Fast Week with Progress

The plants that were seeded last week didn’t germinate this week, so the new approach was to use another type of soil known as vermiculite. Vermiculite holds more moisture for less watering for the inexperienced gardener. The green thumb isn’t in my DNA structure, but I still have six pots of regular soil and another six of vermiculite. Also, the Sonora Wheat is still being used but the soy beans were change to Anasazi Beans for germination. Figure 1 below shows the arrangement of seeds planted.

Figure 1. I should start to see some sprouting next week by Monday at least if not then I'm a horrible planter. Furthermore, these seeds grow in places with less water like Arizona or other states along the Southwestern States.

I haven’t sampled this week for my fomite project but I have some potential ideas of where to sample at. The gym has some exercise equipment that can be sampled, and maybe the smartphones students use around campus can be sampled. I can prepare and pour TSA in the contact plates with no problems now.

Furthermore, I took my sample from the residential property across the street from the DB building. Figure 2 and 3 below show how I took my sample with a couple of carpenter tools although the incremental borer would make it easier. Once the sample arrived in the lab, a simple protocol was used to isolate the potential specimen for Agrobacterium. On the other hand, the results were not what I expected because of lack of ascetic techniques and storing areas.

Figure 2. Approaching the area where the sample was taken is shown above. The inside volume is like a pyramid that is scraped out with tools like a flat head screwdriver, mini sludge hammer, box cutter, and some tweezers.

Figure 3.  This side view shows the measured depth of the sample obtained on Wednesday October 16, 2013.

Side Project

The easiest way to perform a natural molecular biological transformation with plants is using bacteria. The bacteria being used is Agrobacterium tumefaciens (A. tumefaciens) that has a plasmid that can be easily transferred to plant cells. Thus, this Gram-negative agent is used as a tool for engineering desired genes into plants. The major cause of plant disease that this bacterium causes is crown gall. Figure 1 and 2 below show some of the pictures I took around off campus sites.

Figure 1. This tree whole trunk is covered with crown gall. The tree is located just across the street from the DB Building at Phoenix College.

Figure 2. The second site of this crown gall is located just a few more feet away near Thomas Rd.

I started preparing my subjects below for germination and the seeds used are soybeans and wheat. Figure 3 and 4 show the pots and location of each per pot.

Figure 3. The seeds used are shown below the pots and the pots are numbered 1-6 just above the rim.

Figure 4. The pots were placed in a 12 hour light incubator for growth around 1 o'clock on October 08, 2013. Note that the typo above in the picture is "seeds" for the "plants"

I hope to identify these bacteria with positive growth of the tumor once introduced to the plant with some type of identification marker like pGLO added to the bacteria. I plan on thinking about how to introduce these bacteria to the growing plants. For example, should I inject the disease causing tumor in the stem, on the leaf, or on the roots? Should I mix with soil or dip plant in solution of A. tumefaciens? Moreover, I look forward to a positive Agrobacterium tumefaciens.

Exciting, But Exhausting This Week

I sampled this week the school text books at the bookstore and library of the campus. The used rental books at the bookstore of Phoenix College were sampled on the cover. The picture below shows the titles of each book sampled below.

 

Figure 1. The numbers on the left side of the picture correspond with the plate sample. Some books were sampled twice on the covers.

Used Rental Books at Bookstore
Plate #	# CFU	Plate #	# CFU	Plate #	# CFU	Plate #	# CFU
1	21	6	10	11	TNTC	16	35
2	4	7	10	12	32	17	TNTC
3	19	8	30	13	6	18	20
4	2	9	15	14	41	19	28
5	11	10	51	15	43	20	41

Figure 2. The books shown above are the reference books used throughout the school year if the student doesn't have enough money for the books in the campus bookstore or elsewhere. They can be checked out for 2 hours maximum if there isn't anyone else waiting for the book.

Reserved Books at Campus Library
Plate #	# CFU	Plate #	# CFU	Plate #	# CFU	Plate #	# CFU
1	5	6	15	11	10	16	9
2	5	7	8	12	1	17	86
3	15	8	42	13	3	18	26
4	2	9	27	14	0	19	15
5	4	10	7	15	18	20	20

The reserved books behind the counter of the campus library were, also, sampled. The picture above show the books used in this experiment.

Although I haven’t had much time in the lab this week, I was pondering if it is possible to insert pGLO into plants for an illumination. Basically, a plant that glows as it grows. For example, there are enzymes that will break open an E.coli cell and combine the pGLO DNA into the ring DNA of the cell for reproduction of glowing bacteria colonies. The process of transferring exogenous DNA information and up taking it into another cell membrane is called Transformation in molecular biology. The video above shows how to conduct animal DNA transformation. The second video below shows how the start of this project might begin with the first steps. Maybe?

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