C1.

3 Photosynthesis focuses on how light energy is transformed into chemical

energy, which is then used to produce carbohydrates.

1. Light Absorption and Photosystems (C1.3.9, C1.3.10, C1.3.11)

●​ Light energy is absorbed by pigments in photosystems, which are

protein-pigment complexes on the thylakoid membranes.
●​ Photosystem II captures light and uses its energy to split water molecules
(photolysis), releasing:
○​ Oxygen (O₂) as a by-product.
○​ Electrons, which are passed along the electron transport chain.

2. ATP and NADPH Production (C1.3.12, C1.3.13)

●​ The movement of electrons through the electron transport chain pumps

protons into the thylakoid space, creating a proton gradient.
●​ Protons flow back through ATP synthase, producing ATP (chemiosmosis).
●​ Electrons reduce NADP⁺ to NADPH.

3. Carbon Fixation and the Calvin Cycle (C1.3.15 to C1.3.18)

●​ In the Calvin cycle, ATP and NADPH are used to convert CO₂ into glucose.
1.​ Carbon Fixation (by Rubisco): CO₂ is attached to a 5-carbon
compound (RuBP).
2.​ Reduction Phase: ATP and NADPH convert the 3-carbon compound
into G3P (a sugar precursor).
3.​ Regeneration of RuBP: ATP is used to regenerate RuBP, enabling the
cycle to continue.

4. Role of Abiotic Factors (C1.3.8)

●​ Photosynthesis rates are influenced by:

○​ Light intensity (more light = more ATP and NADPH production).
○​ Carbon dioxide concentration (higher CO₂ = more glucose
production).
○​ Temperature (affects enzyme activity in the Calvin cycle).
○​ Water and nutrient availability (affects photosystem efficiency).

5. Experimental Techniques (C1.3.4, C1.3.7, C1.3.8)

●​ Chromatography: Separates and identifies photosynthetic pigments.

●​ Absorption and Action Spectra: Show which wavelengths of light are
absorbed and used for photosynthesis.
●​ Limiting Factor Experiments: Varying CO₂, light, or temperature to observe
their impact on the rate of photosynthesis.
📗 C1.3.1 Transformation of Light Energy to Chemical Energy
●​ What is happening?​
Light energy from the Sun is converted into chemical energy stored in carbon
compounds.
●​ Why is it important?​
These carbon compounds (e.g., carbohydrates, lipids, proteins, nucleic acids)
are essential for life processes such as growth, repair, and energy storage.
●​ How does it happen?
○​ Pigments like chlorophyll absorb light energy.
○​ This energy powers reactions that combine carbon dioxide and water
into sugars.
○​ Chemical energy from these sugars is passed through food chains,
supporting life in ecosystems.

🌿 C1.3.2 Conversion of Carbon Dioxide to Glucose

●​ The Process:​
Plants use hydrogen from water to reduce carbon dioxide into glucose. The
overall reaction is:

●​ Where does hydrogen come from?​

From the photolysis of water (splitting water using light):

●​ 4 electrons (e⁻): Used to drive reactions in the electron transport chain.

●​ 4 protons (H⁺): Combine with CO₂ to form glucose during the Calvin cycle.
●​ Oxygen (O₂): Released as a by-product.

C1.3.3 Oxygen as a By-product of Photosynthesis

Where Does Oxygen Come From?

●​ Oxygen is produced during the photolysis of water in photosystem II.

●​ The reaction is:

 ○​ Electrons (e⁻): Travel through the electron transport chain to produce
ATP and NADPH.
○​ Protons (H⁺): Are used in the Calvin cycle to reduce CO₂ to glucose.
○​ Oxygen (O₂): Is released as a by-product.

🧪 Release of Oxygen
Practical 20.1 Detection of oxygen produced in photosynthesis

●​ The oxygen produced accumulates in the chloroplast.

●​ It diffuses out of the chloroplast, then out of the leaf cells into air spaces
inside the leaf.
●​ Finally, it escapes into the atmosphere through tiny pores called stomata.

