4.

Photosynthesis

4.1. Light and Photosynthesis

Q. WHAT IS PHOTOSYNTHESIS?
1
 Light
 is like a rain of photons of d/t frequencies

 eyes are sensitive to visible light region of the Electromagnetic Spectrum.

 light has the properties of both waves and particles

 White light is a mixture of d/t colors with different wavelengths (VIBGYOR)

Electromagnetic Spectrum and Visible Light 2

How Light is Absorbed

3
Why are plants look GREEN?

4
Plastids
Plastids are cytoplasmic organells of plant cell

Plastids are three types

Leucoplasts

Chromoplasts &

Chloroplasts

5
Leucoplasts
 are colorless/non-pigmented plastids

 usually present in cells that donot recieve direct light

 Are used to store substances.

Examples.

 Amayloplast  Starch

 elaioplasts  Oil /lipids

 Proteinoplasts or Aleuroplasts  Protiens

6
Chromoplasts
• are pigmented plastids
• are non chlorophyllous
• Are used to manufacture and store carotenoids
• Red, orange, brown/ yellow
e.g., carotenes,
xanthophyllus,
fucoxanthin,
phycoerythrin
So, they are used to give color for fruits, flower, roots and
leaves
7
Chloroplasts
 Chloroplast is a site of photosynthesis
 trap light energy & convert it to chemical energy
 Common and most vital plastids
 distribute equally all over the cytoplasm
 Chloroplast contains its own DNA, RNA & ribosomes
 Many of the chloroplast proteins are products of chloroplast
 Others are encoded by nuclear DNA, & imported into the
chloroplast

 chlorophyll is contained within chloroplasts

 Chloroplast has two structures: Grana and Stroma 8

Chloroplasts

9
10
 Photosynthesis
 In Photosynthesis,
 light energy is captured by chlorophyl
 light energy → mechanical energy chemical energy

 ―Biosynthesis of C-cpd from CO2 & H2O by illuminated green cells, H2O & O2 as by-product
 Photosynthesis is a REDOX rxn i.e.,
 photochemical oxidation (H2 donor)
 reduction mechanism (CO2)

11
 Cont’d
Photosynthesis

12
• Photosynthesis has two parts/stages
1-Light Rxn ( photo part)
2- Dark Rxn ( synthesis part)

13
 1- Light dependent phase/ light rxn
 Occurs in grana/ thylakoid membrane.
 Also called ‘Photochemical’ phase, B/c it includes
 light absorption by chlorophyll
 photolysis of water/ splitting water in to es +H+ +O2

 produces oxygen, ATP & NADPH

are end products
 pigments are organized into two discrete photochemical Light
Harvesting Complexes (LHC)
 Photosystem I (PS I)

 Photosystem II (PS II)

*A reaction centre in PS I has absorption peak at 700 nm, hence it is called P700

*PS II has absorption maxima at 680 nm, and called P680

*The synthesis of ATP in photosynthesis is named as Photophosphorylation.

  The synthesis of ATP is named as Photophosphorylation

 non-cyclic photophosphorylation

Z scheme of light reaction

15
Cyclic phosphorylation

 When only PS I is functional, e- is circulated within PS I &

phosphorylation occurs due to cyclic flow of electrons
16
Cyclic
photophosphorylation
Non cyclic
photophosphorylation
only PSI is involved both PSI & PSII are involved

PSI gets e- in cyclic fashion PSI gets e- from PSII &

PSII from water

only ATP molecules are both ATP & NADPH molecules

formed are produced

No H2O, hence no O2 O2 evolved, H2O participates

17
2-Light independent phase
 Occurs in the stroma
 Also known Dark Rxn = Calvin cycle

 C-fixation stage/synthesis stage

 used for CO2 fixation and synthesis
of glucose

18
Calvin cycle proceeds in three stages

i. Carboxylation

ii. Reduction

iii. Regeneration

19
i. Carboxylation

 CO2 fixation into a stable organic intermediates by

1O CO2 Acceptor (RUBP) .

i.e., 6 RuBP + 6 CO2 →2(3-PGA) molecules

 The process is catalysed by RuBisCO or PEPcase

to form 2molecule of 3-PGA

20
ii. Reduction
 a series of reactions that lead to the formation of
glucose

 steps involve the utilization of

 2 molecules of ATP for phosphorylation and

 2 molecules of NADPH for reduction per CO2

molecule fixed
PGA → Glyceraldehyde 3-Phosphate (G3P)

21
Overall reaction of Reduction

22
iii. Regeneration
 Regeneration of RuBP/ PEP is crucial for the cycle
to be continued in uninterrupted manner.

