
o One sentence summary of the paper - should be concise and informative. Include the
appropriate taxonomic information about the organism.
* INTRODUCTION
o State the purpose of the study and enough background material to demonstrate the
significance of the study.
o This is where your hypothesis and predictions go.
o You should also refer to relevant published articles that pertain to your study (what
have other researchers learned in investigating similar topics or processes?). Start
with general background and work to your specific project.
o Citation of sources should take place within the body of the paper, right after the
information cited from that source. In science, it is important to know WHO said it and
WHEN it was said, so put the author and year in parentheses (Parrish 2004). At least
three non-textbook, published and peer-reviewed sources are required for the report.
* METHODS AND MATERIALS
o Summary of setting of study (date, time in military format - 1400, not 2:00 p.m., cloud
cover, temperature, environment of room, etc.), equipment and materials used,
information about the organism(s) studied, experimental design and procedures used,
and statistical methods.
o This section should allow other researchers to repeat your experiment.
* RESULTS
o All data and statistics. Do not include raw, unsummarized data in this section. Raw
data should be placed in an appendix. Data should be summarized and analyzed
(means, standard deviation, etc.) and presented as visually as possible. Well
constructed graphs are clearer than tables for most data.
o Graphs and drawings are figures, and are numbered consecutively.
o Tables are also numbered consecutively (separately from figures).
o Refer to the figures and tables within a narrative, describing the trends. In other words,
walk the reader through your results. The opening paragraph should state the overall

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 19

trends found in the data - but do not say SIGNIFICANT difference unless you did the
statistics to test for significance. The following paragraphs should give the specific
findings that support the overall trend. Do not say “See tables for results” or “Figure
one shows that...” or (See Figure 1). Say “There were more herbivores than primary
producers (Fig. 1).”
o The tables and figures should have headings which are clear and complete enough
that they can stand alone and the reader can extract the meaning without reading the
text.
o Any explanation, interpretation, or BECAUSE statements should not appear in the
results, but in the discussion.
* DISCUSSION
o This is the meat of the report. You should interpret your results, place them in context,
and provide supporting references.
o Compare your results to your hypothesis. Avoid statements not supported by your
data.
o You should also write about possible errors in the design and implementation of the
study. Alternative explanations for the results should also be considered here, as well
as alternate hypotheses.
o Compare your results to those of other scientists, and cite their work. Start with the
specific (your work) and then go to the general, big picture.
o The final paragraph should be your conclusions from the study. What are the main
points you want the reader to understand? Your conclusions should be forceful and
memorable.
* LITERATURE CITED
o Every article cited in the body of the paper - and none that are not cited - should
appear in this section.
o Citations are by alphabetical order of the last name of the first author listed in each
paper, then by date if you cite more than one article by the same author.
o Different journals use different formats for the literature citations, as are shown in
McMillan. To be consistent, we will use journals such as Ecology for format direction,

Juenger, T., and J. Bergelson. 1997. Pollen and resource limitation of compensation to
herbivory in scarlet gilia, Ipomopsis aggregata. Ecology 78: 1684-1695.

You should have at least three references. Textbooks and encyclopedias DO NOT
COUNT!!!! (but do cite them if you use them). Web articles may count ONLY if they
have an author - and ONLY ONE of your three required sources may be from the web.
Use current journal articles as much as possible. The quality of your introduction and
discussion depends upon good use of literature.
IN ADDITION: PROOFREAD AND USE SPELL CHECK!!!!!
o Points will be deducted for errors in grammar, spelling, punctuation, and format.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 20 Judy Parrish TIEE Volume 2, August 2004

Preparation of Basic Fuchsin Gelatin and Pollen Slides:

A jelly containing stain to make a semi-permanent microscope slide of pollen, as


suggested by Beattie, A. J. 1971. Pan-Pacific Entomologist 47: 82.

Use the following ingredients:

* distilled water, 175 ml,


* glycerin, 150 ml,
* gelatin, 50 g,
* crystalline basic fuchsin as desired - enough to make solution the “color of a
fine claret,”
* crystalline phenol, 5 g - important in humid environments, may be left out if
gelatin can be refrigerated. Don’t touch gelatin if phenol is used.

