Guide to STPM Practicals For Third Term

Construction of a Dichotomous Key Using Local Specimens

This experiment enables students to: 1. Identify the external features (morphology) of an organism to be grouped into a phylum or a class 2. Increase their knowledge in the field of taxonomy 3. To use a dichotomous key to identify unknown plants or animals. Organisms such as Amoeba sp. (slide), Hydra sp. (slide), ant, snail (garden snail), shrimp, Marchantia sp. (moss), Dryopteris sp. (with sori on the undersides of the fronds) and grass were examined under the microscope or using a magnifying glass ( 10). The opposing features, which clearly differentiate the organisms from one another, were noted down. A dichotomous key for this group of organisms was then constructed up to the category of phylum and class. Characteristics of organisms noted: Organism Grass Dryopteris sp. Characteristics Green, containing chlorophyll; does not produce spores; has flowers, roots, stems and leaves Containing chlorophyll; produces spores; fronds sub-divided into pinnae and pinnules bearing sori on undersides

Marchantia sp. Containing chlorophyll; produces spores; thalloid body with rhizoids; has gemma cups, antheridiophores and archegoniophores as reproductive structures Amoeba sp. Hydra sp. Organism without chlorophyll; unicellular. Multicellular; soft cylindrical body without exoskeleton; body wall comprises of two layers of cells ectoderm and endoderm; has tentacles, a gut cavity and a mouth Multicellular; with exoskeleton; has soft muscular foot and calcareous shell Invertebrate with jointed legs; body divided into head, thorax and abdomen; has three pairs of legs; wingless Invertebrate; body divided into cephalothorax and abdomen; has two pairs of antennae and five pairs of jointed legs

Snail Ant Shrimp

From the above observations, it can be inferred that, Marchantia sp. is able to live successfully in highlands and damp areas due to its special rhizoids. The structural differences between Marchantia and Dryopteris are that Marchantia has a thalloid body and gemmae cups whereas Dryopteris has roots, stems, and leaves with the presence of sori on the underside of its fronds. Ant and shrimp share the same phylum, Arthropoda as both have segmented body with exoskeleton and jointed legs. The structural differences between them can be seen in the table below: Ant Shrimp

(a) Segmented body with exoskeleton which is Segmented body with exoskeleton which is divided into head, thorax and abdomen divided into cephalothorax and abdomen (b) 3 pairs of jointed legs (c) A pair of antennae (d) Tracheal system for respiration 5 pairs of jointed legs 2 pairs of antennae Gills for respiration
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A simple spider key, showing contrasting characteristics can be used to differentiate the organisms for the purpose of constructing a dichotomous key.
Organisms Organisms with chlorophyll Marchantia, Dryopteris, grass non-flowering/producing spores flowering/not producing spores unicellular multicellular Hydra, ant, snail, shrimp Marchantia, Dryopteria Without archegonia and antheridiophores Dryopteris Grass with archegonia and antheridiophores Marchantia Amoeba without tentacles with tentacles

Organisms without chlorophyll Amoeba, Hydra, ant, snail, shrimp

Ant, snail, shrimp with leg Ant, shrimp without leg Snail

Hydra

six legs

ten legs

Ant

Shrimp

Dichotomous key A1 With chlorophyll A2 Without chlorophyll B1 Does not produce spores but has flowers, roots, stems and leaves B2 Produces spores C1 Fronds sub-divided into pinnae and pinnules bearing sori on undersides C2 Thalloid body, has gemma cups, antheridiophores and archegoniophores as reproductive structures D1 Unicellular D2 Multicellular E1 Soft cylindrical body without exoskeleton. Body wall comprises of ectoderm and endoderm. Tentacles, gut cavity and mouth present E2 With exoskeleton F1 Invertebrate with soft muscular foot and calcareous shell F2 Invertebrate with jointed legs G1 Invertebrate; body divided into head, thorax and abdomen. Has three pairs of legs; wingless G2 Invertebrate; body divided into cephalothorax and abdomen. Has two pairs of antennae and five pairs of legs

refer to B refer to D Phylum Angiospermae (Grass) refer to C Phylum Filicinophyta (Dryopteris) Phylum Bryophyta (Marchantia) Phylum Protoctista (Amoeba) refer to E Phylum Cnidaria (Hydra) refer to F Class Mollusca (Snail) refer to G Class Insecta (Ant) Class Crustacea (Shrimp)

