What in the World is Wolbachia?

For our very last AP Biology lab EVER, my class looked into a very interesting type of bacteria: Wolbachia. Wolbachia is a genus of intracellular bacteria that lives inside the bodies of arthropods and nematodes. Wolbachia bacteria must live and reproduce inside the of insect host cells, as they are endosymbiotic. What exactly does Wolbachia do? Well that is both simple and complicated – Wolbachia bacteria alters the sex of their hosts, usually from male to female.

When I first heard what Wolbachia did, I genuinely could not believe it. How could a bacteria alter gender?? After doing some research, I discovered a few different ways:

1. In some species, infection of Wolbachia bacteria can cause females to reproduce parthenogenetically, meaning that eggs can develop without male fertilization. This would mean that the resulting offsprings would be genetically identical to the mother; they would all be female as well. Wolbachia causes the cells to stop mitosis at anaphase, when the two sets of chromosomes are suppose to separate, which leads to diploid nucleuses. This does not allow the duplicated chromosomes to separate, and thus results in a diploid female.

2. In some species, Wolbachia infections causes fertilized eggs to always develop as female by suppressing the production of masculinizing hormones. This makes it so genetically male embryos develop as females.

3. In some species, Wolbachia causes male embryos to abort in early development. This does not directly change a particular insect’s gender, but it does contribute to the overall feminization effect of Wolbachia.

As you can probably infer, Wolbachia bacteria reside in the reproductive systems of arthropods. Something even more interesting about Wolbachia is that it can be transmitting two ways: horizontally through contact with another organism or the environment , or more commonly, through the female’s eggs. Males cannot pass the bacteria to their offspring due to the absence of eggs in males. As mentioned previously, Wolbachia is a parasite in arthropods. Parasitism is when one organism benefits at the expense of the other – in this instance, the Wolbachia lives and reproduces inside of the reproductive organs of insects and some other arthropods, but causes an effect to the body. Interestingly enough, Wolbachia isn’t always considered a parasite: Inside filarial nematode hosts, Wolbachia is mutualistic – both organisms benefit.

Some of our Lab Equipment
Some of our Lab Equipment 

For this lab, we all went home and captured insects to use for samples and data. The purpose of this lab was to determine if any of these insects have Wolbachia DNA. This lab was like a master lab, involving many types of biotechnology that we have experimented with over the year. We began by crushing the insects’ thoraxes (where their reproductive organs lie) in a lysis solution. We then added DNA primer and a “master solution”, which contained free nucleotides. We ran these samples in a thermocycler to conduct PCR. The process of PCR (polymerase chain reaction) is used to amplify two regions of the DNA so that we could later successfully isolate the Wolbachia data. After the PCR, we began gel electrophoresis to see the results of these amplified products. The presence or absence of the genes on the gel served as date for us to determine whether or not the insects were infected with the Wolbachia parasite. The results of the gel electrophoresis are shown below.

Wolbachia.005

I (Marissa) worked with Davis and Elina, and out of all of the groups, ours seemed to be the most successful in getting accurate data. From our results we can conclude that my insect (found in San Francisco by the other biology teacher) did not contain Wolbachia DNA while Davis’ did. As you can see, only one other person seemed to find Wolbachia DNA, and many other groups were not able to successfully extract insect DNA at all.

Me holding my Data Sample!
Me holding my Data Sample!

I very much enjoyed this lab. I will admit, there were a lot of steps, and some of it was extremely confusing. However, I could definitely tell that this was a very good lab to review all of the technology and lab experiments we had done all year. We used micropipettes, gel electrophoresis, and various machines to conduct PCR. In a perfect world, there would have been more data from the rest of our class so that we could tell the frequency of Wolbachia in our area. I think Wolbachia is an incredibly interesting genus of bacteria, and I am glad that we got to learn about it at all. I was very proud of myself for participating with the micropipettes on this last lab – usually I am too shaky to do it, but this time I was able to relax and compose myself in order to do it. What a great lab to end the year with!

Kingdom Protista

This week in AP Biology, we performed a lab to learn more about organisms that fall under the Protist Kingdom! We examined organisms under a microscope, created sketches, conducted research on each individual protist, and created a group project on the Kingdom Protista. Additionally, we did some brief research on the three main types of seaweed: red, brown, and green. I worked with Allie Coon and Jennifer Schulz, and we created a prezi for our visual presentation. Click here to see and interact with this prezi!

Screen Shot 2015-03-08 at 10.33.48 PM

Bacteriophage Therapy

My partner Abi Grassler and I made an visual infographic page on http://www.easel.ly about bacteriophage therapy. Bacteriophage therapy is a process in which phages can be inserted into cells in order to target a specific strain of bacteria and either change the bacteria or expel it from the organism. We focused on the benefits of bacteriophage therapy over antibiotics, which we depicted in a chart. We also provided two diagrams of a bacteriophage and bacteriophage viral replication. Check it out here!

