Saturday, September 7, 2024

UNDERSTANDING GENOTYPE AND PHENOTYPE

 

Name:    UNDERSTANDING GENOTYPE AND PHENOTYPE

Activity:                Students will get a bag of shells from two beaches, sort them, identify them and look at variations.

Level:                    3

Developer:         Alberto Mimo                                                                                                   

Site:                       Classroom or Laboratory with access to a computer and excel.

Subjects:             Genetics, statistical analyses.

 

Introduction

Darwin and Russell in the 1800’s were interested in understanding how new species came about. They concluded that somehow random changes in their phenotype were best adapted to an environment or situation and those organisms that did not fit would die out, or lived at a disadvantage because they were not well fitted to the new condition. This is in essence the idea of evolution based on natural selection due to the survival of the best-fitted animal or plant. At the same time, a man named Jean-Baptiste Lamarck theorized that animals evolved by the force of adapting themselves to a new environment and changing their phenotype. Long-neck giraffes get longer necks by trying to reach the leaves on the taller trees. In this lesson, we explore the idea of phenotypic plasticity based on environmental conditions or contact with other species

A living organism’s DNA contains the information that will be transferred to its progeny. Some information is not shown physically or physiologically, but it is all there in their genome. Some of the information is displayed in every organism either in its shape or form or in its physiology. (Read about Gregor Mender,  1822-1884) One is called the organism’s genotype and the other is the organism's phenotype.

Organisms within a species are not always exactly alike, there is a variation within each species that refers back to the exact genome they have. In people, some kids have blue eyes and some dark brown, but this variability has to be all accounted for unless there has been a mutation. Mutation could lead to new species, but that is only when that mutation shows to provide that organism with an advantage. See Darwin and Russel's theory of Evolution.

Variability within one species could have been led by small changes in the habitat, other species' competition, or a predator/ prey relationship. (Anurag A. Agrawal in his paper “Phenotypic Plasticity in the Interactions and Evolution of Species”) Published by the Journal of Ecology in 2001 shows how these changes can be derived from competition, mutualism, predation risk, and parasitism/herbivore or food quality.

Thomas J. DeWitt, Andrew Sih, and David Sloan Wilson in their paper “Cost and limits of phenotypic plasticity” Trends in Ecology & Evolution, 1998, discuss the idea that variations on an organism's phenotype have limits and cost. Some of this cost includes Maintenance and Production of the Phenotypic Change, Information Acquisition Cost, Development Instability, and Genetic Cost, but at the same time, there is a benefit of survival.

Some of the terminologies associated with phenotypic plasticity are: (Phenotypic Plasticity and the origin of diversity. Mary Jane West-Eberhand, Annu. Rev. Ecol. Sust. 1989 20:249-78).

  • Alternative phenotypes: Two or more forms of behavior, physiological response, or structure maintained in the same life stage in a single population and not simultaneously expressed in the individual.
  • Conditional (or condition – sensitivity): The alternative adopted by a particular individual or at a particular time depends on environmental conditions.
  • Allelic-Switch: The alternative adopted by a particular individual depends on the allele(s) present at one or more genetic switch loci.
  • Combine switch: The alternative adopted depends on a combination of allelic and environmental factors.
  • Polyphenism: The existence of environmental-cued alternative phenotypes in a population.
  • Polymorphism: The existence of morphological district alternatives in a population (usually: “allelic-switch alternatives”.

 

Hypothesis

Slipper Shells live at the bottom of the Long Island Sound.  They form a tight community of shells at the bottom of the seawater and are constantly subject to being swept away by the current caused by the incoming and outgoing tides. In areas with narrow channels the effort by the shells to stay in place is greater and in more level areas where the current is less strong the effort of the shells is also less.  The speed of the water at different locations within the Sound changes depending on the site, for example, areas located at the entrance of the sound called the race have stronger currents.

The hypothesis here is that at locations where the speed of the water current is high; shells will transform their shape to be more flat reducing the friction with the water. Shells that live in more tranquil areas will be shaped more bumped or higher in the top. So we have two possible different phenotype shapes on these shells. The idea here is to take measurements and see if we can find differences within collection sites. Have C. fornicata shells a phenotypic plasticity?