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Guide to STPM Practicals

Preservation of Plant and Insect Specimens
This experiment enables students to: 1. Learn the skill of preservation of plants and insects prior to related experiments 2. Understand better the field of taxonomy 3. Identify the morphological features of plants and insects 4. Determine the phylum, class and order of the preserved plants and insects Below are some of the plants and insects that can be preserved by dry or wet preservation techniques using FAA solution or 70% ethanol as preservative. Both these techniques can cause the colour of the specimens to fade and therefore their original colour should be noted before preserving them. Insects can be killed by squeezing their thorax or abdomen gently to block their air passages. Both the preservation techniques enable the plants and insects to be preserved for a long time.
Gemma cups Male umbrellas Female umbrellas

Marchantia sp. is a liverwort commonly found in flowerpots in green houses, on moist bricks in gardens and on badly drained soils. On its thalloid body are small gemma cups with small oval pieces of tissue, which can be spread by raindrops and become new plants. This dioecious bryophyte can easily be identified by its male and female umbrellas which carry the male and female receptacles. Marchantia belongs to the division, Bryophyta and class, Hepaticae. Dryopteris sp. which belongs to the division, Filicinophyta and class, Filicinae is a common fern found growing in dry open area. It is a hardy plant which reproduces rapidly and asexually by means of spores found in the sporangia that grow in clusters called sori on the underside of its fronds. The fern is also capable of reproducing sexually.

or

Both the housefly and the dragonfly or grasshopper (or locust) belongs to the phylum, Arthropoda which has the characteristics of segmented body, jointed legs in pairs, chitinous exoskeleton and open circulation. Both the animals belong to the class Insecta, which can be characterized by their bodies being divided into 3 distinct regions, that is, the head, thorax and abdomen. The head bears a pair of antennae, the thorax bears 3 pairs of jointed legs and the body is well supplied with respiratory tubes or trachea. The housefly belongs to the order Diptera, whereas the grasshopper/ locust belongs to the order Orthoptera. Dragonflies, however belong to the order Odonata.
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Guide to STPM Practicals

Collection of Insects
This experiment enables students to: 1. Be aware of the diversity of insects in Malaysia 2. Learn the insects habitats, feeding and sexual behaviour, and their economic values 3. Strengthen their concept and understanding of classification and nomenclature 4. Inculcate their love towards all living things and nature Capturing of insects and preserving them 1. Insects can be captured by: (a) Sweeping small bushes with a net (b) Catching flying insects like butterflies directly with a net (c) Using a light trap to catch nocturnal insects 2. The captured insects are then placed into the killing jar containing cotton wool soaked in concentrated ethanol. After the insects have been fully paralyzed, they are pinned on a piece of polystyrene to fix the position. 3. With suitable dissecting tools, the abdomen of the insect is slit open and the internal organs were removed. A wad of formalin-soaked cotton wool is then inserted into the abdominal cavity (see diagrams below). 4. Formalin is also injected into parts which are too small to be cut. The slit is then sealed.