In aquatic plants, oxygen is visible as bubbles that rise from the leaves when
water becomes saturated with oxygen.

🦠 First Photosynthesizers
●​ Prokaryotes (cyanobacteria) were the first organisms to perform
photosynthesis, over 3,500 million years ago.
●​ Later, algae and plants evolved photosynthesis using specialized organelles
called chloroplasts, which originated from cyanobacteria through
endosymbiosis.

🧮 Balanced Equation for Photosynthesis

The complete reaction showing oxygen production is:

●​ 12 water molecules are split to produce enough hydrogen ions (H⁺) to form
1 glucose molecule (C₆H₁₂O₆).
●​ 6 oxygen molecules (O₂) are released as a by-product.

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 Photosynthesis (UPDATED)

🌿 What is Photosynthesis?
Photosynthesis is the process where plants, algae, and some bacteria convert light
energy into chemical energy stored in glucose (a type of sugar). This process is
vital for life because it provides both food (glucose) and oxygen, which animals
and humans depend on.

🧪 Overall Equation of Photosynthesis:

●​ Carbon dioxide (CO₂) and water (H₂O) are the reactants (inputs).
●​ Glucose (C₆H₁₂O₆) and oxygen (O₂) are the products (outputs).

🌞 Two Main Stages of Photosynthesis:

1. Light-Dependent Reactions (Happen in Thylakoids):

●​ Location: Thylakoid membranes inside chloroplasts.

●​ Process:
○​ Chlorophyll (a green pigment) absorbs light energy, mainly from red
and blue wavelengths.
○​ Photolysis: Light splits water molecules into:
■​ Electrons (e⁻): Power the production of ATP and NADPH.
■​ Protons (H⁺): Help form glucose later.
■​ Oxygen (O₂): Released as a by-product through stomata (tiny
pores in leaves).
●​ Outputs: ATP and NADPH (energy carriers) and O₂ (waste gas).

2. Light-Independent Reactions (Calvin Cycle – Happen in Stroma):

●​ Location: Stroma (fluid outside thylakoids).

●​ Process:
○​ CO₂ enters through stomata (pores in leaves).
○​ Carbon fixation: An enzyme called RuBisCO helps combine CO₂ with a
molecule called RuBP to start the Calvin cycle.
○​ ATP (energy) and NADPH (electrons) from the light-dependent reactions
power the process.
 ○​ The cycle produces a molecule called G3P, which is converted into
glucose (C₆H₁₂O₆).
●​ Outputs: Glucose, which plants use for energy and building structures.

🧬 Connection to Cellular Respiration:

●​ Photosynthesis and cellular respiration are complementary processes:
○​ Photosynthesis stores energy in glucose.
○​ Cellular respiration breaks down glucose to release energy (ATP).
●​ The outputs of photosynthesis (glucose and oxygen) are the inputs for
cellular respiration, and vice versa.

🌵 Adaptations for Photosynthesis:

Some plants have special adaptations to survive in different environments:

●​ Cacti (CAM Photosynthesis):

○​ Open stomata at night to capture CO₂ and store it.
○​ Perform photosynthesis during the day with stomata closed, saving
water.

Separation and Identification of Photosynthetic Pigments by Thin Layer

Chromatography (TLC)

https://www.youtube.com/watch?v=W56RHxu2Hpc&ab_channel=StephanieCastle

Introduction:​
Chloroplasts contain various pigments, including several types of chlorophyll and
accessory pigments. These pigments absorb different wavelengths of light, which
makes them appear in different colors. Thin Layer Chromatography (TLC) is an
effective technique to separate these pigments from leaf tissue.

