Botany · Photosynthesis in Higher Plants

Photosynthetic Pigments

NCERT Class 11 Biology, Section 11.4 identifies four pigments in the leaves of higher plants — chlorophyll a, chlorophyll b, xanthophylls, and carotenoids — separated and characterized by paper chromatography. This subtopic carries consistent NEET weight: questions test absorption peaks, Rf ordering, the antenna-versus-reaction-centre distinction, and the critical rule that chlorophyll a alone directly drives the light reaction.

NCERT Grounding

NCERT Class 11 Biology, Chapter 11, Section 11.4 ("How many types of pigments are involved in photosynthesis?") establishes the canonical four-pigment model through paper chromatography of leaf extracts. The text states explicitly: "A chromatographic separation of the leaf pigments shows that the colour we see in leaves is not due to a single pigment but due to four pigments: Chlorophyll a (bright or blue green), chlorophyll b (yellow green), xanthophylls (yellow) and carotenoids (yellow to yellow-orange)."

"Though chlorophyll is the major pigment responsible for trapping light, other thylakoid pigments like chlorophyll b, xanthophylls and carotenoids… also absorb light and transfer the energy to chlorophyll a."

NCERT Biology Class 11, Section 11.4

Section 11.5 further specifies that each photosystem has all the pigments (except one molecule of chlorophyll a) forming the light harvesting or antenna system, while the single chlorophyll a molecule forms the reaction centre — the photochemically active core that initiates electron transport.

The Four Pigments — Structure and Function

All four pigments reside in the thylakoid membranes of the chloroplast. They are organized into two functional pools within each photosystem: the large antenna (light harvesting) complex and the single reaction-centre chlorophyll a. The four pigments differ in molecular structure, colour, absorption wavelengths, and role in photosynthesis.

Organizing principle: Chlorophyll a is the only pigment capable of directly donating an excited electron to the primary electron acceptor. Every other pigment transfers absorbed energy to chlorophyll a by resonance (Förster) transfer and is therefore called an accessory pigment.

Chlorophyll a

430 & 680 nm

Absorption peaks (blue-violet + red)

Colour: Blue-green (bright green in chromatogram)

Role: Primary photosynthetic pigment; forms reaction centre (P700 in PS I, P680 in PS II)

Key fact: The most abundant plant pigment on Earth; only pigment that directly participates in the light reaction

NEET Trap — reaction centre only

Chlorophyll b

450 & 640 nm

Absorption peaks (blue + orange-red)

Colour: Yellow-green

Role: Accessory pigment; broadens the spectral range of light harvesting; transfers energy to chlorophyll a

Key fact: Differs from chl a by –CHO group instead of –CH₃ at C-7; more polar than chl a

Accessory — not a reaction centre

Xanthophylls

~450 nm

Absorption peak (blue region)

Colour: Yellow

Role: Accessory pigment; photoprotective — quenches excess energy and scavenges singlet oxygen; energy transfer to chl a

Key fact: Oxygenated carotenoids (contain –OH groups); less non-polar than carotene → lower Rf

Carotenoid class — oxygenated

Carotenoids (β-carotene)

450–480 nm

Absorption peaks (blue region)

Colour: Yellow to yellow-orange

Role: Accessory + photoprotection; quenches triplet chlorophyll and singlet O₂ preventing photo-oxidation of chl a

Key fact: β-carotene is a precursor of Vitamin A; gives carrot its characteristic colour; most non-polar pigment → highest Rf

Highest Rf in chromatography

Absorption Peaks — Comprehensive Table

Pigment Chromatogram Colour Peak 1 Peak 2 Type Rf Order
Chlorophyll a Blue-green (bright) ~430 nm (blue-violet) ~680 nm (red) Primary + reaction centre 3rd (second-lowest)
Chlorophyll b Yellow-green ~450 nm (blue) ~640 nm (orange-red) Accessory 4th (lowest — most polar)
Xanthophylls Yellow ~450 nm (blue) Accessory + photoprotective 2nd
Carotenoids (β-carotene) Yellow-orange 450–480 nm (blue) Accessory + photoprotective 1st (highest — most non-polar)

Antenna Complex vs Reaction Centre

Each photosystem is architecturally divided into two components: the light harvesting complex (LHC), also called the antenna complex, and the reaction centre. This distinction is among the most frequently tested concepts from this section in NEET.