Procedure to Make the Fuchsin Gelatin:

1. add the gelatin to the distilled water in a beaker and heat until the gelatin
dissolves,
2. add the glycerin,
3. add phenol, if desired,
4. add basic fuchsin crystals a few at a time until the solution is the color
desired. Too light will not stain the pollen, but too dark may mask details of
the pollen,
5. filter the solution through glass wool or cheesecloth,
6. pour into sterile containers such as petri plates that can be covered. If
phenol is not used, refrigerate the plates and slides. They will keep about a
month without refrigeration.

Preparation of Pollen Slides

1. keep the prepared slides, and the unused jelly, out of the sun, and cool
enough not to melt,
2. with a dissecting needle, cut a small cube of the jelly out of the petri plate,
3. brush the cube of jelly against an anther containing pollen, or on the
insect body part,
4. place the cube containing the pollen sample on a glass slide,
5. place a coverslip on top of the cube of jelly,
6. gently heat the slide over a candle flame until the jelly melts. Do not
overheat, or scorch the slide. If it is warm and sunny, the jelly may be
melted by placing the slide on a dark surface in the sun instead of using
the candle flame. This will make a semi-permanent, stained specimen,
7. using a permanent marker, label the glass slide (date, species sample
was collected on).

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 21

Questions for Further Thought and Discussion:

1. Describe the phenology of the target species. Are there color changes in floral
display? If so, can you think of any advantage to the flower for the change? To
the pollinator?

2. Draw a graph to display the average visitation rate by all categories of visitors
over daily time. Are flowers more likely to be visited at certain times of day?

3. Draw another graph to display the average visitation rate by all categories of
visitor at different temperatures. How does this graph compare to the time of
day graph?

4. Calculate the probability of visit to each flower by any possible pollinator. You
will need data from the phenology group to know how long the flowers are open
for visitation, as well as average visitation rates, or rates for specific time
periods.

5. Which visitors carry the most pollen? Is that pollen likely to be transferred to
another flower of the same species? If more than one species of pollen is
present, is it in the same place on the visitor’s body?

6. Which visitors are likely to be the most effective pollinators? Why? Which
visitors are probably not pollinators?

7. What attributes of the plant population (e.g. density, patch size, identity of
neighbors) might raise or lower the predicted visitation rate?

*** Note: Answers to many of these questions and numerous other comments by the
contributing author can be found in the "NOTES TO FACULTY" section below.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 22 Judy Parrish TIEE Volume 2, August 2004

References:

Anderson, R. C, and S. Schelfhout. 1980. Phenological patterns among tallgrass


prairie plants and their implications for pollinator competition. American Midland
Naturalist 104: 253-263.

Barth, F. G. 1991. Insects and Flowers: The Biology of a Partnership. Princeton


University Press, New Jersey. 408 pp.

Berenbaum, M. 1995. Bugs in the system: Insects and their impact on human affairs.
Helix Books, Addison Wesley Publishing Company.

Bronstein, J. L., P. H. Gouyon, C. Gliddon, G. Kjellberg, and G. Michaloud. 1990.


The ecological consequences of flowering asynchrony in monoecious figs: a
simulation study. Ecology 71: 2145-2156.

Buchmann, S. L., and G. P. Nabhan. 1996. The Forgotten Pollinators. Island Press,
Washington, D.C. 292 pp.

Caruso, C. M. 2000. Competition for pollination influences selection on floral traits of


Ipomopsis aggregata. Evolution 54: 1546-1557

Faegri, K., and L. Van der Pijl. 1971. The Principles of Pollination Biology. Pergamon
Press, New York. 281 pp.

Fenster, C. B., W. S. Armbruster, P. Wilson, M. R. Dudash, J. D. Thomson. 2004.


Pollination Syndromes and Floral Specialization. Annual Review of Ecology,
Evolution, and Systematics 35 (in press, expected 12/04)

Feinsinger, P., L. Margutti, and R. D. Oviedo. 1997. School yards and nature trails:
ecology education outside the university. TREE 12: 115-120.

