lab walkthrough

Laboratory Practical Overview

• The purpose of the laboratory practical is to conduct an experiment regarding the enzymatic latch using peat collected from the blanket bog (midnight).

Enzymatic Latch in Peatlands

• Concept of the Enzymatic Latch:

  • It refers to the suppression of organic matter breakdown in peatlands, leading to the accumulation of semi-decomposed organic matter (peat).

  • Phenolics:

  • Chemicals found in plants that can suppress microbial activity, particularly by inhibiting extracellular enzyme activity.

  • Released by mosses (e.g., sphagnum moss) as trans-sphagnum acids.

Hypothesis

• The hypothesis states that phenolic compounds can strengthen the enzymatic latch:

  • Proposed Outcome: Addition of phenolic compounds to peat will lead to a more robust suppression of decomposition.

  • This is hypothesized to be due to the inhibition of hydrolase enzyme activities, which are key for decomposition.

Hydrolase Enzymes

• Role of Hydrolases:

  • They are essential enzymes that facilitate the breakdown of organic matter.

  • Increased presence of phenolics is expected to prevent these enzymes from functioning effectively, leading to reduced decomposition rates.

Implications of Reduced Decomposition

• Decreased decomposition results in lower carbon release:

  • Less breakdown of organic matter means less mineralization of carbon into forms such as CO2 or methane.

  • Potential for carbon to accumulate in peatlands, further enhancing their role as carbon sinks.

Experimental Procedure

1. Peat Collection and Preparation:

   • Collect peat samples from the midnight bog.

   • Homogenize the peat to ensure uniformity.

   • Prepare 10 gram samples of peat for experimentation.

2. Setting Up Experiment:

   • Prepare eight 50 ml centrifuge tubes:

     • For Controls:

     • Add 10 g of peat and 10 ml of distilled water to four tubes.

     • For Treatments:

     • Add 10 g of peat and 10 ml of phenolic-rich solution (phenolic powder mixed with distilled water) to another four tubes.

   • Create a slurry for both controls and treatments and allow to sit for about one hour.

3. Testing Enzyme Activities:

   • After one hour, take out one gram samples from each tube to analyze hydrolase enzyme activity.

   • Perform enzyme assays for each sample to quantify enzyme activities.

4. Data Analysis:

   • Compare enzyme activities between treatments (phenolic-rich samples) and controls (water samples).

   • Expected Results: Enzyme activities in the phenolic treatment are anticipated to be lower than those in the control, indicating less decomposition and therefore less CO2 release.

Conclusion and Expectations

• If the hypothesis holds true, findings will indicate that higher phenolic content leads to more effective suppression of decomposition, allowing for greater carbon retention in peatlands, ultimately enhancing their role as carbon sinks.

• Students are encouraged to review protocol videos for experimental methods and detailed procedures.