 The regeneration steps require one ATP for

phosphorylation to form RuBP/ PEP
10 of 12 G3P →6 RuBP molecules
2 of 12 G3P →1 Glucose molecule

23
24
 Three ways of C-fixation

 C3-cycle

 C4-cycle

 CAM cycle

25
C3 Pathways/C3 cycle
 The 1O CO2 acceptor is Ribuluse 1,5-Bisphosphate
 Use an Enzyme Called RuBisCO to fix CO2
 RubisCO is the most abundant enzyme
 This occurs in the mesophyll cells
 Palisade or Spongy mesophyll cell
 The 1st cpd formed is Phosphogylceric Acid (PGA-a 3C-cpd)

 Called C3 Plants because … 3C product (PGA)

 PGA ready for sugar formation

E.g., Wheat, Cotton, Tobacco, Legumes, Tomatoes, Potatoes 26

 Cont’d
Photosynthesis: Dark Rxn

What is Calvin cycle?

27
 The main stages of the light-independent reactions are:
• carbon dioxide reacts with ribulose bisphosphate (RuBP)

• the reaction is catalysed by the enzyme Rubisco.

• two molecules of the three-carbon compound GP are formed

• each molecule of GP is converted to TP (triose phosphate –3C)

 this is a reduction reaction using hydrogen ions from reduced NADP and
energy from ATP

• some of the TP formed is used to regenerate the RuBP (ATP is

again required)

whilst some is used to form glucose and other useful organic compounds.

28
 summary of the light-independent reactions
 three ‗turns of the cycle‘ result in an
output of one molecule of GP/TP.
 Six turns of the cycle would give an output of two
molecules of TP – enough to make one molecule of
glucose.
 GP/TP can also be converted to lipids, amino acids and
from these into nucleotides and all the other organic
molecules found in plants.
 TP is the basis for the synthesis of all organic molecules.

29
C4 Pathways /‘Hatch-Slack Pathway’

 The firstCO2 acceptor is Phosphoenolpyruvate (PEP).

 Use PEPcase for CO2 Fixation in Mesophyll Cells

 PEPcase has a much higher affinity for CO2 than does RubisCO

 The 1st cpd formed is Oxaloacetic acid (OAA)

 OAA is converted into either Malate or Asparate

 Malate is decarboxylated in to Pyruvate & CO2

 Pyruvate moves to mesophyll cell to regenerate PEP

 CO2 is fixed by Calvin cycle in bundle sheath cell

30
 C4 plants can carry out photosynthesis at low CO2 concentration

b/c PEPCase have greater CO2 affinity than Rubisco.

 C4 plants have no photorespiration problem by having spacial d/ce

 separating CO2 fixation stage from the Calvin cycle in space .

Examples include sugar cane, maize, sorghum, etc

31
 Cont’d
Photosynthesis: Dark Rxn

32
3-CAM Pathways/ Crassulacean Acid Metabolism cycle
 Close Have diurnal stomatal pattern

 At night CO2 is absorbed by PEP due to PEPcase

 The 1st cpd is OAA and converted to malic acid & stored in vacuole.

 Malic acid is decarboxylated into Pyruvate & CO2

 Pyruvate is used for regeneration of PEP

 CO2 enters to Calvin (C3 cycle) to synthesis sugar/glucose.

 CAM plants minimize photorespiration and save water by separating
these steps in time, between night and day, i.e.,
 they close their stomata during the day to prevent water loss
 they open their stomata at night to allow the entry of Co2
-> temporal d/ce
Examples cacti, pineapples, orchids, etc
33
Dark: Stomata opened Light: Stomata closed

34
Key points:
• Photorespiration is a wasteful pathway that occurs when
the Calvin cycle enzyme rubisco acts on oxygen rather than
carbon dioxide.
• The majority of plants are C3 plants, which have no special
features to combat photorespiration.
• C4plants minimize photorespiration by separating
initialCO2 fixation and the Calvin cycle in space,
performing these steps in different cell types.
->spacial d/ce
• Crassulacean Acid Metabolism (CAM) plants minimize
photorespiration and save water by separating these steps in
time, between night and day.
->temporal d/ce 35
Factors Affecting Photosynthesis
The limiting factors that affect the rate of photosynthesis
can be categorized into internal and external groups

Internal External
 chlorophyll content  Rate of O2:CO2
 protoplasmic factors  light intensity
 nutrition (macro/micro)
 Temperature

 Water availability
36
Internal factors
Chlorophyll content
The actual rate of photosynthesis increases as chloro- phyll
content increases.
 About 20 to 100 Chloroplasts / Mesophyll Cell in Leaves

 Rate of Photosynthesis

 slow in young leaves

 Optimum in mature leaves

Light energy→ Chemical energy

37
Nutrition
 Nutrient affects photosynthesis activities

 Mg & Fe influence chlorophyll

 Cu influence co-factor enzymes

 Look the effect of Mg2+

38
 External factors
Ratio of O2 : CO2 in cells
 If ↓O2 but ↑ CO2, Normal photosynthesis i.e., Calvin

Cycle Dominates

 If ↑O2 and ↓CO2, Photorespiration Dominates

E.g., C3 vs C4: [RuBisCO vs PEPcase]