Fishman, L., and R. Wyatt. 1999. Pollinator-mediated competition, reproductive


character displacement, and the evolution of selfing in Arenaria uniflora
(Caryophyllaceae). Evolution 53: 1723-33.

Frankie, G. W. 1975. Tropical forest phenology and pollinator plant coevolution.


Pages 192-209 in L. E. Gilbert and P. H. Raven, eds, Coevolution of Animals
and Plants. University of Texas Press, Austin, Texas, U.S.A.

Irwin, R. E., and A. K. Brody. 1999. Nectar-robbing bumblebees reduce the fitness of
Ipomopsis aggregata (Polemoniaceae). Ecology 80:1703-1712.

Kearns, C. A., and D. W. Inouye. 1993. Techniques for Pollination Biologists.


University Press of Colorado, Niwat, CO. 571 pp.

Maloof, J. E., and D. W. Inouye. 2000. Are nectar robbers cheaters or mutualists?
Ecology 81: 2651-2661.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 23

McMillan, V. E. 2001. Writing papers in the biological sciences. Bedford/St. Martin’s


Press, Boston, MA.

Medel, R., C. Botto-Mahan, and M. Kalin-Arroyo. 2003. Pollinator-mediated selection


on the nectar guide phenotype in the Andean monkey flower, Mimulus luteus.
Ecology 84: 1721-1732.

Mustajarvi, K., P. Siikamaki, and S. Rytkonen. 2001. Consequences of plant


population size and density for plant-pollinator interactions and plant
performance. Journal of Ecology 89: 80-87.

Pleasants, J. M. 1981. Bumblebee response to variation in nectar availability.


Ecology 62: 1648-1661.

Proctor, M., P. Yeo, and A. Lack. 1996. The Pollination of Flowers. Timber Press,
Portland, OR. 479 pp.

Pyke, G. H. 1982. Local geographic distributions of bumblebees near Crested Butte,


Colorado: competition and community structure. Ecology 63: 555-573.

Stanton, M. L. 1994. Male-male competition during pollination in plant populations.


American Naturalist 144: S40-S68.

Stebbins, G. L. 1983. Why are there so many species of flowering plants?


BioScience 31: 573-577.

Thompson, J. N., S. L. Nuismer, and R. Gomulkiewicz. 2002. Coevolution and


maladaptation. Integrative and Comparative Biology 42: 381-387.

Thomson, D. 2004. Competitive interactions between the invasive European honey


bee and native bumble bees. Ecology 85: 458-470.

Waser. N. M. 1979. Effective mutualism between mutually sequentially flowering


plant species. Nature 281: 670-672.

Waser, N. M. 1983. Competition for pollination and floral character differences


among sympatric plant species: a review of evidence. Pages 277-293 in C. E.
Jones and R. J. Little (eds.) Handbook of Experimental Pollination Biology. Van
Nostrand Reinhold, New York.

Waser, N. M., L. Chittka, M. V. Price, N. M.Williams, and J. Ollerton. 1996.


Generalization in pollination systems, and why it matters. Ecology 77: 1043-
1060.

Willson, M. F. 1983. Plant Reproductive Ecology. New York: John Wiley and Sons.

Wilson, E. O. 1999. Diversity of Life. W.W. Norton and Company, New York. 424 pp.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 24 Judy Parrish TIEE Volume 2, August 2004

Links:

Pictures and descriptions of flowers adapted for various types of pollination


(wind, beetles, flies, bees, birds)
www.cas.vanderbilt.edu/bioimages/pages/pollination.htm
see also bioimages.cas.vanderbilt.edu/