Antenna Complex vs Reaction Centre — key distinctions

Antenna Complex (LHC)

~300

pigment molecules per photosystem

  • Contains all four pigments: chl a, chl b, xanthophylls, carotenoids
  • Functions by absorbing photons across a broad spectrum
  • Transfers excitation energy to the reaction centre by resonance (Förster) transfer — no electrons are moved here
  • Acts as a funnel: hundreds of molecules collect energy and pass it to one special chl a
  • Increases photosynthetic efficiency in low-light environments
VS

Reaction Centre

1

special chlorophyll a molecule

  • Contains only one special chlorophyll a molecule (P700 in PS I; P680 in PS II)
  • Undergoes actual photochemical reaction: excited electron is ejected to the primary electron acceptor
  • In PS II: P680 absorbs red light at 680 nm; electron excitation initiates water splitting
  • In PS I: P700 absorbs red light at 700 nm; excited electron ultimately reduces NADP⁺ to NADPH
  • The photochemical step that converts light energy into chemical energy occurs here

NCERT (Section 11.5) states: "Each photosystem has all the pigments (except one molecule of chlorophyll a) forming a light harvesting system also called antennae. The single chlorophyll a molecule forms the reaction centre." The reaction centre chlorophyll a in PS I is called P700 because it has an absorption maximum at 700 nm; in PS II it is called P680 because its absorption maximum is at 680 nm.

Figure 1 — Antenna and Reaction Centre in a Photosystem Photosystem Antenna Complex and Reaction Centre THYLAKOID MEMBRANE Chl b Carot. Xanth. Chl b Chl a Carot. P700 (or P680) Primary Acceptor e⁻ ejected Photon ANTENNA COMPLEX (LHC) REACTION CENTRE

Figure 1. Schematic of a photosystem. Hundreds of antenna pigment molecules (chlorophyll a, b, xanthophylls, carotenoids — shown as coloured circles) absorb photons and transfer excitation energy by resonance to the single reaction centre chlorophyll a (P700 in PS I; P680 in PS II — black circle). The reaction centre then ejects a high-energy electron to the primary electron acceptor, the only step that constitutes direct participation in the light reaction.

Paper Chromatography and Rf Values

Paper chromatography is the experimental technique used to separate the four leaf pigments. A concentrated leaf extract is spotted on chromatography paper and developed with a non-polar solvent (e.g., petroleum ether : acetone mixture). Pigments migrate at rates determined by their polarity relative to the stationary phase (paper, polar) and the mobile phase (solvent, non-polar).

The Rf (retardation factor) is defined as the ratio of the distance travelled by the pigment to the distance travelled by the solvent front. A more non-polar pigment interacts less with the polar paper and therefore travels farther, yielding a higher Rf value.

Chromatography band order (top to bottom — highest Rf to lowest)

Petroleum ether–acetone solvent system
  1. Band 1

    Carotene (β-carotene)

    Yellow-orange. Highest Rf. Most non-polar pigment — no oxygen-containing groups. Travels farthest.

    Rf ≈ 0.98
  2. Band 2

    Xanthophylls

    Yellow. Second highest Rf. Oxygenated carotenoids (–OH groups) make them slightly more polar than carotene.

    Rf ≈ 0.71
  3. Band 3

    Chlorophyll a

    Blue-green. Third band. Less polar than carotene and xanthophylls but less polar than chlorophyll b.

    Rf ≈ 0.65
  4. Band 4

    Chlorophyll b

    Yellow-green. Lowest Rf. Most polar pigment due to additional –CHO group. Travels shortest distance.

    Rf ≈ 0.45
4

Distinct pigment bands

A standard leaf chromatogram resolves into exactly four bands. NEET questions sometimes ask how many bands appear — the answer is always four, not two (as students sometimes confuse with the two pigment classes: chlorophylls and carotenoids).

· 1

Reaction centre pigment

Only one pigment — chlorophyll a — directly participates in the photochemical step. All others are accessory pigments, regardless of how efficiently they absorb light.

Why accessory pigments matter

The absorption spectrum of chlorophyll a shows strong peaks only in the blue (~430 nm) and red (~680 nm) regions. The green region (around 500–600 nm) is reflected — which is why leaves appear green. Without accessory pigments, this large portion of the solar spectrum would be wasted.

Chlorophyll b absorbs at 450 nm and 640 nm, filling part of the gap. Xanthophylls and carotenoids absorb in the 450–480 nm blue range. Together, the four pigments cover a much wider portion of the visible spectrum, improving the efficiency of light capture in natural conditions. This is the fundamental reason plants possess accessory pigments.

The second function of accessory pigments — particularly carotenoids — is photoprotection. Under high-intensity illumination, chlorophyll a can form reactive triplet states and generate singlet oxygen, which is damaging to the photosynthetic machinery. Carotenoids quench these reactive species, preventing photo-oxidation (photobleaching) of chlorophyll a. NCERT Section 11.4 states this explicitly: accessory pigments "not only enable a wider range of wavelength of incoming light to be utilised for photosynthesis but also protect chlorophyll a from photo-oxidation."

