SALAMANDER SPECIATION Name________________________

The Analysis of Field Observations

BACKGROUND: The small salamanders of the genus Ensatina are strictly terrestrial.
They even lay their eggs on land. Nevertheless, these salamanders need a moist
environment and do not thrive in arid regions. In California, the species Ensatina
eschscholtzii has been studied by R.C. Stebbins at the University of California. This
investigation is based on his work.

PROCEDURE - PART A, COLLECTION AREAS:

1. Imagine that you are working with Stebbins' salamander specimens, some of which
are pictured on the colored sheet provided.

a. In the list below, the salamanders are identified by subspecies (a subspecies is a

geographically restricted population that differs consistently from other populations of
the same species). For example, the first one is Ensatina eschscholtzii croceator,
shortened to E.e. croceator. "croceator" indicates a particular subspecies population of
Ensatina eschscholtzii.

b. The parentheses after each subspecies name contain a number and a color. The
number is the total of individuals Stebbins had available for his study. The color is the
one you should use for that subspecies when you plot its collection area on the
California map.

c. Following the parentheses is a list of grid codes indicating where (on the map grid)
the subspecies was collected. For example, 32/R means that one or more specimens
were collected near the intersection of horizontal Line 32 and vertical Line R.

d. The letter before the subspecies name indicates the corresponding salamander
picture on the color sheet. For example, E.e. eschscholtzii is picture b on the color
sheet.

2. Plot each collection area by filling in the corresponding square on the California
map grid. Color in the square above and to the left of the point where the specified
grid coordinates cross. For example, the square in the upper left-hand corner is 1/A.
Use the colors indicated for each subspecies population (listed below) to make a
distribution map of Ensatina eschscholtzii in California.

a. E.e. croceator (15; brown): 32/R, 32/S, 30/T, 31/T

b. E.e. eschscholtzii (203; red): 30/M, 32/O, 34/S, 35/V, 36/W, 35/Z, 38/Y, 40/Z
c. E.e. klauberi (48; blue): 36/Z, 38/a, 39/a, 40/a
d. E.e. oregonensis (373; purple): 9/B, 7/E, 6/E, 13/C, 10/C, 7/D, 15/D
e. E.e. picta (230; yellow): 2/B, 2/C, 3/C, 4/C
f. E.e. platensis (120; green): 8/J, 10/J, 11/M, 13/M, 15/M, 15/O, 17/M, 15/P, 20/Q,
24/S, 21/R, 25/T, 26/U
g. E.e. xanthoptica (271; orange): 17/G, 17/F, 19/H, 19/O, 20/I, 20/J, 21/I

3. Answer the Discussion Questions for Part A, based on your work and
interpretations, and your discussions with partners.

3a. Is the species uniformly distributed? Use your knowledge of the species' ecological
requirements (see "Background") to offer an explanation of its distribution. Are there
any other factors that might affect distribution?

3b. Consider the physiography of California. Does the species seem more
characteristic of mountain areas, or of valley areas?

3c. Do you expect any pattern in the distribution of subspecies? Why or why not?

3d. Examine the salamanders on the color sheet. Note that some subspecies have
yellow or orange spots and bands on black bodies. Some have fairly plain, brown-
orange bodies. One has small orange spots on a black background. There are other
differences as well: for example, some of them have white feet. Now refer to your
distribution map. Does there appear to be any order to the way these color patterns
occur in California? For example, do the spotted forms occur only along the coast? Do
spotted forms occur in the north and unspotted ones in the south? What do YOU find?

3e. Subspecies E.e. eschscholtzii (specimen b) and E.e. klauberi (specimen c) are
different from each other. What relationship is there between their distributions?

PROCEDURE - PART B, COLLECTION AREAS:

1. You also may wonder if there might be additional subspecies for which you have no
specimens. A biologist faced with this question would leave the laboratory and go into
the field to collect more specimens. Imagine that you have done this and returned
with the following data:

h. Unidentified population 8 (44; pink): 4/I, 5/H, 7/H, 7/F, 6/J, 9/F
i. Unidentified population 9 (13; black): 28/T, 27/T, 26/T, 28/S, 29/T
Mark with crosshatch the following places that were searched for Ensatina
without success: 11/I, 14/I, 17/K, 22/N, 26/Q, 5/M, 32/U, 32/a, 35/f
2. Answer the Discussion Questions for Part B, based on your work and
interpretations, and your discussions with partners.

2a. According to Stebbins, the unidentified populations are not additional subspecies.
What, then, is the probable genetic relationship of populations 8 and 9 to the
subspecies plotted on the map?

2b. Why is it unlikely that you would ever find individuals combining characteristics of
E.e. picta and E.e. xanthoptica ?

2c. Explain why you think Stebbins concluded that there is only one species of
Ensatina in California.

2d. Suppose that volcanic activity in northern California should become violent and
completely destroy all the salamanders in that region. How would this event
affect the species Ensatina ?