Dichotomous key to most likely pollinator


www.ns.purchase.edu/biology/bio1560lab/pollination.htm

Images of 96 flowers and descriptions of their pollination


www.biology.vanderbilt.edu/BIO/review.html

Descriptions of pollination syndromes, and images


www.ftg.org/EduProfDev/Birds_Bees.html

Test of the predictability of pollination syndromes


www.umsl.edu/~biology/Bourne/resVulgare.html

Source references to Pollination biology: means of attraction, coevolution, and


diversification
www.biology.vanderbilt.edu/BIO/262sourcespollination.html

Descriptions and comparisons of Proteaceae that are pollinated by different


vectors - wind, birds, rodents, and insects
protea.worldonline.co.za/p12pol.htm

Another site on the Proteaceae


protea.worldonline.co.za/p12pol.htm

Introduction to pollination biology, pollination syndromes. Laboratory instructions


for three session investigation and presentations.
www.brynmawr.edu/biology/102_Lab03/PollinBioLabs.doc

koning.ecsu.ctstateu.edu/Plants_Human/pollenadapt.html

www.ecobooks.com/pollinat.htm

www.mobot.org/MOBOT/Research/prosoeca/discussion.html

www.mobot.org/MOBOT/Research/prosoeca/prosoeca.html

www.actahort.org/books/437/437_1.htm

www.nmp.umt.edu/geograph/edlund/g446/meeuse.html

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 25

Tools for Assessment of Student Learning Outcomes:

Guidelines for Assessment:

Each student's grade will be based on 15% from the answers to pre-lab
questions, 15% for group data collected, 10% for answers to questions for further
thought, and 10% for questions and testable hypotheses generated. The other 50% will
be from the individual research-style papers submitted (Ten points for each section -
Introduction, Methods and Materials, Results, Discussion, and Literature Cited). See
"Formal Report Scoring Sheet." A practical exam will test knowledge of flower parts and
species of flowering plants and insect visitors.

Formal Report Scoring Sheet:

Possible Points
Section Comments
Points Earned
Introduction
Background
Justification 10
Literature used
Hypothesis clear
Methods - repeatable, clear 10
Results
Tables &/or figures properly drawn & labeled
DATA PRESENTED ONCE 8
Pertinent results presented
Narrative - describes trends 1
No explanation (only in discussion) 1
Discussion
Results compared to hypothesis 3
Explanations, alternative explanations 2
Literature used correctly 2
Demonstrated understanding of coevolution 2
Strong conclusion 1
Literature Cited
At least 3 primary sources 6
Proper format 4
TOTAL 50

Tools for Formative Evaluation of this Experiment:

An extensive discussion on Evaluation appears in the Teaching section of the TIEE


website: http://tiee.ecoed.net/teach/teach.html.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 26 Judy Parrish TIEE Volume 2, August 2004

NOTES TO FACULTY

Comments by Contributing Author - Judy Parrish

Challenges to Anticipate and Solve:

Allergic reactions: Students should be asked about allergies to bee stings and
pollen. Those with potential negative reactions should be assigned to tasks that
reduce their risks.

Weather: Especially in the spring, there are days when this activity will not work
because the insects will not be active if it is raining, too cloudy, and/or too cool
(under 10oC). A back-up plan should be in place for an indoor activity that can
be alternated with this one. However, it is still possible to do the floral
assessments.

Comments On the Lab Description:

Introducing the Lab to Your Students.

I usually begin with at least one 45 minute lecture on pollination ecology and
coevolution of mutualism in the classroom portion of my courses. On the day of
lab, the approach differs in different classes. For all, we start with careful
observation of the flowering species chosen and its visitors. For the freshman
biology majors, we then divide into three groups (visitation, phenology, and
visitors) to start the natural history observations, to be compiled and followed up
on, OR we start with group observations and brainstorming questions. Students
then decide on a question to investigate in a group of about 3.

Comments On the Activities in the Lab.

Since temperature, wind, and how sunny the day is all greatly influence insect
activity, it is good to have a back-up activity that could be substituted for the
pollination observations.