Worked Examples

Worked example 1 — Chromatography band identification

In a paper chromatography experiment on a leaf extract, four bands are observed. Band A is at the top of the chromatogram and Band D is at the bottom. Which of the following correctly identifies the bands?
(1) A = Chlorophyll a, B = Chlorophyll b, C = Xanthophylls, D = Carotene
(2) A = Carotene, B = Xanthophylls, C = Chlorophyll a, D = Chlorophyll b
(3) A = Xanthophylls, B = Carotene, C = Chlorophyll b, D = Chlorophyll a
(4) A = Carotene, B = Chlorophyll a, C = Xanthophylls, D = Chlorophyll b

Answer: (2). The band at the top of the chromatogram has the highest Rf value, meaning it travelled the farthest. Carotene is the most non-polar pigment and has the highest Rf. The order from top to bottom (highest to lowest Rf) is: Carotene → Xanthophylls → Chlorophyll a → Chlorophyll b. Chlorophyll b is the most polar pigment and stays closest to the origin. Options (1), (3), and (4) incorrectly place the pigments.

Worked example 2 — Reaction centre identification

Suppose a plant lacked chlorophyll a but had high concentrations of chlorophyll b and carotenoids. Would it be capable of carrying out photosynthesis? Justify your answer with reference to the antenna complex and reaction centre.

Answer: No. Chlorophyll b and carotenoids are accessory pigments. They absorb light and transfer the energy by resonance to the reaction centre chlorophyll a, but they cannot themselves act as the reaction centre. The photochemical step — ejection of a high-energy electron to the primary electron acceptor — can only be performed by the special chlorophyll a molecule (P700 or P680) at the reaction centre. Without chlorophyll a, no electron transport chain can be initiated, ATP and NADPH cannot be produced, and carbon fixation cannot occur. The plant would be unable to carry out photosynthesis despite having functional antenna pigments. This is an NCERT Exercise Question 5 (Section 11.5).

Worked example 3 — Absorption peak matching

Match the pigment with its correct absorption peak and chromatogram colour:
P. Chlorophyll a — (i) 450 nm + 640 nm, yellow-green
Q. Chlorophyll b — (ii) 430 nm + 680 nm, blue-green
R. Carotene — (iii) 450–480 nm, yellow-orange
S. Xanthophyll — (iv) ~450 nm, yellow
Select the correct match.

Answer: P–(ii), Q–(i), R–(iii), S–(iv). Chlorophyll a absorbs at 430 nm and 680 nm and appears blue-green. Chlorophyll b absorbs at 450 nm and 640 nm and appears yellow-green (it has a longer-wavelength second peak than chlorophyll a's red peak). Carotene absorbs in the 450–480 nm range and is yellow-orange — it gives carrot its colour. Xanthophylls are oxygenated carotenoids absorbing near 450 nm and appear yellow.

Common Confusion & NEET Traps

Chlorophyll a vs Chlorophyll b — the most-confused pair

Chlorophyll a

680 nm

Second absorption peak (red)

  • Blue-green colour; bright green in chromatogram
  • Absorption peaks at ~430 nm and ~680 nm
  • Forms the reaction centre: P680 (PS II) and P700 (PS I)
  • Primary photosynthetic pigment — directly participates in light reaction
  • Less polar → higher Rf (third from bottom)
  • Has –CH₃ group at C-7 of the porphyrin ring
VS

Chlorophyll b

640 nm

Second absorption peak (orange-red)

  • Yellow-green colour
  • Absorption peaks at ~450 nm and ~640 nm
  • Does NOT form the reaction centre — purely accessory
  • Transfers absorbed energy to chlorophyll a; never ejects an electron directly
  • More polar → lowest Rf (travels least in chromatogram)
  • Has –CHO group at C-7 (replacing –CH₃ of chl a) — increases polarity

NEET PYQ Snapshot — Photosynthetic Pigments

Real questions from NEET 2018 — directly relevant to this subtopic. Study the solution logic, not just the answer.

NEET 2018 · Q.101

Which of the following is not a product of the light reaction of photosynthesis?