Read Phenotypic Plasticity and the Origen Of Diversity. Mary Jane West-Eberhard. Annu. Rev. Ecol . Syst. 1989. 20:249-78

Natural History and Anatomy of the Slipper Shell (Crepidula fornicata L.)

Slipper Shells live out in the bottom of the Long Island Sound. They are found in amazing quantities and compete for habitat with oysters and other organisms. These mollusks stack one on top of the other forming a thick layer. During reproduction, they form clusters where the ones at the bottom are females and the ones on top are males. The eggs are found in sacks that are fertilized in the water and released. These organisms are hermaphroditic, so they change sex as they grow. Starting as males and growing into a female, they live for about 7 to 10 years. You can count the number of growth rings. When they die they are carried away by the tide and the water current. They have very few predators so they are a nightmare to the oyster industry.

A cluster of Slipper Shells from the bottom of the LIS (Long Island Sound) mixed with mud from the site. These shells were collected using a drudge from a science vessel. (Next 3 pictures)

                  

 

Mud and shells covers the see floor

 

Some are on top of other; the lower one is a female


See diagram for their anatomy.

 

Anatomy of the slipper shell


Slipper shells do not move much, they crawl at the beginning but once they find the perfect spot they attach permanently. Their typical food is plankton and other particulate organic matter. Their growth rate is between 0.04 to 1.11 mm/day.

Their abundance can be up to 4,770 individuals per m2. They prefer a substrate made up of small boulders, and muddy gravel and they live at the sublittoral fringe where they are sheltered. Their preference is a salinity of 18 to 40 milligrams per centiliter and a water flow of 1 to 3 knots.

Sites for your Collection

You can collect anywhere there are slipper shells but I have included three sites that are located somewhere south of the state, Coastal Center at Milford Pont, a north site at Hammonassett Beach State Park in Madison, and one additional site at the Race of the Long Island Sound at Watch Hill Cove in Rhode Island. It would be best to do this research on-site without taking the shells to school. These slipper shells form the natural habitat of the area.

 

Sites for your Collection

You can collect anywhere there are slipper shells but I have included three sites that are located somewhere south of the state, Coastal Center at Milford Pont, a north site at Hammonassett Beach State Park in Madison and one additional site at the race of the Long Island Sound at Watch Hill Cove in Rhode Island. It would be best to do this research on site without taking the shells to school. These slipper shells form the natural habitat of the area.

Site One


Costal Center at Milford Point

 

Site two


Hammonasset Beach State Park Meigs point

 

 

Site three


Watch Hill Cove, in Rhode Island

Where do the dead shells come from in your collection?

Where do the dead shells come from in your collection?

Before you start your collection you will need to acquire a Scientific Collecting Permit for marine organisms from the CT Department of Energy and Environmental Protection. You can get the forms at their website (https://portal.ct.gov/DEEP). If you intend to take all the measurements at the beach you do not need to get a permit. The scientific collecting permit will cost you $25 and you have to fill out the correct forms.

The shells that you will collect are found on the beach. These are dead shells that were living in the water near your location that for some reason died and washed into the beach by the tides or storms. They are the perfect habitat for Piping Plovers

You need to collect the shells when it is low tide because we also want you to collect a cluster of live shells deposited in the intertidal area by the tide. There should be between 3 and 7 shells that you will use to learn more about the anatomy of the slipper shells. The other slipper shells you are collecting are just shells without the actual animal on them. Just walk close to the water's edge and look for a cluster. There will be several, take only one and throw the others into the water.

Place the cluster into a jar with 70 % alcohol and label it.

 

 

Place the cluster into a jar with 70 % alcohol and label it.

Species: Crepidula fornicata L.

Date:

Location:

Your school name:

Inventorying the beach

Based on the current literature, there are 3 different slipper shells in the Northeast; Crepidula convexa, convexa which is brown with striped or dots of reddish brown, up to 1.3 cm. long, and is less common than other slipper shells; attaches to shells and rock and other hard objects.  There is the Crepidula plana, also known as eastern white slipper shell, pearly white up to 3.3 cm. long often found attached to the underside of horseshoe crab shells and inside large snail shells occupied by hermit crabs. And finally, there is Crepidula fornicata.