Suggested Back-up Activity:

Willing, R. P. 2000. A Simulated Pollination Exercise. Pages 469-473, in


Tested studies for laboratory teaching, Volume 21. S. J. Karcher (ed.).
Proceedings of the 21st Workshop/Conference of the Association for
Biology Laboratory Education (ABLE), 509 pages.
(www1.union.edu/~willingp/ABLE/POLLEN.html)

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 27

Facilitating Development of Student Questions:

Students are sometimes stymied when asked to ask questions (even though
they were too good at it when they were five!). Feinsinger et al (1997) suggest the
following guidelines for questions:

o The question chosen should be answerable within the time allotted.

o Questions of “How? Which? How many? and Where?” are likely to be


answerable. The “Why?” questions may be more natural and more
beguiling, but are rarely answerable directly through hands-on
investigation. The other questions are likely to generate many “Whys?" for
reflection, however!

o Questions must fit the following criteria:


ƒ Intriguing,
ƒ Well-defined,
ƒ Testable,
ƒ Elements must be measurable and controllable.

o COMPARISON-TYPE questions will be easiest to address, given time and


equipment.

o Compare things that common sense and prior knowledge suggest will be
different, or where finding no differences should be interesting. Do NOT
choose questions that have obvious answers, or you won’t want to waste
your time.

o Choose a question that you find somewhat tantalizing - neither too


obvious, nor too tedious in methodology.

Hypothesis: Develop an hypothesis - a tentative explanation for what we observe

o Must be TESTABLE and FALSIFIABLE.

o Should be specific - not “temperature will affect visitation rates” but


“increased temperatures will increase visitation rates.”

o CAN DISPROVE, BUT CAN NEVER PROVE TRUE!!!! There may be


other explanations.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 28 Judy Parrish TIEE Volume 2, August 2004

Investigation: Design an experiment to test the hypothesis


o Define variables:
ƒ Dependent variable - response to treatment,
ƒ Independent variable - manipulated factor - should be only one in
order to isolate effect of variable on response,
ƒ Controlled variables - all other factors that might influence response
should be held constant,
o Outline procedure:
ƒ Treatment level,
ƒ Replication,
ƒ Record carefully steps to be performed.

o Determine controls - independent variable held at an established level, or


omitted.
o Predict outcome based on hypothesis - If...Then...

Examples of questions students have investigated:


A. SHORT TERM STUDIES
1. Do small insect visitors visit more flowers on one plant than do larger
visitors? If so, what effects would this have on cross pollination?
TECHNIQUES: “Eyeball ID” of visitors, visitation observation, data analysis.
2. Which visitors are the most “faithful” to the target species (i.e. visit only that
species)? What effects would faithfulness, or floral constancy, have on the
amount of pollen “wasted?"
TECHNIQUES: “Eyeball ID,” following visitors, collection of pollen load in
staining gel, microscopic analysis of pollen loads, sampling of pollen on the
slide, creation of pie-graph of load for each species of visitor.
3. How far between target species plants do the pollinators move for their next
visit?
TECHNIQUES: “Eyeball ID,” following visitors, estimation of distances
between target species plants visited, counting numbers of flowers visited
on each plant.
4. Where is most of the pollen carried on the insect visitors?
TECHNIQUES: Collection and preserving of insects, careful swabbing of
body parts with staining gel, sampling of pollen on the slide,
5. Do flowers in large patches attract more visitors than isolated flowers or
flowers in small patches?
TECHNIQUES: Mark and observe flowers in patches of different sizes,
“Eyeball ID,” visitation observations,
6. Do flowers with larger petals attract more visitors than flowers with smaller
petals?
TECHNIQUES: Clip petals to reduce size (nail clippers) of experimental

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 29

group, and observe flowers of different sizes, “Eyeball ID,” visitation


observations.
7. Do flowers near an artificial display of similar flowers attract more visitors
than more isolated flowers?
TECHNIQUES: Set up control and experimental plots with and without
artificial flowers of similar size, color, and shape as target species, “Eyeball
ID,” visitation observations.

