Analysis of Phylogenetic Trees

Our trees did not agree; our hypothesis was that the rhododendron and the mimosa were more closely related, based on their pollen structure. But based on the phylogenetic tree made from the amino acid sequences of each plant species; the mimosa and the day lily are more closely related. The rhododendron is the plant species that is least related to the other two. We trust the amino acid sequence based tree more, because their is actual scientific evidence whereas our hypothesis was mostly guesswork and just observing the pollen of the plant.

Experimental Analysis Question 

The data we have gathered has showed us that each plant leans toward more cold climates. The Day Lily originates in countries of Asia and prefers wet climates. The Rhododendron originated in Europe and North America and also prefers colder climates. The Mimosa tree, native to South and Central America, also prefers colder climates. After we shot the SEM photos and labeled the measurements we found a range of sizes between the plants. The Day Lily had a width of 59.2 the Rhododenron had a width of 42.5, and the Mimosa tree had a width of  90.3. Looking at this data the Mimosa tree is found in South and Central America which can be a hot or cold climate. If found in more hot climates the number recorded for the width could explain it's difference from the other plants. These data suggest that hotter climates increase the pollen size of the plant. In order to form a more solid conclusion, more data will be needed.

Pollen Data Table

https://docs.google.com/spreadsheet/ccc?key=0Ah_k1Z2flz9jdElqZWlRODVGcFA4VV90bVdsOGprN3c&usp=drive_web#gid=0

Tree!

Phylogenetic tree, w/ 3 specimen: Mimosa Tree, Rhododendron, Day Lily
We based our phylogenetic tree off of the region these plants were found. Primarily, we looked at the pollen characteristics of each plants. We noticed the Rhododendron and Mimosa pollen grains had folds and layers so we assumed those two plants would be more closely related. The Day Lily seemed to have grooves in the grain, unlike the Mimosa and Rhododendron pollen grains, so we placed it father away from both.

Another Phylogenetic Tree

This Phylogenetic Tree is from Jalview

Albizia_kalkora= Mimosa tree

Hemerocallis_fulva = Day Lily

Rhododendron_fortunei = Rhododendron flower

SEM PiCTURES

This is a Day Lily pollen grain with measurements. The width is 59.2 micrometers and the length is 89.5 micrometers.  

In this image, the Day Lily is zoomed in to see the outer layer of the pollen grain. 

This is a pollen grain of the Mimosa tree with measurements. The width is 90.3 micrometers. 

This image is the pollen grain of the Mimosa tree zoomed in to see the folds and texture of the outer layer of the grain. 

These are the pollen grains of the Rhododendron with a measurement on one of them. The width of the grain is 42.5 micrometers. 

Phylogenetic Tree

   Guide Tree

This is the stub that goes in the SEM. The blue lines are the fiduciary marks that separate the three pollen samples and make it easier to locate where each pollen is. The yellow circle surrounds the mimosa tree sample which is between one and two of the fiduciaries. The purple circle surrounds the rhododendron and the red circle surrounds the day lily. This picture was taken by Austin through a compound microscope.

Background Information

Generally, as temperature increases the size of each pollen grain increases as well, and changes the shape and structure of each individual grain as well.
Desiccation, or serious drying, affects the number of grains produced and the shape of the grain changes as well.

The Day Lily, originating in various countries of Asia, including Japan, Siberia, and Korea, are very at home here in Oregon, because they are used to wet climates. Though they can grow in most climates, due to the plant's adaptability.

The Rhododendron originates in Europe and North America, habitats of gardens and parks, and likes a colder climate, it has well draining, acidic soil, and the pollen is poisonous

The Mimosa pudica, is native to South and Central America. Grows in shady areas. Known for its rapid plant movement

Procedure

Using the SEM:
We were given the materials to make the stub. First we attached the carbon tape to the metal stub and centered it. Then the stub was divided into three sections called fiduciary marks using sharp tweezers. Again, using the tweezers, we transported the pollen samples off of our three plants and placed them in their sections using the fiduciary marks. In a swirly motion starting from about one and a half feet away to about two inches we sprayed canned air. The air set the pollen in place and made sure it was secure. With two fingers, we placed the stub in the cup and pushed down to make sure it was in place. Once it's flush we clicked the notch four times to the right. Then the cup was placed in the slot of the SEM until the green light turned on. Slowly and carefully, we closed the door of the SEM. The button that looks like a maze was pressed and that maps the image. Going into settings, then label and we gave a specific label separately for each pollen sample. The label said the abbreviation of the plant, the groups initials and the period number (rhodcscap2). Then back to the image, it was ready to scan and we pressed the button that looked like a plus sign and the vacuum in the SEM began. To move the image and see different sections, we touched the screen rather than moving or dragging our fingers. Using autofocus, that focuses it and you might have to use that a couple of times. The contrast helped if it was too bright or too dark to get it to the right coloring. To zoom, we turned the knob to the right and then pressed autofocus. To take a photo, press the camera button. Using a thumb drive, you transfer the pictures from the SEM to the Mac computers. To measure the images, go to archive, ruler button and then touch one side of the image and then the other of what you want to measure. You can save the measurements in a new image. Once you have completed these steps for all three pollen samples, you are ready to eject the cup with the pollen. Press the eject button, then okay and wait for it to eject. We took the cup out and twisted it so its comes up than took the stub off with out damaging it.
Stations:
During class we were in 8 stations. At each station we worked on different things like blog time, looking at the pollen from the plants we chose through a compound and 3D microscope and research on the plants. During blog time, we created the blog by using google blogger. Each person in the group has been working on different parts of the blog. When we used the compound microscope, we placed pollen from our plant samples on a slide and looked at it through the most basic microscope and took and drew pictures of what we saw. When the pollen was under that scope, we put it in growth medium. It is 0.1 germinate the pollen. In the 3D microscope, we examined it and used tweezers to break it apart and really study the pollen.