The abdomen of the insect is slit open and the internal organs are removed

A wad of formalin-soaked cotton wool is inserted into the abdominal cavity

5. The preserved insects are then clearly displayed and pinned onto a polystyrene. Each insect is labeled as follows: Local name: Order: Location: Habitat: Date of collection: Collectors name: (A total of 10 different species from ten different orders of insects are to be collected) 6. A suggested list of insects is given below. (a) Local name: Longhorn beetle Scientific name: Batocera davidis Order: Coleoptera (b) Local name: Brown spruce longhorn beetle Scientific name: Tetropium fuscum Order: Coleoptera
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 (c) Local name: Titanic stag beetle Scientific name: Dorcus titanic Order: Coleoptera (d) Local name: Elephant beetle Scientific name: Xylotrupes ulysses Order: Coleoptera (e) Local name: Click Beetle Scientific name: Limonius canus Order: Coleoptera (f) Local name: Sloe bug Scientific name: Dolycoris baccarum Order: Hemiptera (g) Local name: Carpenter bee Scientific name: Xylocopa violacea Order: Hymenoptera (h) Local name: Carpenter bee Scientific name: Xylocopa aruana Order: Hymenoptera (i) Local name: Honey bee Scientific name: V espa affinis Order: Hymenoptera (j) Local name: Peacock pansy Scientific name: Junonia almana Order: Lepidoptera (k) Local name: Striped albatross Scientific name: Appias libythea olferna Order: Lepidoptera (l) Local name: Paper kite Scientific name: Idea leuconoe Order: Lepidoptera (m) Local name: Great orange-tip butterfly Scientific name: Hebomoia glaucippe Order: Lepidoptera (n) Local name: Pepatung merah Scientific name: Nannophya pygmaea Order: Odonata (o) Local name: Ruby meadowhawk dragonfly Scientific name: Crocothemis servilia Order: Odonata (p) Local name: Red grasshawk dragonfly Scientific name: Neurothemis fluctuans Order: Odonata (q) Local name: Common skimmer Scientific name: Neurothemis intermedia Order: Odonata (r) Local name: Blue dragonfly Scientific name: Trithemis festiva Order: Odonata
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 (s) Local name: Common field grasshopper Scientific name: Chorthippus brunneus Order: Orthoptera (t) Local name: Short horned grasshopper Scientific name: Chorthippus parallelus Order: Orthoptera (u) Local name: Grasshopper Scientific name: V alanga nigricornis Order: Orthoptera (v) Local name: Katydid Scientific name: Pterophylla camellifolia Order: Orthoptera (w) Local name: Sawfly Scientific name: Arge humeralis Order: Hymenoptera (x) Local name: Stick insect Scientific name: Phasma reinwarditi Order: Phasmida (y) Local name: Cicada Scientific name: Cicadetta montana Order: Homoptera 7. Some interesting insects that are found around your neighbourhood.

Local name: The wood nymph Order: Lepidoptera

Local name: Stick insect Order: Phasmida

Local name: Stag beetle Order: Coleoptera

Local name: Red dragonfly Order: Odonata

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Guide to STPM Practicals

Collection of Plants
This experiment enables students to: 1. Be aware of the plant diversity in Malaysia 2. Inculcate the love for plants and nature 3. Strengthen their understanding of classification of plants and nomenclature 4. Learn the economic values of Malaysian plants Collection and preservation process 1. Plant specimen to be collected should not be too young or too old and cut to the size to fit a tabloid newspaper. Only the leaf and stem (flowers if any) should be collected. 2. Specimens are pressed immediately after collection and put into the oven at 105 C to rid them of water (see pictures below).

The selected specimen immediately after collection.

is

pressed

The specimen is then put into the oven at 105 C to dry it.

Dried stems and leaves are then mounted.

3. The dried specimen is then mounted onto drawing papers of size 6 in. 10 in. or 13 in. 8 in. 4. Each plant specimen is fully labelled as follows: Local name: Family: Location: Habitat: Date of collection: Collectors name: (A total of 10 different species from ten different families of plants are to be collected) 5. A suggested list of plants that can be collected: (a) Local name: Asoka Scientific name: Saraca indica Family: Fabaceae (b) Local name: Sial menaun Scientific name: Pternandra coerulescens Family: Melastomataceae (c) Local name: Pandan jepun Scientific name: Pandanus amaryllifolius Family: Pandanaceae (d) Local name: Ubi kayu Scientific name: Manihot utilissima Family: Euphorbiaceae
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(e) Local name: Angsana Scientific name: Pterocarpus indicus Family: Papilionaceae