39
 ↑[CO2] can ↑rates of Photosynthesis

 eventually plateaus at maximum rate of fixation

Maxrate fixn
Rate of photosynthesis

↑rate fixn Maxrate fixn

CO2 conc.
40
Temperature
 Within physio Trange, ↑Temp will ↑ rate of PS
Below Normal Ranges, PS Slows or Stops

 Cell sap slows moving

 Cells may freeze

 Change protein & membrane structure

Above Normal Ranges

 Proteins may Change Shape

 Membranes become permeable

 Possible Cell Death

41
 Light
 PS carried out under various EM spectrum (VIBGYOR).

 Bacteria can photosynthesize under IR (λ = 900nm)

 Light affects photosynthesis in three aspects

 Light intensity

 Light quality

 Duration of light

42
Light intensity

 Leaf morphology, anatomy …

 Of the total quantity of incident light:
☼ Reflected,
☼ transmitted &
☼ absorbed
 Only 2% of Visible light is used

Light quality
 Chl absorbs light in Blue (λ = 450nm) & Red (λ = 660nm)
 These are the Photosynthetic Wavelengths of Light
 Called Photosynthetically Active Radiation (PAR)
43
 Cont’d
Factors Affecting

44
Duration of light

 Plants need sufficient length of light period to fix enough

carbons for normal growth

Controversies
 Quantum yield high exposed to continuous daylight of 10 -
12hrs

 Continuous light is damaging to photosynthesis

45
4.2. Respiration
What is Plant Respiration?
plants do not have any specialized organs like lungs to
breathe rather they respire.
Plants respire with the help of lenticels and stomata which
carry out the function of the gaseous exchange.
• The method by which cells get chemical energy by the
consumption of oxygen and the liberating of carbon dioxide
is called respiration.
• In order to carry on respiration, plant cells require oxygen
and a means of disposing of carbon dioxide just as animal
cells do.
• In plants, every part such as root, stem executes respiration
as plants do not possess any particular organs like animals
for the exchange of gases.
46
• All living organisms respire in order to release energy from
glucose and make it available in the form of ATP for chemical,
osmotic and other work.
• Plants respire in the normal way using glycolysis, Krebs cycle,
oxidative phosphorylation , etc.
• Plants need energy to take in mineral salts from the soil where
they are present in very low concentration - this needs work
(energy) to concentrate the mineral inside the plant.
• Plants growing in waterlogged soils (which are short of oxygen)
cannot respire in their roots and soon show the symptoms of
shortage of minerals (like yellow leaves).
• Moving sugars around the plant seems to require energy as
dead phloem cells do not transport sugars.
• Complex chemicals (like proteins) need energy to make them
from simple chemicals - again the plants need a supply of
energy to do this. 47
4.2.1. Glycolysis
• Glycolysis- is the process in which one glucose molecule is
broken down to form two molecules of pyruvic acid
(pyruvate).
is the first stages of respiration
Is anaerobic respiration
 It does not require oxygen.
 It breaks down one glucose molecule in to two pyruvate molecules.
 It Produces two net ATP molecules (4 gross ATP).
 It Produces two NADH molecules
 It takes place in cytoplasm
 Glycolysis is found in animals, plants and microorganism.
 Glycolysis involves ten enzymatic reactions.  ( See the figure below )
 This pathway is used by anaerobic as well as aerobic organisms.

48
49
 4.2.2. Krebs cycle and Electron Transport Chain
Kreb’s cycle
-the Kreb‘s cycle also called The Tricarboxylic Acid (TCA) cycle or Citric
acid cycle
-is aerobic respiration stage
-is considered as central pathway of aerobic metabolism,
Steps of the Krebs cycle
-acetyl CoA combines with oxaloacetic acid to form citric acid.
-takes place in mitochondrial matrix
*In one turn of TCA cycle:
– 2molecules of carbon dioxide are formed
– 1 ATP is formed,
– 3 NADH molecule s are formed
– 1 FADH is formed .
*In the complete (two turns) of TCA:
– 4carbon dioxide molecules are released,
– 2ATP molecules are formed
– 6NADH are produced
– 2FADH2 are produced
50
51
Electron Transport Chain
• The electron transport chain is a series of four protein
complexes that couple redox reactions, creating an
electrochemical gradient that leads to the creation of ATP
in a complete system named oxidative phosphorylation.
• In aerobic respiration, electron transport is the final step in
the break-down of glucose.
• It is also the point at which most of the ATP is produced.
• high-energy electrons and hydrogen ions from NADH and
FADH2 produced in the Krebs cycle are used to convert ADP
to ATP.

52
• An electron transport chain (ETC)
is a series of protein complexes and other molecules
that transfer electrons from electron donors to electron
acceptors via redox reactions (both reduction and oxidation
occurring simultaneously) and couples this electron transfer
with the transfer of protons (H+ ions) across a membrane.