Materials Required:

●​ Fresh green leaf

●​ Mortar and pestle
●​ Sand
●​ Propanone (acetone)
●​ Watch glass
●​ Paintbrush
●​ Thin Layer Chromatography (TLC) strip
●​ Cork or bung with a slot
●​ Specimen tube
●​ Ruler (with millimeter markings)
 ●​ Hairdryer
●​ Pencil

Procedure:

1.​ Tear a leaf into small pieces and place them in a mortar.
2.​ Add a small amount of sand to aid in grinding.
3.​ Pour a small volume of propanone into the mortar.
4.​ Grind the leaf tissue using the pestle to extract pigments.
5.​ If the propanone evaporates, add a little more.
6.​ When the propanone turns dark green, allow the solid particles to settle and
pour off the liquid into a watch glass.
7.​ Use a hairdryer to evaporate the propanone and water from the cytoplasm,
leaving a dry pigment smear.
8.​ Add 3–4 drops of propanone to dissolve the dry pigment.
9.​ Use a paintbrush to apply a small, dark spot of pigment onto the TLC strip, 10
mm from the bottom. Allow each layer to dry before adding another drop to
darken the spot.
10.​ Fit the other end of the TLC strip into the slot of a cork or bung.
11.​ Place the cork and strip into a specimen tube, ensuring the strip nearly
reaches the bottom but does not touch it.
12.​ Mark the outside of the tube just below the spot level on the TLC strip.
13.​ Remove the strip and cork from the tube.
14.​ Pour solvent into the tube up to the marked level (use a fume cupboard or
well-ventilated area).
15.​ Lower the TLC strip back into the tube, ensuring the solvent does not touch
the pigment spot.
16.​ Leave the tube undisturbed for about 5 minutes until the solvent nearly
reaches the top of the strip.
17.​ Remove the strip and separate it from the cork.
18.​ Draw two pencil lines: one at the solvent front and one at the initial pigment
spot.
19.​ Circle each separated pigment spot and mark the center with a cross.
20.​ Measure the distances moved by the solvent and each pigment (from the
lower line to the cross).
Calculation of Rf Values:​
The Rf value is calculated using the formula:

Rf = Distance moved by pigment / Distance moved by solvent

Table of Standard Rf Values:

Pigment Colour Rf Value

Carotene Orange 0.98
Chlorophyll a Blue-green 0.59
Chlorophyll b Yellow-green 0.42
Phaeophytin Olive-green 0.33
Xanthophyll 1 Yellow 0.28
Xanthophyll 2 Yellow 0.15

Conclusion:​
This experiment demonstrates how thin layer chromatography separates
photosynthetic pigments based on their solubility and affinity for the TLC medium.
By measuring and comparing Rf values, we can identify the different pigments
present in the leaf sample.

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Absorption of Specific Wavelengths of Light by Photosynthetic Pigments

The first stage of photosynthesis involves the absorption of sunlight by pigments.

Pigments absorb certain wavelengths of light, and their color corresponds to the
wavelengths they reflect or transmit.

Key Concepts:

●​ Light and Pigments: White substances reflect all wavelengths, and

transparent substances transmit all wavelengths. Black pigments absorb all
wavelengths, converting the energy primarily to heat.
●​ Color Perception: The color we see is the reflected light. For example, a
gentian flower absorbs all wavelengths except blue, which it reflects.

Photon Energy and Pigment Absorption:

●​ Light travels as particles called photons, each carrying a discrete amount of

energy.
●​ Photon energy is inversely proportional to wavelength: shorter wavelengths
have more energy.
 ●​ Pigment molecules absorb photons if the photon energy excites an electron to
a higher energy state.

Photosynthetic Pigments:

●​ Chlorophylls are the main photosynthetic pigments, appearing green because

they absorb photons from the red and blue parts of the spectrum while
reflecting green wavelengths.
●​ Accessory pigments broaden the range of light absorbed, contributing to
photosynthesis.

Absorption Spectrum:

●​ X-axis: Shows light wavelength (400 to 700 nm, the visible range) with colors
indicated.
●​ Y-axis: Displays the percentage of light absorbed.

Understanding how pigments absorb specific wavelengths is essential for

interpreting photosynthesis and plant growth efficiency.