Photosynthesis

• ____________ is defined as a complex chemical

process that converts radiant energy (light) to chemical
energy (sugar).
Photosynthesis
• _______ is released to the environment, and sugar is
used to fuel growth or is stored as starch, a
polysaccharide.

• Virtually all organisms are dependent upon

photosynthesis for food and oxygen.

Photosynthesis Photosynthesis
• Photosynthesis occurs in the following 3 stages: 1.) capturing
• A Summary equation for photosynthesis is demonstrated energy from sunlight 2.) using the energy to make ATP nad
below: reducing power in the form of NADPH 3.) using ATP &
• Equation: NADPH to power synthesis of organic molecules from CO2 in
• light
the air.
• 6 CO2 + 12 H2O C6H1206 + 6 H20 + 6 02
chorophyll Sugar • A.) The first two stages can only take place in the presence of
• carbon dioxide water water oxygen

light which are referred to as the photochemical rxn’s or the

• Therefore, another way to define photosynthesis is the
light-dependent and chlorophyll-dependent conversion of
carbon dioxide and water to sugar, water, and oxygen.
light-dependent rxn’s.
• B.) The 3rd stage occurs via the Calvin Cycle. As long as ATP &
NADPH are available, The Calvin Cycle can occur with or
without light; therefore, these rxn’s are referred to as
Biochemical Rxn’s or the light-independent rxn’s.

Photosynthesis
• light
• 6 CO2 + 12 H2O C6H1206 + 6 H20 + 6 02
chorophyll Sugar water
• carbon dioxide water oxygen

• ** The water molecules seen on both sides of the

equation ARE NOT THE SAME!!!

• ~ _______ H2O molecules split and release electrons

during photochemical rxn’s.
• ~ _______ H2O molecules are formed from Hydrogen
& O2 release during photochemical & biochemical rxn’s.

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 Photosynthesis Photosynthesis
Photochemical Biochemical • All organisms (including plants) carry out
“Light” Rxn’s “Dark” Rxn’s respiration in one form or another.
• Fast (practically • Slower, but still
instantaneous) extremely fast
• Light – dependent • Light - independent • ________-containing organisms (plants) can
• Splits water to release • Converts (fixes) carbon also carry out photosynthesis.
Oxygen, electrons, & dioxide to sugar
protons
• We will start our study of photosynthesis by
explaining the pigments observed in
photosynthesis.

Photosynthesis Uptake of CO2 during photosynthesis

• Pigment: ___________________ • Carbon dioxide will _______ the pH of a plant during

the light-independent reactions of photosynthesis.
• Primary pigments for photosynthesis: (acid)
– chlorophyll a ( blue-green) • As the plant fixes or uses the carbon dioxide, the pH will
– chlorophyll b (yellow-green)
_______ back up toward neutral. (base)
Equation:
• Accessory pigments: H2O + CO2 H2CO3 H+ + HCO3-
– ________ (yellow) – Change to this color on your sheet! Water Carbon Carbonic Hydrogen Bicarbon-
– ________ (yellow-orange) – Change to this color too! Dioxide Acid Ion ate Ion
– * The accessory pigments absorb light and transfer energy to
chlorophyll a.

Use of Light and Chlorophyll to

Produce Starch During Photosynthesis Chloroplast
- The light-dependent reactions of photosynthesis
occur on photosynthetic membranes.

- In plants and algae, these membranes are called

_________, which are located within a special
organelle called a ________ (see Fig. 12.8).

- Thylakoids are stacked to form columns called

_____, which are held in place by ______.
• 1 = cell wall, 3 = vacuole, 4 = chloroplast envelope (2
- A semiliquid called _____ bathes the interior of membranes), 5 = tonoplast, 7 = grana, 8 = stroma
the chloroplast and contains the enzymes that thylakoids, 9 = starch grains, 2 = cytoplasm, 6 = plasma
catalyze the light-independent reactions. membrane and 10 = stroma