• Many of the enzymes in the electron transport chain are

membrane-bound.
• The flow of electrons through the electron transport chain is
an exergonic process.
• The energy from the redox reactions creates
an electrochemical proton gradient that drives the synthesis
of adenosine triphosphate (ATP).
• In aerobic respiration, the flow of electrons terminates with
molecular oxygen as the final electron acceptor that provides
most of the energy.
53
• The H ions are pumped into the mitochondrial matrix across
the inner mitochondrial membrane.
• H ions then diffuse down their concentration gradient back
across the membrane and into the matrix through ATP
synthase molecules in chemiosmosis.
• Electron transport and chemiosmosis in cellular respiration
are similar to these processes in photosynthesis.
• Oxygen is the final electron acceptor in the electron
transport system in cellular respiration.
• Protons and electrons are transferred to oxygen to form
water.

54
55
• The electron transport chain releases the energy stored
within the reduced hydrogen carriers in order to synthesise
ATP
• This is called oxidative phosphorylation, as the energy to
synthesise ATP is derived from the oxidation of hydrogen
carriers
Oxidative phosphorylation occurs over a number of distinct
steps:
• Proton pumps create an electrochemical gradient (proton
motive force)
• ATP synthase uses the subsequent diffusion of protons
(chemiosmosis) to synthesise ATP
• Oxygen accepts electrons and protons to form water

56
 Summary
Electron Transport Chain
- occurs in mitochondrial cristae
-is aerobic stages of respiration
-is the final step in the break-down of glucose
-most of the ATP is produced here fromNADHandFADH2.
-H+ ions then diffuse down their concentration gradient
back across the membrane and into the matrix through
ATP synthase molecules in chemiosmosis.
-Oxygen is the final electron acceptor in ETC.
- Protons and electrons are transferred to oxygen to form
water.

57
5.Translocation
in the phloem

58
 Phloem anatomy
Phloem is much complex tissue than xylem, b/c
xylem is composed of vessels, tracheids, xylem fibres and xylem
parenchyma.
Phloem is composed of:
sieve tubes,
companion cells,
phloem fibres and
phloem parenchyma.
The main components are
 Sieve tubes /Sieve tube elements
 Companion cells

The sieve tube members are living cells (which do not contain a nucleus) that are
responsible for transporting carbohydrates throughout the plant.

The main functions of sieve tube members include:

maintaining cells and
transporting necessary molecules with the help of companion cells.
59
• The companion cells are specialized parenchyma cells in the phloem
tissues of the angiosperms.

The main function of the sieve tube is the transport of carbohydrates,

primarily sucrose, in the plant.

The interface of the tubes contains pores which help in conduction.

Each sieve tube element is normally associated with one or more
nucleated companion cells, to which they are connected by
plasmodesmata.

Function of companion cell is to load sugar and amino acids into sieve
elements.

These cells use transmembrane proteins to take up these molecules by

active transport.

i.e., to provide energy to the sieve tube element during translocation 60

Phloem anatomy

Companion cell
1o Phloem
1o Xylem
61
The distinguishing feature of phloem tissue is the conducting
cell called the sieve element/sieve tube.
Sieve tubes
 Sieve elements:
 arranged longitudinally end to end
 have protoplasts (p-protein=phloem protein)
 have no nucleus and
 have reduced numbers of organelles to maximize space
for the translocation of materials.
 have thick and rigid cell walls to withstand the hydrostatic
pressures which facilitate flow
 sieve plates: separated by perforated end wall
-have thin & extensible cellulose wall, i.e.
Porous cross walls
-narrow as compared to xylem vessels 62
• The perforations in sieve plates allow water and dissolved
organic solutes to flow along the sieve tube.
• The sieve plates are lined with callose.

63
• the pores of the sieve area are relatively large and are
found grouped in a specific area, they are known as sieve
plates.
• Sieve plates are typically found in the end walls of sieve-
tube members and provide a high degree of protoplasmic
continuity between consecutive sieve-tube members.
• Additional pores are found in sieve areas located in lateral
walls.
• Sieve plates are the connecting and transport tissue in
plants.
• Sieve plates allow the food to pass through the phloem
tubes.
• The tiny pores present on these tubes helps in the
transport and absorption of food particles.
• Thes have long and elongated structures that connect the
roots and all other parts of plants. 64
Characteristics of sieve elements in seed plants
 Sieve tube in Angiosperms:
 Some sieve areas differentiated into plates;

 individual elements are joined into a sieve tube

 Sieve plate pores are open channels,

 P-protein is present in all dicots &many monocots

 Companion cells are source of ATP for sieve tubes

b/c they have organelles

65
 Sieve cells in Gymnosperms:
 No sieve plates; all sieve areas are similar

 No p-protein,

 No companion cells, but, Albuminous cells

 Pores in sieve areas appear blocked with membranes

66
Companion Cells
 are parenchymal type of cells

 oriented longitudinally among sieve tube

 are living cells

 intimate relation with sieve tubes in function

 energy providing to sieve tubes b/c Each sieve tube element

is associated with one or more companion cells.