Screen Shot 2015-01-29 at 12.15.04 PM

Ebola and IDEO Challenge

We have been researching ways to help fight Ebola both inside and outside the classroom. For a project in class, I created an iMovie to educate others about the Ebola Virus:

We also participated in the IDEO challenge, which is an online challenge to contribute ideas to fight Ebola or at least suppress it. Here is a link to farther read about my idea to fight Ebola by fighting dehydration!

https://openideo.com/challenge/fighting-ebola/ideas/well-well-well-let-s-cure-ebola

What are your ideas to cure Ebola?

Making Meiosis Move

I collaborated with Abi Grassler to create a moving project that demonstrates the steps of meiosis in gamete cells. We used craft supplies – mainly Pipe Cleaners – to symbolize various organelles and depict the changes made and steps taken during meiosis. We then took pictures on an IPad, and used the Stop Motion application to string these pictures together in a motion film. You can view this video below:

Meiosis is a form of sexual reproduction and cell division. Unlike mitosis, which creates clones of the parent cell, meiosis creates genetic diversity – especially through crossing over, which occurs during Prophase I. Meiosis is similar to mitosis in that the cell goes through similar phases in both processes. However, meiosis goes through two divisions, producing four daughter cells, as opposed to mitosis which divides only once and produces two.

While Abi and I enjoyed using Stop Motion and taking pictures to create an animation of meiosis, using the craft supplies was challenging. Both Abi and I were originally unclear about the general process of meiosis, and we found that this confusion (on top of the struggle of using Pipe Cleaners and the time crunch) resulted in a lot of difficulties. However we did our best, and we are both really happy with the outcome of our project!

Work Cited:

Reece, Jane B., Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, and Robert B. Jackson. Campbell Biology. AP Edition ed. Vol. 10. Boston, MA: Pearson Learning Solutions, 2014. Print.

The Miracle of Mitosis – Cell Division

Title: Examining Cells from the Tip of an Onion Root to Determine the Time the Average Cell Spends in Each Phase of Mitosis

Purpose: The purpose of this lab is to determine how long cells spend in each phase of mitosis

Introduction: Mitosis is the process in which a cell divides, splitting the nucleus, and creating two identical daughter cells. Mitosis can be broken down into five major phases: Prophase, Prometaphase, Metaphase, Anaphase, and Telophase. The cell spends most of cell life in Interphase, however, which is a time for cell growth and DNA replication. Interphase takes part during the G2 cycle. During Interphase, a nuclear envelope encloses the nucleus, which contains one or more nucleoli/nucleolus. The cetrosomes have duplicated, and chromosomes cannot be seen yet. The chromosomes begin to become apparent during Prophase, the first phase of cell division, where the chromatin fibers condense and tightly coil. The nucleoli disappear during this phase and the mitotic spindle begins to form. Though prometaphase is the next phase, where the nuclear envelope begins to disintegrate and the kinetochores attach to the centromere, for the sake of this lab we did not identify this phase because it is so short and hard to distinguish. Next, during metaphase, the nuclear envelope completely dissolves and the centrosomes are now at opposite polls of the cells. The chromosomes line up in the middle at the metaphase plate, and the sister chromatids are attached to the kinetochore microtubules coming from opposite poles. Anaphase comes next, which is when the spindle fibers begin to shorten and the two sister chromatids of each pair start to part, moving toward opposite ends of the cells. The cell begins to elongate, and by the end of anaphase the two ends of the cell have equivalent and complete collections of chromosomes. The last phase is telophase,  which is when two daughter nuclei form in the cell and the envelopes begin to reform. The nucleolus split, and the cleavage furrow begins to form. After telophase is cytokinesis, which is not technically a phase because cytokinesis is the splitting of the cell, and mitosis refers to the division of the nucleus. Each stage of mitosis takes different amount of time, and while the observation of a complete cell cycle (24 hours) under a microscope is impossible, scientists can get a idea of the time spent in each stage by studying an onion root cell, and counting how many cells within the onion root are in each stage, thus creating an average percentage of cells per phase.

Method:

1. Take a sample of an onion root tip and put it under an electric microscope to observe.

2. Count and record how many cells there are total in the section of the cell you are looking at.

3. Identify which phase each cell is in, and record the data.

4. Once you have a count of how many cells are in each phase, divide the cells in each phase over the total amount of cells to find the average percent of cells in each phase. Then, by multiplying that percentage by the number of minutes in an hour (1440 minutes), find the amount of time the cells spend in each phase.

5. Record all date. Compare the percent of cells in each stage to find approximately how much time each phase would take by looking at the percentages relative to each other.

