DRAFT: This module has unpublished changes.

Phytochrome Laboratory

 

This experiment will take place over the course of five weeks, during which you will expose Wisconsin Fast Plants (Brassica rapa) to different light regimes and then detect the plants responses through both careful phenotype observations and changes in phytochrome A and B protein levels.

 

Outline:

Week 1: Setup

Plant eight Fast Plants (two of each kind) in a random order and decide on an experiment involving how much of each kind of light you will expose your plants to.

  • Our group (Group 5) chose to set our LED lights to Far Red (9), Green (1), Blue (1), and Red (0).

Each day, at 4pm, one member from each group must arrive to measure both stem height and leaf area for each plant for five days. Day 1 will be the basis for comparison to determine percent growth per day and leaf area per day.

 Week 2: The Biochemistry begins

This week you will extract total protein from your samples and quantify how much you have. You will also run an SDS-PAGE gel.

Week 3: Blotting 

This week you will conduct a Western Blot. This will involve transferring your proteins to a membrane, crosslinking them and then washing them with antibody.

Week 4: Development

This week you will develop your Western Blots. Take images and quantify the amounts of the two proteins that you are looking at. You will then try to tie these data into your phenotypic observations that you have collected and analyzed previously.

Week 5: Presentation

 

Results

Phenotype:

Figure 1. Average percent plant growth per day by plant type. (R2 = 0.009995, DF = 3, F = 0.067, P = 0.976, q = 2.799, Tukey: All A)

 

Figure 2. Average leaf area by plant type per day. (R2 = 0.040, DF = 3, F = 0.2786, P = 0.8402, q = 2.799, Tukey: All A).

 

DRAFT: This module has unpublished changes.