 Participate in loading & unloading

67
Phloem anatomy

68
Mechanism of phloem translocation
What is transported in phloem?
->the chemical composition of phloem exudate is highly variable.
It depends on:
-the species,
-age, and
-physiological condition of the tissue sampled.
For e.g., an analysis of phloem exudate from stems of actively growing castor bean
(Ricinus communis) shows that the exudate contains:
-> sugars,
-> protein, amino acids,
-> the organic acid malate,
-> avariety of inorganic anions (phosphate,sulphate, and chloride)
-> cations (the predominant is potassium).
-> Some plant hormones (auxin, cytokinin, and gibberellin) but at very low
concentrations.
The principal constituent of phloem exudate in most species is sugar.

The most common transported sugar is sucrose.

69
70
Phloem transports photo assimilates:
-from the site of formation (source-eg. mesophyll cell of leaf
-To the site of consumption or storage (sinks).
Sinks include:
-> areas of active growth (apical and lateral meristems,
developing leaves, flowers, seeds, and fruits)
or areas of sugar storage (roots, tubers, and bulbs).
Storage locations can be either a source or a sink, depending
on the plant‘s stage of development and the season.

• Translocation is the movement of sucrose around a plant

from the source to the sink.
71
Experimental evidences
Several experimental evidence supports the
translocation of phloem sap(sucrose, other organic
solutes, hormones, minerals ,etc) by phloem.
 Ringing /girdling experiment

 Blocking of sieve pores

 Chemical analysis of phloem sap

 Stem feeding aphids

72
1- Ringing experiment :

-ringing below photosynthetic leaf

results in swelling above the ring
(A).

-ringing above photosynthetic

leaf results in swelling below
the ring (B).

** this indicates translocation

is a task of phloem
73
2- Blocking of sieve pores:
phloem tissue is composed of:
 sieve tubes(contains sieve pores)
 companion cells
 phloem parenchyma
Therefore, blocking sieve pores blocks translocation
3-Phloem sap analysis by using stem feeding aphids
Aphid uses stylet to tap into phloem.
The aphid body cut from stylet after stylet inserted into
phloem.
When the stylets are withdrawn the stylet sheath remains in
the tissues of the plant and indicates the exact passage taken by
the stylets.
Stylets placed at different parts of the plant can show rate of
movement of phloem sap. 74
Aphids feed on the sap of plants through the plant's phloem vessels.
Phloem is a type of transport tissue that moves sugars throughout the
plant to where they're needed. This is the main sap aphids are after,
but they also feed on xylem sap.

Aphids feed directly on phloem sap, by inserting their specialised

mouthparts into the plant's phloem and ingesting the sap by sucking.

Thus by collecting honeydew from aphids feeding on various parts of

plants one could determine which plant hormones are being
translocated, and also any changes in the level of these hormones with
the state of growth of the plant.
75
4-Radioactive tracer experiment
• If radioactive traces like 14CO2 or (14C) sucrose is fed to
leaves and chased,
i.e. followed with time, it is possible to follow the
pathway through which the photosynthate or sucrose is
moving.
Mechanism of phloem translocation can be explained
by the following hypothesis.
 Diffusion hypothesis:
 Activated diffusion
 Protoplasmic streaming
 Transcellular streaming
 Modified pressure flow 76
Diffusion hypothesis:

Phloem sap translocation takes place by principles of diffusion

• During photosynthesis, the cell sap concentration in
mesophyll cells is high and as a result high osmotic
pressure exists in them.
• Due to high osmotic concentration, mesophyll cells absorb
water from the adjacent tissues.
• This creates a high turgor pressure.
• Mesophyll cells are connected to sieve tubes through small
pores, known as plasmodesmata.
• This makes a continuous channel through which the solutes
flow in bulk under the osmotic influence.
• i.e., If the concentration at the source is high and low at the
sink, then phloem sap can move.
77
• Diffusion hypothesis believes that translocation is
fundamentally simple diffusion and translocation will take place only
if there is a concentration gradient between the supply end and
consumption end.
• If there is no concentration gradient there is no translocation.
• The rate of translocation will be greater when the concentration of the
solutes in the supply end is also greater.
• The only merit of the theory is that it explains the simultaneous flow
of organic compounds in opposite directions.
• But this theory is also not accepted because translocation of solute is a
rapid process whereas diffusion is a very slow process.

• So it cannot account for the rapid movement of food materials.

78
Cyclosis (cytoplasmic streaming)
• According to this, moving protoplasm carries the solutes within the
sieve elements and the protoplasmic fluid moves from cell to cell
through large protoplasmic connections across the sieve plates.
• A circular movement or Cyclosis of living protoplasm has been
observed in many different plant cells.
• On the basis that sieve tubes are living system, so that cytoplasmic
streaming /cyclosis in sieve elements operates in the sieve tubes.
• Thus substance would be carried downward or upward
 i.e., is the flow of the cytoplasm inside the cell, driven by forces
from the cytoskeleton.