B. EXTENDED STUDIES

1. Do flowers that have not been visited last longer than visited (and possibly
pollinated) flowers?
TECHNIQUES: Preparation of exclosure to prevent visitation, hand
pollination, phenology observations.
2. Do flowers in large populations produce more seeds than more isolated
flowers?
TECHNIQUES: Locate sites with high and low populations of target species,
collect seed, calculate percentage of seed set,
3. Are flowers more likely to be visited at certain times of day?
TECHNIQUES: Visitation observations throughout time flowers are open,
compilation of class data for diurnal (daily) flowering and visitation profiles.
4. What is the relationship between size of active pollinators and ambient
temperature?
TECHNIQUES: Sorting visitors into 3 size classes, observe flowers on
several different days at about the same time, measurement of air
temperature, visitation observation, compilation of class data.
5. What is the relationship between size of active pollinators and ambient
temperature?
TECHNIQUES: Sorting visitors into 3 size classes, observe flowers on
several different days at about the same time, measurement of air
temperature, visitation observation, compilation of class data.
6. What is the relationship between the relative time of flowering of an individual
in the population (early, middle, or late), probability of visitation, and seed
set?
TECHNIQUES: Mark flowers open at 3 different times within the flowering
period for that target species, visitation observations, seed collection,
calculation of seed set, compilation of class data.
7. Do pollen grains from different individuals germinate and grow at a faster rate
than self-pollen in self-compatible species? Does temperature affect rates of
germination and/or growth of pollen?
TECHNIQUES: Hand pollination, stigma dissection and staining with basic
fuchsin gel, microscopic examination of style.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 30 Judy Parrish TIEE Volume 2, August 2004

Comments On the Uses of This Lab Activity in Different Courses at Millikin


University.

This lab activity is used in four different courses at Millikin University. In a spring
semester non-majors course, Local Flora, with 18 students, we do one two-hour lab at a park
across the street when spring beauties are in flower following a two hour lecture and discussion
on pollination, including showing David Attenborough’s “Birds and Bees” video from the Private
Life of Plants series. Pairs of students spend ten minutes watching a patch of flowers, and then
brainstorm questions. We get back together as a group and share “favorite” questions, then
choose one or two to work on as a class, form a testable hypothesis, and plan an investigation
to test it. The question chosen usually deals with which flowers are more likely to be visited,
comparing color, patch size, patch position in sun or shade, or flower height. The brainstorming
and planning take about 30 minutes, and the investigation takes another 30 minutes. Each pair
of students can get in two ten-minute observation periods, which results in a sample size large
enough for statistical comparisons. We compile the data, and then students write a scientific
report on the class-generated question and experiment.

The lab activity is used differently in our spring freshman biology major’s course,
Attributes of Life, with five lab sections, each with 16 students. The lab is preceded by one or
two lectures on coevolution of plants and pollinators and a pre-lab assignment. On lab day, in
each lab, students are divided into three groups to examine the natural history of pollination of
spring beauty, crabapple, or buckeye flowers. One group assesses flowering phenology, one
group estimates visitation rates, and one group examines visitors for pollen, all using the sample
experiments included with this module. Data are compiled from all five labs and posted for all to
use. Some years students are asked to write a paper estimating probability of visitation of a
flower by the visitors most likely to be effective (based on pollen load composition). Data from all
three groups are necessary to know how long the flower is open, which visitors are most
frequent, and which are likely to carry pollen. Students are required to come up with a list of
questions generated by their observations and those of the class as part of their discussion
sections. In some years, a second lab period is devoted to designing and carrying out an
investigation of a question generated by the student, and then the write-up is of the student-
generated project rather than about the class data. When we have a data from two or more
years, such as for spring beauty and buckeye flowers, we plan to present the natural history
information in an introductory lecture/discussion on the site rather than spending the two-hour
lab collecting more of that data. We can then start the brainstorming and student-generated
hypothesis testing.

In our Field Ecology course, which is a summer immersion experience at Lake


Shelbyville, Illinois, for 10-12 non-majors, we include the lab activity much like it is used in Local
Flora. However, students have a full day to complete the investigations of questions they
generate, and can further develop hypotheses in a two-day, individual follow-up project.

In Plant Biology, a fall course for 12-16 junior and senior undergraduate students,
students work in pairs on a prairie species. One three-hour lab is devoted to observing
visitation, examining flowers, acquiring pollen samples, generating questions, developing
hypotheses, and designing an experiment to test the hypothesis chosen. Projects are discussed
and approved, and investigations are undertaken outside of class. A formal scientific report is
required.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
TIEE EXPERIMENT Pollination Ecology page 31

Comments On Questions for Further Thought:

Students often mix seed dispersal and pollen dispersal - for example, when I
ask them to bring me a flower with wind dispersed pollen, nearly all bring me a
dandelion. It is important to stress the timing and source of pollen versus seeds.