Data:

Screen Shot 2014-11-02 at 9.58.18 PM

Here is a picture of the Onion root tip that we took through the microscope
Here is a picture of the Onion root tip that I took through the microscope

Possible Errors:

Because we are fortunate enough to live in a age of advanced technology, I was able to use my phone to take a picture through the microscope. This thus magnified my view of the onion root tip, lowering the cell count. While this made the cells easier to observe/manage, but minimizing the amount of cells I now realize I also minimized my data accuracy. Therefore, the percent of cells in each stage, as well as the time a cell spends in each phase, is more of an average. The more data one has the higher chances of accuracy, and because I only had two points of data that also takes away from the accuracy of the results.

Works Cited:

“Online Onion Root Tips.” Online Onion Root Tips. The Biology Project, n.d. Web. 02 Nov. 2014. <http://www.biology.arizona.edu/cell_bio/activities/cell_cycle/cell_cycle.html&gt;.

All About Ebola

This is a video about the background of Ebola: where the disease originated, what it does, symptoms,etc. Here are the basics about a disease that has wrecked havoc and elicited fear from hearts everywhere:

In order to understand a disease, one must understand the logistics of the virus and what is happening on a molecular level. Here is a link to Amanda’s video, which demonstrates the process behind cell signaling:

To learn more about Ebola on a quantitative level, watch Davis’ video which includes data, graphs, and statistics:

Lastly, to find out what scientists are doing now about the current Ebola outbreak, view Abi’s video to learn about current research:

Reflection:

I believe that many people simply look at the fatality rate of Ebola and blow it out of proportion as an “unsolvable monster disease”, forgetting that it follows the basic behaviors of the typical virus. If one were to simply break down the virus and exactly what it does (both the steps and the effects), it makes Ebola a little less scary and the possibility for a cure seem more plausible. I really did not know what Ebola did to the white blood cells or why the bleeding/hemorrhaging occurred. Doing all of this research also led to my connection of fatal diseases to deeper, root problems, such as poverty, sanitation, and water accessibility. By looking at the differences between the death percentage in first world and third world countries affected by ebola, I understand that while a disease can affect anyone, the conditions of the area it affects can determine how detrimental it will be. This motivates me to want to understand what these core problems are and how I (and others) can help.

Works Cited:

“Ebola Virus Disease.” WHO. N.p., n.d. Web. 05 Nov. 2014. <http://www.who.int/mediacentre/factsheets/fs103/en/&gt;.

Chromatography Lab

Chromatography is the process of physically separating substances in order to be identified and analyzed. The major factor that affects chromatography is the type and saturation of the solvent being used. The dimensions of the chromatography paper, temperature of the solution, and particle size of the solution all act as factors on the rate of chromatography as well. In our particular lab, we focused on the chromatography of pigments in leaves. The purpose of the chromatography paper was to distinctly show the separation of pigments, and the purpose of the solvent is to move the pigments up the chromatography paper through capillary action in order to separate the pigments. Before testing on leaf pigments, we used chromatography to separate the pigment colors in a black marker. The results were actually quite beautiful.

photo 1

In our lab, part of the purpose was to find the Rf value of each pigment. The Rf value stands for Relative Mobility Factor, which is referring to the movement of a substance in relation to the other protein bands. It is calculated by (Dunknown)/(Dsolvent). Dunknown stands for the distance that the solute traveled up the chromatography paper and Dsolvent stands for the distance the solvent traveled up the chromatography paper. Because each substance/pigment has a different Rf value, scientists can study the value in order to identify different types of pigments.

After testing chromatography using marker, we went on the the actual lab: using chromatography to separate the pigments in plant leaves. Our results were as following:

Our group was only able to identify two types of pigments in the green leaf: Carotene (the orange band) and Chlorophyll a (the light green band). In the purple leaf, we were able to identify carotene as well, and some other unknown pigment, which made a purple color. What we noticed is that in the purple leaf, that purple pigment masked the bands of the other pigments – however, it was evident that there were still other pigments there. Our group really enjoyed this lab because it is so aesthetically appealing. However, we found it hard to identify the different colors and to be able to tell where the rings were, because they faded a great deal. In our two green leafs, the results of the Rf value were extremely close (.88 and .89) which correlates with the fact that if the conditions for a set of chromatography are consistent, the Rf value will be constant as well. In plants, pigment is the means of which the energy of the sun is captured for photosynthesis, because pigments absorb the light energy and transfer it, which excites the electrons and thus makes way for the beginning steps of photosynthesis. My major question after doing this lab is how/why plants change color in the fall. I don’t understand how pigments are added or taken away for the start of the new season, or how one suddenly becomes dominant and masks the other.

Thanks for reading! Make sure to MicroSCOPE out the rest of my blog!

*All photos taken by our lab group*

“Affect of Different Colored Lights on Photosynthesis.” Affect of Different Colored Lights on Photosynthesis. CU Boulder, n.d. Web. 04 Oct. 2014.

Light Absorption for Photosynthesis.” Light Absorption for Photosynthesis. N.p., n.d. Web. 04 Oct. 2014.