 However, the presence of cyclosis even in the undeveloped

sieve tubes and also the occurrence of the sieve does not fully-
support this hypothesis
79
The evidences in support of this theory are:
1. Streaming of granular substances is observed in all living cells.
2. Streaming will allow a rapid transport than that of diffusion.
3. The solutes will be easily carried from one place to another by the
streaming protoplasm
4. Protoplasmic connections are observed between adjacent sieve
tubes through the pores present in the sieve plate
5. A recent evidence in favour of this theory comes from the works of
Thaine (1967).

The major objections against this theory are:

i) The cytoplasmic streaming has never been observed in
mature sieve elements
ii) The rate-of streaming cannot account for the rapid
translocation of solutes.

80
 Pressure-flow hypothesis/ model
 states that ―a flow of solution in the sieve elements is driven
by an osmotically generated pressure gradient b/n source&sink.
 This is the most commonly accepted hypothesis to explain the
movement of sugars in phloem.
*a high concentration of sugar at the source creates a low solute
potential (Ψs), which draws water into the phloem from the
adjacent xylem.
*This creates a high pressure potential (Ψp), or high turgor
pressure, in the phloem.
*The high turgor pressure drives movement of phloem sap by
―bulk flow‖ from source to sink, where the sugars are rapidly
removed from the phloem at the sink.
*Removal of the sugar increases the Ψs, which causes water to
leave the phloem and return to the xylem, decreasing Ψp.
81
In source tissue, phloem loading leads to a buildup of sugars
 Makes low (-ve) solute potential
 Causes a steep drop in water potential
 In response to this new water potential gradient, water
enters sieve elements from xylem
In sink tissue, phloem unloading leads to lower sugar concentration
 Makes a higher (+ve) solute potential
 Water potential increases
 Water leaves phloem and enters sink sieve elements and
xylem
• Thus phloem turgor pressure decreases
N.B Translocation stops if the phloem tissue is killed.
82
 Cont’d
Mechanism of translocation

83
 Phloem Loading: Where do the solutes come from?

 Triose phosphate – formed from photosynthesis during the day is

moved from chloroplast to cytosol
 At night, this compound, together with glucose from stored starch, is
converted to sucrose.
 Both these steps occur in a mesophyll cell
 Sucrose then moves from the mesophyll cell via the smallest veins in
the leaf to near the sieve elements.
 The transfer of sugars (photosynthetic) from mesophyll cells to sieve tube
elements in the leaf is called as phloem loading.
 the transfer of sugars (photosynthetic) from sieve tube elements to the
receiver cells of consumption end (i.e., sink organs) is called as phloem
unloading.

84
 In a process called sieve element loading, sugars are transported
into the sieve elements and companion complex cells.
 Sugars become more concentrated in sieve elements and companion
cells than in mesophyll cells.
 Once in the sieve element /companion cell complex sugars are
transported away from the source tissue – called export.

85
86
 Symplastic phloem loading

 Dependant on species that transport sugars other than

sucrose
 Requires the presence of open plasmodesmata between different cells
in the pathway
 Dependant on plant species with intermediary companion cells
• Sucrose, synthesized in mesophyll, diffuses into intermediary cells
• Here Raffinose is synthesized.
• Due to larger size, can NOT diffuse back into the mesophyll cell.
• Raffinose and sucrose are able to diffuse into sieve element

87
What is apoplastic and symplastic pathways?
Apoplast Pathways:
Water goes from root hairs to xylem entirely via the cell
wall, without crossing any membranes, via the apoplast
pathway.
Symplast Pathways:
Water moves from cell to cell through protoplasm through
the symplast route, which is aided by plasmodesmata.

88
 Phloem unloading
Three steps
(1) Sieve element unloading:
– Transported sugars leave the sieve elements of sink tissue
(2) Short distance transport:
– After sieve element unloading, sugars transported to cells
in the sink by means of a short distance pathway
(3) storage and metabolism:
– Sugars are stored or metabolized in sink cells

89
 Phloem unloading
 Can also occur by symplastic or apoplastic pathways
 Varies greatly from growing vegetative organs (root tips and young
leaves) to storage tissue (roots and stems) to reproductive organs
 Symplastic:
 Appears to be a completely symplastic pathway in young dicot leaves
 Again, moves through open plasmodesmata

90
 Phloem unloading
Apoplastic: three types
(1) transport from the sieve element-companion cell complex
to successive sink cells, occurs in the apoplast. [see fig B
below].
Once sugars are taken back into the symplast of adjoining
cells transport is symplastic

91
(2) [Fig A below ] involves an apoplastic step close to the
sieve element companion cell.
(3) [B] involves an apoplastic step further from the sieve
element companion cell
• Both involve movement through the plant cell wall

92
 Patterns of Phloem translocation
Translocation of Phloem Sap
Phloem translocation is defined with respect to source  sink.
Phloem sap is an aqueous solution of
 sucrose
 minerals,
 amino acids
 Hormones
Sugar source
 is a plant organ:
 site of photosynthesis (leaves)
 site of breakdown of starch
Sugar sink
 is a plant organ: non-photosynthesis
 net consumer (roots, shoots , young leaves ,
 storage organ (tubers, bulbs, fruits, seeds)
 Factor affecting Phloem Translocation
Temperature
 The lowest To retards translocation

