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Effects of Taxol on Neurospora Crassa Hyphal Tip Growth

Created by Alexandra Fahlgren and James Zabbal

Purpose/Objective

To investigate the effect of Taxol on fungal tip growth in Neurospora Crassa.

Purpose/objective

What is Taxol?

Taxol is a chemotherapy drug.

  • Taxol induces tubulin polymerization and forms extremely stable and nonfunctional microtubules-> leads to cell arrest or cell apoptosis
  • comes from Pacific yew tree

microtubules are essential for rapid hyphal growth

What is Taxol?

Hypothesis

Hypothesis

Hypothesis (Ha): Taxol has an effect on hyphal tip growth

Null Hypothesis (Ho): Taxol has no effect on hyphal tip growth

Protocol

1) Set up microscope for Kohler illumination at 40x magnification

2) Obtain the innoculated Neurospora Crassa agar slides

3) Separate 4 slides into two groups; control and experimental.

Experimental: 50uL growth medium with Taxol

Control: 50 uL growth medium

Both were incubated for 25 miniutes.

4) Place experimental slides under microscope. Use Phase-Contrast (Ph2) and align one end of the ocular scale with a hyphal tip and the other away from the tip.

5) Record the growth of the tip every 2 minutes, over a 16 -minute interval.

3 fungal tips were observed in each group.

Protocol

Results- Control

Results

Figure 1 graph

Figure 1. Average distance travelled by hyphal tip of N. crassa exposed to 50 microlitres of growth medium over a period of 16 minutes. Data shows a linear increase in movement of hyphal tips in control samples with a rate of 0.99 micrometres/minute.

Results- Experimental

Figure 2 Graph

Figure 2. Average distance travelled by hyphal tip of N. crassa exposed to 50 microlitres of taxol over a period of 16 minutes. Data shows a linear increase in hyphal tip growth with a rate of 0.62 micrometres/minute.

T-test

The two-tailed P value equals 0.4102.

By conventional criteria, this difference is considered to be not statistically significant because it is greater than 0.05.

T-test

Questions Answered

How fast do the tips grow?

Control: approximately 0.99 micrometres/minute

Experimental : approximately 0.62 micrometres/minute.

Does each hypha grow at a steady speed or does it vary over time?

Not exactly, but growth increments were fairly similar every 2 minutes over a 16 minute interval.

Do all hyphae grow at the same rate?

No, some hyphae grow faster than others. See experimental data above.

Conclusions

It was predicted that Taxol has an effect on hyphal tip growth in Neurospora Crassa.

  • Despite the t-test, an effect on hyphal tip growth is supported by the data represented in figures 1 and 2. Perhaps the effect is not statistically significant.
  • The experimental slides had a smaller growth rate compared to the control slides.

Therefore...

Therefore Taxol does have an effect on hyphal tip growth. Suggests that microtubules play a role in hyphal tip growth.

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