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Use of Light and Chlorophyll to Produce
Starch During Photosynthesis
- Sugars produced by photosynthesis are often
stored as ______.
- To produce starch, photosynthesis requires light as
an energy source. In the absence of light, sugars
and starch are not produced.
- Photosynthesis also requires chlorophyll to
capture light energy. Without chlorophyll, sugars
and starches will not be produced.
- You will be able to detect the presence of starch by
staining it with a solution of iodine.
Paper Chromatography
• Paper chromatography: a technique for
separating dissolved compounds such as
chlorophyll, carotene, and xanthophyll
Paper Chromatography
• Separation of
Photosynthetic
pigments using
Paper
Chromatography
Lab Experiments
Paper Chromatography
• Technique: A solution of pigments is applied to the
end of a strip of chromatography paper and the end
is dipped into a solvent. The solvent is absorbed and
as it moves up the paper, the pigments dissolve in
the moving solvent. Some pigments can keep up
with the solvent, some are slower. This differential
movement is due to each pigment’s solubility and
characteristic tendency to stick (adsorb) to the
fibers of the paper. A pigment’s molecular size,
polarity, and solubility determine the tendency to
stick.
• *Pigments that adsorb strongly move the slowest.
Paper Chromatography
• Each pigment will move a different distance up
the chromatography paper due to its own
solubility, adsorbance, polarity, size, and
molecular density
• Absorption spectrum- range of absorption for
each pigment
• Action spectrum- range of light used by plant;
combination of all pigments together so it is
bigger
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 Rf Value
• The relationship of the distance moved by
a pigment to the distance moved by the
solvent front is specific for a given set of
conditions. We call this the Rf number.
• The Rf number for a pigment is constant in
a particular solvent-matrix system
Procedure 12.1
• You will work in PAIRS.
• Each pair will need: collard green, a quarter, a piece of
chromatography paper, a test tube, and a cork stopper.)
• Do not put your fingers all over the paper! Hold it by
the edges so you do not get the oils from your
hands on the face of it.
Procedure 12.1
• Place the chromatography paper (ANGLED TIP
DOWN) in the test tube. Make sure it is straight and
is touching the solvent.
• With a few centimeters of paper sticking out of the
tube, wedge the cork stopper into the tube. This will
hold the paper upright in the tube. Place the tube in
a test tube rack.
• After the solvent front reaches the top of the paper
(about 20 minutes), we will remove our strips and
allow them to dry. Be sure to put a pencil mark on
your solvent front at the top. It will disappear when
it dries.
Calculating the Rf value
• Rf = Distance moved by a pigment
• Distance from pigment origin to solvent front
• We will use paper chromatography to identify each
pigment by its characteristic Rf.
• The four pigments we should see are:
• 1.) Carotenes – an orange band
• 2.)Xanthophylls - a yellow-orange band
• 3.) Chlorophyll a - a blue-green band
• 4.) Chlorophyll b - a yellow-green band
Procedure 12.1
• Use a pencil to mark a line across the paper
about 2 cm up from the ANGLED TIP of the
paper.
• Place the spinach leaf across the paper and roll
the quarter edge across the leaf in line with your
mark on the paper. You may want to reposition
the spinach leaf several times and roll across it in
order to get enough pigment on the paper.
Procedure 12.1
• ~ Mark the distance the
solvent moved
• ~ Mark the distance
each pigment moved
• ~Calculate Rf for each
pigment (distance the
pigment moved/
distance the solvent
moved)
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 Procedure 12.1
• After we observe the dried paper and the
pigments:
• 1.) Label Fig. 12.5 (chromatogram)
• 2.) Calculate the Rf for each pigment and record
it in Table 12.1
Procedure 12.4
• We will follow the directions in the lab book on
p.131 exactly.
• We will allow the Elodea to be exposed to light
for 30-60 minutes.
• After the light exposure, we will use pH papers to
detect differences in the control and Elodea,
especially if color differences are not obvious.
Absorption Spectrum of Chlorophyll
• After “blanking” the spectrophotometer, set the
wavelength at 350nm in absorbance mode.
• The filter lever (a small lever on the front left of the
machine) should be set in the lower range.
• This lever will need to be moved over to the higher range
when you reach wavelength measurements of 600nm.
• Use the 8 wavelength readings in Table 7.5 on p.74 and
read the absorbance of chlorophyll from the spinach for
each wavelength.
• Don’t forget to “blank” the spectrophotometer before each
new wavelength reading!!
• * Refer to Procedure 7.4 on pg. 72
Procedure 12.4
• Phenol red is a pH-indicator that turns yellow in the
presence of an acidic solution (pH < 7) and red in a
neutral or basic solution (pH > 7).
• In this experiment we will use phenol red to detect the
uptake of CO2 by a photosynthesizing aquatic plant,
Elodea.
• We will put the Elodea in a slightly acidic solution made
by your breath and phenol red.
Absorption Spectrum of Chlorophyll
• Collard greens have been ground and filtered in 80%
acetone. The filtrate (our filtered solution of spinach and
acetone) is a light green color.
• We will use this filtrate to determine the absorption
spectrum of chlorophyll using the spectrophotometer.
• The spectrophotometers will be “blanked” with a cuvette
containing 80% acetone.
Procedure 12.6…p.132; Procedure
12.7…p.134
Procedure 12.8…p.134
• We will do this procedure and the rest as 3
demos done by trustworthy and willing
students!
• We will follow the directions exactly for
each procedure.
• When we are finished, record your results
(sketches) in Fig. 12.9a on p.133.
• Answer questions for all procedures on
p.134.