 [20 – 30oC] The highest peak of translocation rate

Light
 Low light intensity inhibits root & shoot growth

Less demand of food  Poor translocation

Oxygen availability
 poor oxygen availability retards translocation
 since translocation is energy demanding process

Metabolic effect
94
 factors affecting rate of respiration affect rate of translocation
# Why plants transport sugars as sucrose and not glucose?
It is interesting to speculate on why sucrose is the preferred vehicle for
long-distance translocation of photoassimilate.
* The possibility is that:
-sucrose is a disaccharide composed of glucose and fructose
-is non-reducing sugars.
-nonreactive in alkaline solution b/c the acetal link b/n the
subunits is stable.
-has a low viscosity even at high concentrations and has no
reducing end and
-is thus considered more inert than glucose, which is the major
transport form in animals.
* all monosaccharides are reducing sugars & have a free aldehyde or ketone
group that is capable of reducing mild oxidizing agents.
*During photosynthesis, glucose is produced.
Before it is transported, glucose is converted to sucrose.
Finally it will be stored in the form of starch.
95
 6. Introduction to Nitrogen Metabolism
Nitrogen is a naturally occurring element that is essential for growth and
reproduction in both plants and animals.

It is found in:

-amino acids that make up proteins,

-nucleic acids (DNA&RNA)hereditary material, life's blue print for all cells.

-ATP—the universal energy source

-the molecule of chlorophyll( Chla (C55H72MgN4O5, Chlb (C55H70MgN4O6)

-any other organic(Vitamins) and inorganic compounds.

96
 Nitrogen metabolism

is defined as the conversion of molecular nitrogen into fixed form of

nitrogen to make it available for absorption by plants.

i.e., N2 → →NO3- or NH3

 The process of converting inorganic nitrogen cpds into

organic form is important to form:
e.g., -amino acids( protein),
-nucleic acid,
-hormone,
-enzymes( co-factors, co-enzymes)
97
Nitrogen Fixation
 Nitrogen is one of the most abundant elements on earth.

 Nitrogen moves slowly through the cycle and occurs in 3 main

reservoirs such as the atmosphere, soils, living organisms, and
oceans. Most of the nitrogen on Earth is in the atmosphere.

98
 About 80% of the atmosphere is composed of N2
 N2 fixed in nature is ~ 230 x 106 Tm/ annum
 13 % is fixed by electric storms e.g., nitric acid
 87 % is result of biological fixation
e.g., 80% is fixed via symbiotic associations
20% is fixed via free living organisms
The main steps in the N-cycle include:
i. Nitrogen fixation: N2 → → → NH3 [bacteria]
ii. Decomposition/Ammonification: releasing of excess NH3 & NH4+
from dead plants & animals , including their waste products like urine, fece
iii. Nitrification: NH3 & NH4+ → →NO2- → → NO3-
iv. Denitrification: NO2- & NO3- → → N2 …..[bacteria]

99
100
Sources of nitrogen

The major sources of nitrogen are atmosphere & soil.

The major sources of Nitrogen in the soil are:

i. Nitrate nitrogen
ii. Ammonia nitrogen
iii. Organic nitrogen
iv. Molecular nitrogen

*Plants get N2 mainly in the form of nitrate from soil.

101
I-Nitrate nitrogen:
 decomposition of OM  NO3- in the soil
NO3-  NH4+ within living plant cells,
 various enzymes are involved in this process
e.g, nitrate reductase, nitrite reductase,
nitric oxide reductase, hyponitrite reductase,
hydroxylamine reductase
II-Ammonia nitrogen:
 NH+4 is a cation and so held tightly to soil particles

 nitrification process by soil bacteria: (HNO3)

NH+4 → → NO2-  → HNO3
 NH3 is made available to plants by changing NO2- into NO3-
 urea is also a good source of NH3
NH2CONH2 + H2O → 2NH3 + CO2 102
III-Organic nitrogen:
 obtained from decaying organic materials

 not directly absorbed by plants except for urea

 organic molecules in the soil particle first break down to

amino acids and then followed by oxidation into
ammonia and organic acids

 the ammonia thus released, immediately gets oxidized

into NO-3, which on absorption by plants gets converted
to ammonia
103
Nitrogen fixation can be classified as:
Nitrogen fixation

Artificial fixation Natural fixation

Biological Non-biological

Symbiotic Non-symbiotic 104

Artificial fixation
 Haber-Bosch: under specially industrial condition at high temperature and

pressure atmospheric nitrogen chemically combines with

hydrogen gas to form ammonia.

