Procedure

Sample Procedure

1. Collect pollen samples from three different flowers.
2. Take fresh samples and place into dehydrator until dry.

Sem Procedure

1. When samples are dry prepare SEM stub.
2. Peel carbon tape and align with stub.
3. Place stub firmly on carbon tape and remove.
4. Take tweezers or a straight stick and divide it into thirds. 
5. Mark one third | the second third || and the third third |||.
6. Create a diagram showing where each sample will go. (Ex. |Sample 1||)
7. Place the first sample under under a dissecting microscope.
8. Locate the pollen grains under the microscope.
9. With a clean paint brush, collect the pollen grains on your brush. 
10.  Take the paint brush with the pollen grains and place it to the side.
11. Place the stub underneath the dissecting microscope. 
12. With the brush containing the pollen, carefully brush onto the stub in the correct section.
13. Repeat steps 9-14 with pollen sample 2 and 3.
14. Retrieve can of concentrated oxygen.
15. With the stub in one hand, point the concentrated oxygen 1 1/2 feet away. 
16. In a tornado motion, release oxygen directed at the stub until 2 inches away.
17. Repeat. 
18. Check under dissecting microscope to ensure pollen grains stayed in place.
19. Push stub into place of the SEM cup.
20. Turn it clockwise until it is no longer visible at eye level. 
21. Turn it an additional two notches.
22. Lift SEM door and slide cup into place until you hear a click.
23. Slide door down slowly.
24. Press maze icon to map image. 
25. Click setting> label and label selected sample With the first three letters of the sample, period and first initials of teammates. (Ex. Kanp4avmm) 
26. Locate the selected sample and zoom in at 2000x
27. If image is blurry, press contrast and auto focus until clear. 
28. Take pictures and 2000x or greater if wanted.
29. Go to archives.
30. Select 2000x picture and click ruler icon.
31. Tap one end of pollen and then the next. Record measurement. Save.
32. Repeat with 2 other samples.

Leica Procedure

1. Take first sample and place on the stage.
2. Locate pollen grain and focus. 
3. Capture picture at 35x. Repeat with other 2 samples.

Pollen Tube Growth Procedure

1. Gather pollen from one sample and brush onto concave slide.
2. Drop 1-2 drops of pollen growth media (0.1% Boric acid and 30% Sucrose) onto slide. Cover with cover slip.
3. Locate pollen under  compound microscope at 400x and look for tube growth.
4. Every 5 minutes record length of growth.  (Not all pollen will display tube growth)
5. Draw pollen or take a picture. Repeat.  

Pollen data table

Table comparison of the three pollen grains.

Recordings from SEM Microscope 12/12

12/12/13
Took photos of Schlumbergera (Cactus) and Kangaroo Paw but didn't get to take photos of Rhododendron.

cacp4ammv: Some pollen has one mark and some pollen has two marks. At 2000x the measured width is 47.8 mm. We took pictures at 5000x, 13000x and 20000x.

rhop4ammv: Took several pictures of this pollen under the label of cacp4ammv by accident, need to adjust later. At 2000x the measured width was 29.6 mm.

Background

Research on the morphology and development of pollen and tapetum has contributed to understanding the evolution of these characters and elucidating phylogenetic relationships among seed plants.
Over evolutionary time, the morphology of angiosperm pollen has evolved toward an increasing number of apertures, among other things. From a neo-Darwinian point of view, this means that (i) some polymorphism for aperture number must exist and (ii) there must be some fitness increase associated with increasing the aperture number. Pollen types with different aperture numbers often occur in the same species.
Although the majority of flowering plants achieve pollination by exploiting the food‐seeking behavior of animals, some use alternative ploys that exploit their mate‐seeking behavior. Sexual deception is currently known only from the Orchidaceae and almost always involves pollination by male hymenoptera.
Pollen grains are the carriers of the male gametes or their progenitor cell, in higher plants. They also are important tools for paleoclimatic reconstruction. They reflects the ecology of their parent plants and their habitats and provide a continuous record of their evolutionary history.

Hypothesis

We hypothesize that plants which live in dry climates produce pollen that have greater capabilities of flight, or are carried by the wind easier than pollen from plants which grow in other climates. Also, plants which live in wet climates produce pollen that is more "clingy" and have a stronger protection against the elements/environment.

If our hypothesis is supported, we expect to find pollen which live in wet climates to have more protrusions or "spikes"while ones in dry climates have a smooth outside.

If our hypothesis is rejected, we will not see these physical features on the pollen grain which help it maneuver it's environment. 

Team Agreement

Ahren: Completed Background section. Worked with SEM. Colored SEM pictures.
Virginia: Completed Procedure section. Worked with SEM. Took non-SEM pictures.
Mila: Completed Question section. Worked with SEM. Did Blog Work Checklist.
Morgan: Completed Hypothesis section. Worked with SEM. Got Schlumbergera and made table.

We all participated in looking through the microscope and conducting the experiment. We all have participated equally.

Addendum: We will all do our portions of the "Blog Work Time" Section. We shall all work equally.

Question

What is the correlation between climate and pollen type?