2e. What do you think would account for the one record of E.e. xanthoptica in the
Sierra, with the rest of that subspecies occurring along the coast?
(Advanced) PROCEDURE - PART B, COLLECTION AREAS:
1. You may wonder if there are salamanders in some areas for which you have no
records. You also may wonder if there might be additional subspecies for which you
have no specimens. A biologist faced with these questions would leave the laboratory
and go into the field to collect more specimens. Imagine that you have done this and
returned with the following data:

b. E.e. eschscholtzii (16; red): 36/Z, 41/Z, 33/M, 34/W, 34/U

c. E.e. klauberi (23; blue): 40/b, 40/Z, 36/a
h. Unidentified population 8 (44; pink): 4/I, 5/H, 7/H, 7/F, 6/J, 9/F
i. Unidentified population 9 (13; burgundy): 28/T, 27/T, 26/T, 28/S, 29/T
k. Unidentified population 11 (131; turquoise): 23/J, 24/K, 24/I, 29/M, 25/J, 25/I
l. Unidentified population 12 (31; black): 6/C, 7/C, 6/B

2. Mark with the color gold the following places that were searched for Ensatina
without success: 11/I, 14/I, 17/K, 22/N, 26/Q, 5/M, 32/U, 32/a, 35/f

Specimens of populations 8 (specimen h) and 9 (specimen i) are shown in the

color pictures. (There are no illustrations for populations 11 and 12).

3. Answer the Discussion Questions for Part B, based on your work and
interpretations, and your discussions with partners.

1. According to Stebbins, the unidentified populations are not additional subspecies.

What, then, is the probable genetic relationship of populations 8, 9, and 11 to the
subspecies plotted on the map?

2. On this basis, describe (or make a colored drawing of) the appearance you would
expect specimens of population 11 to have.

3. Why is it unlikely that you would ever find individuals combining characteristics of
E.e. picta and E.e. xanthoptica ?

4. Look at the distribution of the original collections of E.e. eschscholtzii and E.e.
klauberi. What reasons were there for trying to collect additional specimens from
extreme southwestern California?

5. How do the results of the additional collections differ from the results in other
places where two different populations approach each other?
6. Bear in mind the biological definition of a species and also the appearance and
distribution of the named populations of Ensatina. Which one of these populations
could be considered a species separate from E.e. eschscholtzii ? (The population was
indeed once considered by biologists to be a separate species).

7. Now imagine that, while examining salamanders in another collection, you find
specimen j from population 10, shown in the colored photo. Compare its
characteristics, especially the spotting pattern, with those of the named populations.
Also, consider the distribution of these populations. Between which two populations is
this specimen most likely a hybrid? On your map, draw a line where you might expect
to collect other specimens like this one.

8. In a brief paragraph, explain why you think Stebbins concluded that there is only
one species of Ensatina in California.

9. Suppose that volcanic activity in northern California should become violent and
completely destroy all the salamanders in that region. How would this event
affect the species Ensatina ?

10. What do you think would account for the one record of E.e. xanthoptica in the
Sierra, with the rest of that subspecies occurring along the coast?

For teachers: Answers to Discussion Questions

Discussion for Part A:
3a. No. In those areas where salamanders do not occur, there probably are specific limiting factors, such as
arid or semiarid conditions. Historical factors, such as human habitation, also can affect the distribution.

3b. Salamanders occur in the mountainous regions, except in mountainous desert ranges.

3c. The fact that subspecies are geographical variations within a species suggests there is an order to the
distribution of these sub-species. Because of ancestral relationships, adjacent species should be more like
each other than widely separated subspecies.

3d. The spotted forms tend to be in the inland Sierra Nevada and the unspotted forms along the coast, except
in southern California. where spotted and unspotted forms occur together.

3e. Populations of E e. eschscholtzii and E e. klauberi occur in the same area in southwestern California.
Discussion for Part B:
2a. They represent genetic intergrades (hybrids) between subspecies.

2. The drawing should be an intergrade between E. e. eschscholtzii and E. e. xanthopica. Making such a
drawing is a difficult but challenging assignment, requiring a bit of imagination. There are clines from north
to south between the two adjacent subspecies (E. e. xanthopica in the north and E e. eschscholtzii in the
south). In general, from north to south the eyelids tend to become lighter and the dark pigmentation of body
tends to disappear or become restricted to dots. To be more specific, toward the south:
a. Lemon-yellow eye patch disappears.
b. Orange coloration of ventral surface becomes restricted to underside of limbs and tail or is lost (sketches
will not show this, of course).
c. Yellow dots on sides of back appear.
d. Small black dots on sides of back appear.
e. Tip of tail becomes lighter.

2b. These two are geographically isolated by several hundred miles, with another subspecies occurring
between them.

2c. The map shows intergrades between all the subspecies except the two that exist together in southern
California. It would seem likely that subspeciation has taken place from a common ancestor in the north
(closely related to E. e. picta), down the two separated mountain chains. The two subspecies that exist
together without intergrading must have become so different from each other that they are reproductively
isolated. You can see why these two were thought to be separate species before all the information on the
intergrades was available.

2d. It would break the chain of interbreeding subspecies because E. e. eschscholtzii and E. e. klauberi at the
southern end of the range do not interbreed. This would produce two reproductively isolated populations—
two species.

2e. There are several possible explanations. The specimen may have been introduced accidentally by
humans, or irrigation in this region may have allowed the specimen to move across the broad valley. More
intensive collecting in the valiey might show more specimens in the inland mountains, with perhaps a
connection between the coastal and inland populations in the not too distant past. In actuality, more than just
this one individual were located in the inland mountains, in thk same general region.