Students also often have difficulty with mathematical manipulations - even with
figuring the number of visits per flower observed during a ten-minute period. It is
often better to lead them through the development of a formula to figure rates than to
present them with one (as in the example in section 6C).

The last several questions for further thought, and the questions generated by
students, make good group discussion material.

Comments On the Assessment of Student Learning Outcomes:

Extensive notes on course assessment are in the Teaching Resources sector


of TIEE under the keyword "Assessment"

Comments On the Evaluation of the Lab Activity:

Extensive notes on how to conduct formative evaluation are in the Teaching


Resources sector of TIEE under the keyword "Formative Evaluation" and in an
Essay on Evaluation of Course Reforms

Comments On Translating the Activity to Other Institutional Scales:

Since transportation is not necessary, this outdoor activity can be adapted for
much larger groups on large campuses. Landscape beds can be used, or trees with
large, attractive flowers, like redbud, crabapple, hawthorn, and buckeye.

This activity can be useful for non-majors in local flora and field ecology
immersion classes, as well as junior and senior biology majors in upper level Plant
Biology. Also adaptable for younger students, with more emphasis on observations
and not pollen removal from insects.

Different approaches have been effective, including:

o Natural history observations as major part of lab, generating list of


questions.
o Natural history observations as one lab, with spin-off projects to address
questions raised in follow-up lab.
o Natural history information given about system chosen, with student-
generated questions addressed in experiments designed and carried out
in lab.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)
page 32 Judy Parrish TIEE Volume 2, August 2004

ACKNOWLEDGMENTS:

The major part of the idea for this lab activity was inspired by a workshop at the
ESA meetings in Albuquerque, New Mexico by Alan Berkowitz and Kathleen Hogan,
1997 on Schoolyard Ecology for Elementary School Teachers (SYEFEST), and by
Peter Feinsinger’s work with Berkowitz, Margutti, Grajal, and Oviedo. Much of the
design of the activity was done as a module that was a part of a National Science
Foundation grant (DUE #9653676) to Millikin University, “Creating Linkages Through
Institution Wide Reform of the Science Curriculum.” Dean Mauri Ditzler and Clarence
Josephson, of the Millikin Chemistry Department, and Marianne Robertson of Biology,
encouraged the development of the module. This submission has benefited from
comments by TIEE Editors and an anonymous reviewer.

COPYRIGHT STATEMENT

The Ecological Society of America (ESA) holds the copyright for TIEE Volume 2, and the
authors retain the copyright for the content of individual contributions (although some text,
figures, and data sets may bear further copyright notice). No part of this publication may be
reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic,
mechanical, photocopying, recording, or otherwise, without the prior written permission of the
copyright owner. Use solely at one's own institution with no intent for profit is excluded from the
preceding copyright restriction, unless otherwise noted. Proper credit to this publication must be
included in your lecture or laboratory course materials (print, electronic, or other means of
reproduction) for each use.

To reiterate, you are welcome to download some or all of the material posted at this site
for your use in your course(s), which does not include commercial uses for profit. Also, please
be aware of the legal restrictions on copyright use for published materials posted at this site. We
have obtained permission to use all copyrighted materials, data, figures, tables, images, etc.
posted at this site solely for the uses described in the TIEE site.

Lastly, we request that you return your students' and your comments on this activity to
Susan Musante ([email protected]), Managing Editor for TIEE, for posting at this site.

GENERIC DISCLAIMER

Adult supervision is recommended when performing this lab activity. We also


recommend that common sense and proper safety precautions be followed by all participants.
No responsibility is implied or taken by the contributing author, the editors of this Volume, nor
anyone associated with maintaining the TIEE web site, nor by their academic employers, nor by
the Ecological Society of America for anyone who sustains injuries as a result of using the
materials or ideas, or performing the procedures put forth at the TIEE web site, or in any printed
materials that derive therefrom.

© 2004 – Judy Parrish and the Ecological Society of America.


Teaching Issues and Experiments in Ecology, TIEE Volume 2 (tiee.ecoed.net)

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