Shells accumulated on the beach


Shells accumulated on the beach


Close up of shells

 

Shells clearly left by the tide


Variety of shells on the beach

 

In addition to collecting the slipper shells, we also need to complete an inventory of other organisms found in the same area. A sandy beach about 15 ft. from the high tide line and about 75 feet in linear length parallel to the beach. Remember that many small invertebrates live buried in the sand. Bring a small shovel.

To do this just get several small buckets or plastic bags, one per every two students, and collect just one good specimen, in good shape of every species you see. Sit by the side of the beach and identify the organisms. Make a list. Then take a picture and let them go.

Use the following list to write the length and height of two sets of shells of 45 individuals. Count the rings of 10 shells and each ring is one year. Write the information on the size of this list.

 

To do this just get a number of small buckets or plastic bags, one per every two students and collect just one good specimen, in good shape of every species you see. Sit by the side of the beach and identify the organisms. Make a list. Then take a picture and let them go.

 

Shell number

Age

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


 

Length and Height measured in mm.

 


Length

Height

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 



 

Measuring each shell

Measure each shell in millimeters using calipers.

 

Length: Largest length of the shell.

 

 

 

 

 

 

 

Height: Place the calipers touching both sides of the shell and the top of the shell.



 

Doing the analyses

Doing the analyses

To show you what kinds of things you can do, here is an example:

  1. I went to West Haven to a small beach and collected shells then I went to Hammonnaset State Park and did the same thing, counted them, measured them, and calculated the ratio of height to length. These are the measurements of the shells.

2.        


Shells A West Haven


Lenth mm

Height mm

Ratio

3788

890

0.23

2556

965

0.38

2757

1052

0.38

2646

1080

0.41

2563

1084

0.42

2418

1110

0.46

2754

1119

0.41

3028

1157

0.38

2927

1179

0.40

3075

1186

0.39

3108

1192

0.38

2927

1200

0.41

2941

1211

0.41

2896

1226

0.42

3004

1228

0.41

2668

1247

0.47

3213

1249

0.39

2786

1257

0.45

2750

1259

0.46

3001

1275

0.42

2952

1278

0.43

2796

1280

0.46

2436

1280

0.53

3307

1297

0.39

2807

1298

0.46

3031

1306

0.43

3398

1307

0.38

3022

1319

0.44

2809

1327

0.47

2774

1336

0.48

2806

1337

0.48

3089

1355

0.44

2908

1368

0.47

3010

1390

0.46

3170

1398

0.44

3100

1420

0.46

3288

1447

0.44

3395

1450

0.43

3231

1490

0.46

3439

1500

0.44

3715

1527

0.41

3425

1532

0.45

3300

1554

0.47

3164

1778

0.56

3625

1791

0.49

 

 


Shells B Hammonasset


Length mm

Height mm

Ration

2266

616

0.27

2673

682

0.26

2556

952

0.37

2591

958

0.37

2521

982

0.39

2660

990

0.37

3289

991

0.30

3226

1010

0.31

2860

1057

0.37

2585

1084

0.42

2770

1103

0.40

2693

1108

0.41

3153

1110

0.35

2802

1117

0.40

3170

1141

0.36

2985

1143

0.38

2714

1151

0.42

2634

1156

0.44

2890

1169

0.40

2814

1177

0.42

2984

1202

0.40

3226

1208

0.37

2693

1211

0.45

2855

1211

0.42

2971

1245

0.42

2916

1272

0.44

3036

1287

0.42

2953

1288

0.44

3112

1290

0.41

2971

1296

0.44

3288

1306

0.40

2986

1307

0.44

2952

1327

0.45

3296

1328

0.40

3202

1345

0.42

3446

1353

0.39

3033

1363

0.45

3482

1374

0.39

3161

1388

0.44

3353

1397

0.42

3171

1453

0.46

3249

1476

0.45

2984

1512

0.51

3454

1569

0.45

2600

1651

0.64

 

3.       Next I organized them according to the height, from smaller to larger. This is how you do this is in excel:

4.       Go to data


5.       Select sort:


Click OK:

And you are done; all the numbers are lined and sorted by height from lower to largest

6.       Now you can make a line graph showing the progression of height on all these shells.

 

As you can see there are not too many shells that are flat in either sample. If we take the next sample we get:


No flat shells either

Using statistics to compare numbers

We will compare the height of the shells. If the two sets of shells are similar in height then there is not much difference between the two populations. If the shells height is significantly different then there is a difference between the two populations. To do this we will use the t- test in statistics. The t -test is simple to use and we will use the null hypothesis to learn from the results.  The null hypothesis says that if our t value (we will calculate that using excel) is higher than the t Critical Value a .05 percent difference then we do reject the null hypothesis and determine that the two sets of heights are different and come from the different populations, but if the t value is lower  than .0 5 percent then we do not reject the null hypothesis and determine that the two sets are from the same population the same and are comparable.