(f) Local name: Tecoma Scientific name: Tabeayaia pentaphylla Family: Bignoniaceae (g) Local name: Beringin/ Malayan Banyan Scientific name: Ficus microcarpa (van nitida) Family: Moraceae (h) Local name: Pulai Scientific name: Alstonia scholaris Family: Apocynaceae (i) Local name: Kelat paya Scientific name: Eugena papilosa Family: Myrtaceae (j) Local name: Gapis Scientific name: Saraca thaipingensis Family: Caesalpiniaceae (k) Local name: Saga Scientific name: Adenanthera pavonina Family: Fabaceae (l) Local name: Melinjau Scientific name: Gnetum gnemon Family: Gnetaceae (m) Local name: Duku Scientific name: Lansium domesticum Family: Meliaceae (n) Local name: Durian Scientific name: Durio zibethinus Family: Bombacaceae (o) Local name: Gelam Scientific name: Melaleuca leucadendron Family: Myrtaceae (p) Local name: Gaharu Scientific name: Aquilaria malaccensis Family: Thymelaeaceae (q) Local name: Gajus Scientific name: Anacardium occidentale Family: Anacardiaceae (r) Local name: Mango Scientific name: Mangifera indica Family: Anacardiaceae
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(s) Local name: Cempedak Scientific name: Artocarpus champeden Family: Moraceae
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 (t) Local name: Berangan Scientific name: Castanopsis inermis Family: Fagaceae (u) Local name: Penaga lilin Scientific name: Mesua ferrea Family: Clusiaceae (v) Local name: Nangka Scientific name: Artocarpus heterophyllus Family: Moraceae (w) Local name: Ixora Scientific name: Ixora javanica Family: Rubiaceae (x) Local name: Kayu manis hutan Scientific name: Cinnamomum iners Family: Lauraceae (y) Local name: Badam Scientific name: Prunus spp. Family: Rosaceae 6. Some interesting plants that can be collected.

Lidah buaya Aloe vera

Cekur Kaempferia galanga

Hempedu bumi Andrographis paniculata

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Guide to STPM Practicals

Ecological Study of a Terrestrial Ecosystem
This experiment enables students to: 1. Learn the basic principles of ecology through hands-on experience 2. Understand the biotic and abiotic elements of ecosystems 3. Understand the dynamic relationship of elements and energy flow in an ecosystem 4. Learn the simple instrumentations (improvised or otherwise) in ecological studies 5. Learn the methods of collecting and analysing ecological data 6. Write a systematic ecological study report 7. Inculcate the love for nature 8. Inculcate good moral values, independence and self-confidence Some of the useful ecological tools that you may use in the course of your study.

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Determination Of Plant Population Using A Quadrat Sampling Technique

1. Students may carry out this study in groups of 4 or 5 but individual report is required. 2. Each group should determine the area to be studied, the objectives, rough working plan, and the techniques to be carried out. 3. The plant distribution of a terrestrial habitat can be determined by using quadrats. 4. The results can be tabulated (see tables and examples below) and calculations should be done using the formulae given below. Quadrat sampling of plants in an open area

An open area (Notice the laterite soil)

Quadrat size: 0.5 m2 (Random sampling)

Formula: (a) Species frequency = no of quandrat containing the species x 100 total number of quandrats Species x frequency Total frequency of all species 100

(b) Relative species frequency = (c) Species density =

Total number of the species x (Total number of quadrat x area of each quadrat) Species x density Total density of all species 100

(d) Relative species density = (e) Species coverage =

Total area of coverage of species x 100 Total number of quandrat Species x coverage 100 Total coverage of all species

(f) Relative species coverage =

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Table 1: Presence of plant species Quadrat Plant species (x) Acantha sp. Ischaemum sp. Imperata cylindrica Lycopodium sp. Melastoma malabathricum Eleucine sp. Polygala sp. Bryophyllum sp. Table 2: Number of plant species Plant species Quadrat 1 2 3 4 5 6 7 8 9 13 13 26 30 11 14 7 23 55 20 10 25 6 Total Total Species number of density 10 species x (/m )
2

5 3 3

7 3 3

8 3 3 3

9 3 3

Total number Species of quadrat frequency containing (%) 10 species x 4 9 1 3 6 4 5 2 2 33 40 90 10 60 40 50 20 20 330