  1. ATP
  2. NADH
  3. NADPH
  4. Oxygen
Answer: (2) NADH

Why: The light reaction produces ATP (via photophosphorylation), NADPH (by reduction of NADP⁺ at PS I using electrons from water), and O₂ (by photolysis of water at PS II). NADH is produced in cellular respiration (glycolysis, Krebs cycle) — it is not a product of photosynthetic light reactions. The reductant in photosynthesis is specifically NADPH (nicotinamide adenine dinucleotide phosphate, reduced), not NADH. This is a classic NEET confusion: both are electron carriers, but their metabolic contexts are completely different. Students who confuse the two fail this question consistently.

NEET 2018 · Q.130

Oxygen is not produced during photosynthesis by which of the following?

  1. Green sulphur bacteria
  2. Chara
  3. Cycas
  4. Porphyra
Answer: (1) Green sulphur bacteria

Why: Green sulphur bacteria are anoxygenic photosynthesizers — they use H₂S (not water) as their electron donor. Because there is no photolysis of water, no O₂ is released. The oxidation product is sulphur, not O₂. This connects directly to van Niel's generalisation (NCERT Section 11.2). Chara is a green alga (oxygenic), Cycas is a gymnosperm (oxygenic), and Porphyra is a red alga (oxygenic). All three evolve O₂ through water splitting at PS II. Note: all pigment-containing, oxygenic photosynthesizers have chlorophyll a at their PS II reaction centre (P680), which is why the pigments studied in this subtopic are fundamental to understanding which organisms can split water.

Concept

In a paper chromatography experiment separating leaf pigments, four bands appear. Which band is at the top (highest Rf), and which is at the bottom (lowest Rf)?

  1. Top: Chlorophyll a; Bottom: Chlorophyll b
  2. Top: Xanthophylls; Bottom: Carotene
  3. Top: Carotene; Bottom: Chlorophyll b
  4. Top: Chlorophyll b; Bottom: Carotene
Answer: (3) Top: Carotene; Bottom: Chlorophyll b

Why: Carotene is the most non-polar pigment and interacts least with the polar paper stationary phase, so it migrates farthest with the non-polar solvent front — highest Rf, topmost band. Chlorophyll b has the –CHO group making it the most polar of the four pigments; it adheres most to the paper and migrates least — lowest Rf, bottommost band. The complete order is: Carotene > Xanthophylls > Chlorophyll a > Chlorophyll b.

FAQs — Photosynthetic Pigments

Frequently tested conceptual questions from this subtopic — expand each for the full answer.

Which photosynthetic pigment directly participates in the light reaction?

Chlorophyll a is the only pigment that directly participates in the light reaction. It forms the reaction centre in both PS I (P700) and PS II (P680). All other pigments — chlorophyll b, xanthophylls, and carotenoids — are accessory pigments that transfer absorbed energy to chlorophyll a.

What are the absorption peaks of chlorophyll a?

Chlorophyll a shows two major absorption peaks: approximately 430 nm (blue-violet region) and 680 nm (red region). The reaction centre forms in PS II absorb maximally at 680 nm (P680) and in PS I at 700 nm (P700).

What is the order of Rf values in paper chromatography of leaf pigments?

In paper chromatography, carotene has the highest Rf value (travels farthest) because it is the most non-polar pigment. The order from highest to lowest Rf is: carotene > xanthophyll > chlorophyll a > chlorophyll b. Chlorophyll b is most polar and therefore travels the shortest distance.

Why are accessory pigments essential despite not being the reaction centre?

Accessory pigments — chlorophyll b, xanthophylls, and carotenoids — absorb wavelengths of light not efficiently captured by chlorophyll a. They broaden the range of usable wavelengths and funnel the absorbed energy to the chlorophyll a reaction centre. Carotenoids also protect chlorophyll a from photo-oxidation (photobleaching) under intense light.

What is the difference between the antenna complex and the reaction centre?

The antenna complex (light harvesting complex, LHC) consists of hundreds of pigment molecules — all four pigments — that absorb light and channel energy by resonance transfer to the reaction centre. The reaction centre is a single special chlorophyll a molecule that actually undergoes photochemical excitation and donates an electron to the primary electron acceptor, initiating electron transport.

Why does a leaf kept in the dark turn yellow?

Chlorophylls (a and b) are less stable and break down faster than carotenoids when a leaf is kept in the dark and deprived of light-driven regeneration. The yellow-orange carotenoids and yellow xanthophylls persist longer, unmasking their colour as the green chlorophylls degrade.

Can a plant with high chlorophyll b but no chlorophyll a carry out photosynthesis?

No. Chlorophyll b cannot directly donate an electron to the primary electron acceptor because it does not form the reaction centre. It can absorb light and transfer energy, but without chlorophyll a to act as the reaction centre, the photochemical step of light reaction cannot proceed. Therefore, the plant cannot carry out photosynthesis.