To do this is in excel do the following:

1.       Line up the two samples of a blank sheet; use the height of the shells

A

B

890

616

965

982

1052

952

1080

1084

1084

958

1110

1651

1119

1156

1157

990

1179

682

1186

1108

1192

1211

1200

1151

1211

1103

1226

1117

1228

1177

1247

1211

1249

1057

1257

1169

1259

1272

1275

1327

1278

1288

1280

1245

1280

1296

1297

1202

1298

1512

1306

1143

1307

1307

1319

1363

1327

1287

1336

1290

1337

1110

1355

1388

1368

1141

1390

1453

1398

1345

1420

1208

1447

1010

1450

1476

1490

1306

1500

991

1527

1328

1532

1397

1554

1353

1778

1569

1791

1374



2.       To use excel  go to data, look at the far right hand corner and choose data analyses.

3.       This will come up


4.       Choose t-Test paired, hit OK

5.       Fill the data for the two shell heights and it will automatically do all the calculations.

6.       Press OK to the next pop-up


 

7.       These are the results for the t-test

Mean

1300.689

1207.911

Variance

32003.49

41482.49

Observations

45

45

Pearson Correlation

0.568208


Hypothesized Mean Difference

0


df

44


t Stat

3.474853


P(T<=t) one-tail

0.00058


t Critical one-tail

1.68023


P(T<=t) two-tail

0.001161


t Critical two-tail

2.015368

 

 

The t value is 3.47 and the critical value is 2.01 Therefore the t value is larger than the critical value and we reject the null hypothesis and conclude that there is a difference between the two sets of numbers of shells heights. Maybe there is some Lamarckism on the development of the shells. Their Phenotype may be malleable.

Part two of analyses

 

We took two measurements from each shell: The length and the height. Using these two numbers we calculated a ration, by dividing the height by the length. This is a number between 0 and 1. If the height was the same as the length then the answer would be 1.0.  Any height smaller that length is a decimal point number. The lower the height the larger the answer up to 1.0.

Let’s compare the list of ratios and see what we get for a t-test value.

Shells A   Shells B

A

B

0.23

0.27

0.38

0.26

0.38

0.37

0.41

0.37

0.42

0.39

0.46

0.37

0.41

0.3

0.38

0.31

0.4

0.37

0.39

0.42

0.38

0.4

0.41

0.41

0.41

0.35

0.42

0.4

0.41

0.36

0.47

0.38

0.39

0.42

0.45

0.44

0.46

0.4

0.42

0.42

0.43

0.4

0.46

0.37

0.53

0.45

0.39

0.42

0.46

0.42

0.43

0.44

0.38

0.42

0.44

0.44

0.47

0.41

0.48

0.44

0.48

0.4

0.44

0.44

0.47

0.45

0.46

0.4

0.44

0.42

0.46

0.39

0.44

0.45

0.43

0.39

0.46

0.44

0.44

0.42

0.41

0.46

0.45

0.45

0.47

0.51

0.56

0.45

0.49

0.64

 

Results for the t- Test

t-Test: Paired Two Sample for Means




 

Variable 1

Variable 2

Mean

0.432222

0.407333

Variance

0.002504

0.003643

Observations

45

45

Pearson Correlation

0.570153


Hypothesized Mean Difference

0


df

44


t Stat

3.21146


P(T<=t) one-tail

0.001235


t Critical one-tail

1.68023


P(T<=t) two-tail

0.00247


t Critical two-tail

2.015368

 

 

 

T=Value =3.21

Crit -t = 2.01



T value is > Critical  -t

Reject the Null Hypothesis

 

By using the t-test on both calculations we get the same results. The shells from site A are not the same as from site B therefore each individual slipper shell grows at different rates.  In cases such as these we should continue the research by setting up an experiment that includes a control group. Additional information could help to answer some questions such a where the shells come from and what is the water current at that site.

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