Relative species frequency (%) 12.12 27.27 3.03 18.18 12.12 15.15 6.67 6.67

3 3 3

3 3 3 3 3 3 3 3 3

3 3

Total

Relative species density (%) 15.23 35.00 1.28 9.88 4.88 29.42 3.14 1.16

56 49 Acantha sp. Ischaemum sp. 15 55 50 5 60 32 28 Imperata cylindrica 8 Lycopodium sp. 30 10 20 3 Melastoma 5 7 malabathricum 70 98 Eleucine sp. 5 Polygala sp. 4 Bryophyllum sp. Table 3: Area of coverage

131 301 11 85 42 253 27 10 860

52.4 120.4 4.4 34.0 16.8 101.2 10.8 4.0 344.0 Total area Species of coverage coverage of species x (%) (cm2) 2100 21000 5720 57200 220 2200 15900 6900 52800 8500 2000 166500

Quadrat (area covered by each spp, cm2) Plant species 1 2 3 4 5 6 7 8 100 520 220 280 70 1500 400 800 120 460 200 9 100 600 10

1000 900 Acantha sp. Ischaemum sp. 150 1000 1000 50 1200 640 560 Imperata cylindrica Lycopodium sp. 600 200 400 30 80 Melastoma 90 70 malabathricum Eleucine sp. 980 2200 Polygala sp. 50 80 Bryophyllum sp.

Relative species coverage (%) 12.61 34.35 1.32 9.55 4.14 31.71 5.11 1.20

1590 690 5280 850 200 16650

Total

The plant species that shows the highest species frequency / species density / species coverage is the most dominant species in the habitat.
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Soil Analysis
1. Soil sampling technique Metal cylinders or tin cans (improvised tools) are pressed or hammered into the soil to collect the soil sample. The soil sample is taken back to the school science laboratory in the cylinder for further analysis (see picture below).

Soil sample in metal cylinder

Laterite soil from open area

Soil colour hints its content

2. Determination of soil texture Students may use a soil sieve or measuring cylinder to determine the percentage of each soil component as shown below.

Sieves of different mesh sizes are used to determine the amount of clay, silt and sand in the soil sample. A measuring cylinder (on the right) can also be used for the same purpose. Calculation of the percentage of components of soil sample Formula Percentage of soil component height of soil component = total height of soil sample Working: Percentage of clay 5.55 = 9.30 100% = 59.67 % Percentage of silt 1.87 = 9.30 100% = 20.11 % Percentage of sand 1.88 = 9.30 100% = 20.22 % Texture of soil clay silt sand Total Height (cm) 5.55 1.87 1.88 9.30 % of soil component 59.67 20.11 20.22 100.00
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100% (using measuring cylinder method)

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CL AY

T EN RC PE

PE RC EN T

LT SI

PERCENT SAND

3. Determine of soil sample pH A sample of soil is added to a test tube contain a spatula of barium sulphate which helps to 3 precipitate the clay particles in a soil sample suspension. The test tube is then filled with 4 water and shaken after adding 5-6 drops of Universal Indicator. A clear coloured liquid is formed above the soil which is noted and the pH recorded as shown in Table 4. The pH of the soil is then correlated to the plant species that is most suitable to live in this pH condition. Table 4: Soil sample 1 2 3 4 5 Colour red red yellow red yellow Average pH pH 4 4 6 5 6 5

The colour of the liquid above the soil sample is compared with the Universal Indicator Chart to determine the pH of the soil.

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Use of the 2 Test in Monohybrid and Dihybrid Genetic Crosses

This experiment enables students to use the 2 test to determine whether a set of experimental data obeys Mendels first and second law. The monohybrid ratio 3:1 and dihybrid ratio 9:3:3:1 are hypothetical estimations based on dominant and recessive genes, segregation, independent assortment and random fertilisation which may be influenced by chance events and will be subjected to normal deviation. To assess a genetic hypothesis, a 2 test was used which can change the deviation from the expected value/ ratio to the probability that chance alone could be responsible for the deviation. This test takes into account the sample size and the number of parameters (degree of freedom). For most problems in genetics, the degree of freedom (df) is one less the number of classes of phenotypes (n). For example, in a monohybrid cross, where only one trait, that is, the colour of maize seed (purple which is dominant over yellow) is considered (n = 2), then the df is n 1 = 2 1 = 1. In a dihybrid cross, where two traits, that is, the colour of maize seed (purple and yellow) and the conditions of maize seed (smooth which is dominant over wrinkled) are considered (n = 4), then the df is n 1 = 4 1 = 3. In this experiment, the phenotypes of the maize seeds are counted from the ears of corn given as below and the results tabulated. 1. Ears of corns from monohybrid crosses (a) The phenotypes: purple and yellow seeds (b) The phenotypes: smooth and wrinkled seeds

