A4.

12S

Activity 4.12 Natterjack toads and

genetic diversity

Student

Purpose
To consider how genetic diversity can be measured
To calculate and compare the heterozygosity index for two populations
Heterozygosity index
The natterjack toad (Bufo calamita) is Britains rarest amphibian. It is very restricted in
distribution, due to loss of its preferred habitat of heath or sand dunes. In Britain there are
now very few populations, with a stronghold around Formby and Ainsdale on Merseyside.
Trevor Beebee and his research group at the University of Sussex have investigated the
genetic diversity of natterjack toad populations and attempted to correlate this diversity with
the genetic fitness of the animals.
The genetic fitness of a population reflects the average fitness (the likelihood of survival
and successful breeding) of the individuals within it. Measurable quantities such as rate of
growth, resistance to disease and spawn size will all affect genetic fitness.
Genetic diversity has been measured by preparing DNA fingerprints for individuals within
the populations. (You will discover exactly how these fingerprints are made in Topic 6.) By
examining the prepared DNA fingerprints it is possible to calculate how many of the
different gene loci are heterozygous, i.e. have more than one allele present. The proportion of
genes which are present in heterozygous form can be expressed as a number called the
heterozygosity index.
Figure 1 represents the DNA fingerprint of 10 DNA microsatellite sequences for one
individual. Note that sequences I, VI and VII give only a single band, indicating that the
individual is homozygous for these sequences. The other seven sequences each have two
bands showing that the individual is heterozygous for each of these sequences.

Figure 1 The banding pattern for 10 DNA microsatellite sequences in an individual.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original.

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A4.12S

Student

Activity 4.12 Natterjack toads and genetic diversity

A heterozygosity index for each sequence can be calculated using the equation:
number of heterozygotes

heterozygosity index =

number of individuals in the population

The DNA fingerprint banding patterns for sequence VII in a population of 21 individuals is
shown in Figure 2. The heterozygosity index for this sequence is:
heterozygosity index (VII) = 7/21 = 0.333
An average heterozygosity index for the population is determined by taking the mean of all
of the heterozygosity index values for each of the microsatellite sequences studied in the
individuals of this population.
The heterozygosity index is a useful measure of genetic diversity. Because it is a proportion
rather than an absolute number, it is unaffected by the sample size.

Figure 2 Fingerprints for microsatellite VII in 21 individuals.

The research group let by Trevor Beebee obtained the results in Table 1 below.
Table 1

Population number

Population size/number of
clumps of spawn laid

33

11

146

17

87

20

Tadpole growth rate/mm per

week

5.0

2.8

4.6

4.1

5.6

5.0

Average heterozygosity

0.269

0.189

0.293

0.257

0.336

0.325

Questions
Q1 Plot tadpole growth rate against average heterozygosity. You could use an automated
programme such as Microsoft Excel to do this, or you can plot the data by hand.
Q2 What do these results suggest about the benefits for genetic diversity (heterozygosity) in terms
of genetic fitness? Use results from the table and from your graph to support your conclusions.
Q3 Two other small populations of toads were studied. DNA fingerprints were prepared and are
shown in Figure 3. Use these DNA fingerprints to calculate the heterozygosity index for these
two populations.
Q4 Use your results to predict the approximate tadpole growth rate for members of these
populations. Which of the two populations is more likely to succeed over a lengthy period of
time? Suggest three environmental factors which should be controlled if these growth rate
results are to be compared in a valid way.
If you are interested in natterjack toad conservation, why not conduct an Internet search for
information on these animals? You will find more data on their distribution and on conservation
measures being taken to protect them.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original.

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A4.12S

Activity 4.12 Natterjack toads and genetic diversity

Student

Figure 3 DNA fingerprints for microsatellites in two toad populations.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original.

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