Purple seed

Yellow seed

2. Ears of corns from dihybrid cross The phenotypes: purple / yellow and smooth / wrinkled seeds.

Purple wrinkle seed Purple smooth seed Yellow wrinkle seed Yellow smooth seed

From the results, the values of 2 are calculated and compared with the values given in the table below. (o e) 2 = (e)
2

o = observed value, that is, the actual number of seeds of a particular phenotype e = expected value, that is, the number of seeds of a particular phenotype calculated from the monohybrid and dihybrid ratios
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2 table df\p 1 2 3 4 .99 .0201 .115 .297 .98 .0404 .185 .429 .95 .0039 .103 .352 .711 .90 .016 .211 .584 1.064 .80 .064 .446 1.005 1.649 .70 .148 .713 1.424 2.195 .50 .455 1.386 2.366 3.357 .30 1.074 2.408 3.665 4.878 .20 1.642 3.219 4.642 5.989 .10 2.706 4.605 6.251 7.779 .05 3.841 5.991 7.815 9.488 .02 5.412 7.824 9.837 .01 6.635 9.210 11.341 .00016 .00063

11.668 13.277

p Probability df Degree of freedom Monohybrid cross (Mendels first law) Phenotype Expected Observation Expected ratio (o) number of count (e) 1 3 Total 79 212 291
2

Divergence (o e) 6.25 6.25

Divergence2 Divergence2/ (o e)2 Expected no. of count (o e)2/(e) 39.0625 39.0625 0.537 0.179 0.716 = 0.716

Yellow Purple

72.75 218.25

Conclusion: The calculated 2 value (0.716) is found to be lower than the value (3.841) given in the 2 table (P0.05, df = 1). Therefore the deviation is not significant and the result obeys Mendels first law. Dihybrid cross (Mendels second law) Phenotype Expected Observation ratio (o) 9 3 3 1 669 232 231 68 1200 Expected Divergence number of (o e) count (e) 675 225 225 75 6 7 6 7 Divergence2 Divergence2/ (o e)2 Expected no. of count (o e)2/(e) 36 49 36 49 0.053 0.218 0.160 0.653 1.084 2 = 1.084 Conclusion: The calculated 2 value (1.084) is found to be lower than the value (7.815) given in the 2 table (P0.05, df = 3). Therefore the deviation is not significant, the dihybrid ratio of 9:3:3:1 is accepted and the result obeys Mendels second law.

Purple smooth Yellow smooth Purple wrinkled Yellow wrinkled Total

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Guide to STPM Practicals

Extraction of Plant DNA
This experiment enables students to: (a) To indicate the presence of DNA in living cells. (b) To describe the physical characteristics of DNA. Deoxyribonucleic acid (DNA) is a nucleic acid that contains the entire genetic information of an organism is found in its nucleus. DNA is formed from nucleotide subunits namely thymine (T), adenine (A), guanine (G) and cytosine (C). In a cell, DNA exists in the form of double-stranded helix molecule of polynucleotide where there is base-pairing between A and T, and G and C. These two strands run in opposite directions to each other and are anti-parallel. A segment of DNA that carries the genetic information is called a gene. In this experiment, students will use finely-crushed onion succulent leaves and a DNA Extraction Kit (see below) to help them to extract DNA from the onion leaves following the instructions given. The onion leaves must first be homogenized to break down the cell wall as to release the DNA from the plant cell. The isopropanol is used to precipitate the DNA extracted. In a proper forensic investigation, DNA in a solution can be visualized using the electrophoresis method.

At the end of the experiment, a whitish, sticky and slimy DNA is extracted and precipitated at the tip of a pooling stick.

A whitish, sticky and slimy DNA is extracted and preciptated at the end of a pooling stick.
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