N2 + 3H2 2NH3
Natural fixation

 Can be classified as:

 Non-biological N-fixation (physical )

 Biological N-fixation (microorganisms)

105
Non-biological N-fixation (physical )
 Carried out by physical factors without living organisms
 Occurs during rainy season:
lightning, thunders, stores, atmospheric pollution or electrical fixation.
N2 + O2 → → 2NO [lightning]

2NO + O2 → → 2NO2 [oxidation]

2NO2 + rain water → HNO2

2NO3 + rain water → HNO3

 Alkaline radical: Ca+2 & K+ react Ca(NO3)2 or KNO3

Biological N-fixation ( by microorganisms)

 requires participation of d/t organisms & enzymes

 mediated by:
i) Non-symbiotic microorganisms
ii) Symbiotic microorganisms 106
 Non-symbiotic N fixation can be carried out by
free living bacteria in soil like:
 Blue green algae,
Clostridium,
 Azotobacter,
Nostoc
 Symbiotic N fixation is carried out by symbiotic
bacteria that live symbiotically with the root of
leguminous plant like
 Rhizobium 107
Assimilation of Ammonia & Nitrate

Major steps in which nitrogen from the soil is assimilated in to the plants
system are

i. nitrate reduction,

ii. nitrite reduction

iii. reduction of hydroxylamine

Nitrate reduction:

According to Evans and Nason (1954),

N2 → → → NO3- [nitrogen reductase ]

H+ is furnished from NAD which is used for reduction of nitrogen.
108
Nitrite reduction:

 nitrate is further reduced to nitrite

 es- come from photochemical reaction

NADH + H+ + NO3- → → →NO2- + H20 + NAD+

Reduction of hydroxylamine
 hydroxylamine to ammonia

NH2OH + NADH + H+ →→ → → → NH3 + NAD+ + H2O

by hydroxylamine reductase

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7. Growth & development of plant

The entire process of growth and development in plants is

carried out by the specialized cells – meristems.

110
Growth: What is growth?
Growth can be defined as an irreversible permanent increase in size,
weight, length, volume & Surface area of an organ or its parts or
even of an individual cell.
Growth is accompanied by metabolic processes (both anabolic and catabolic) that occur
at the expense of energy. For example, expansion of a leaf is growth.
Growth describes a quantitative change in a plant or a plant part.
• Plant growth is unique because plants retain the capacity for unlimited
growth throughout their life.=>indeterminate growth
This ability of the plants is due to the presence of meristems.
These meristems have the capacity to divide and self-perpetuate.
 Growth takes place at specific regions in the plant body.
These growth regions are:
i. 10 growth regions- are apices of shoot & root
ii. 20 growth regions- are cork and vascular cambium
Summary of plant growth
1-Primary growthprimary tissue in lengthby SAM
&RAM
primary tissues :-Tissues that are derived directly from the root
and shoot apical meristems.

The primary tissue of roots and shoots contains a central core

of vascular, or conducting elements.

 The root apical meristem (RAM) is a cluster of dividing

cells located at the tip of the root just behind the root cap.
 The shoot apical meristem generates the aerial organs of the
plant such as: leaves, branches, and floral parts.
112
2-secondary growth secondary tissue in diameter/girth
By the vascular cambium
Secondary tissues: tissues laid down by the vascular cambium,
so the vascular cambium is responsible for secondary growth.

• The stems and roots of woody plants, however, grow in

diameter as well.

• An increase in diameter results from the activity of a

meristem called the vascular cambium.

The vascular cambium develops between xylem and phloem

and produces new xylem toward the inside and
new phloem toward the outside.
113
Development:
It is defined as all the changes that an organism goes through
during its life cycle, right from seed germination to senescence.
i.e., the transition from embryo to seedling,
from a leaf primordium to a fully expanded leaf,
or from the production of vegetative organs to the
production of floral structures.
It is a qualitative change occurring in a plant
*Development is the sum of growth, differentiation &
morphogenesis (the acquisition of form and structure).
It is an umbrella term, referring to the sum of all of the
changes that a cell, tissue, organ, or organism
goes through in its life cycle.
114
*Both growth & development are regulated by internal & external factors

i-Internal factors e.g., genes, plant hormones, photosynthesis

Ii-External factors e.g., light, air, water, mineral, gravity, temperature, day length

Growth Kinetics :

Growth kinetics is an autocatalytic reaction which implies that the rate

of growth is directly proportional to the concentration of cell.

 Growth is not a spontaneous process

 Meristematic cell in growth regions has to pass three phases

i- Cell formation

ii-Cell elongation

iii-Cell differentiation
115
Cell formation:
 basic process in growth
 increase the No of cells in mitotic division in all
somatic cells
 rate of division depends on the
 type of cells
 type of species
 other conditions (physical, chemical)

116
Cell elongation:

 increase in:
 cell size and volume: cytoplasmic content
 number of organelles
 adding organic material
 addition of water
 each cell grows to the limit: the cell divides before it
reaches the limit

117
Cell differentiation:

 is the process in which the cells specialize into morphologically

and physiologically different cells to perform particular function

 the cell undergoes change in form in order to perform

specialized functions.

 the cells start to differentiate from each other in its structure

and chemical composition

e.g., meristematic vascular cells

 the meristematic vascular cells differentiate into: Xylem and phloem

118
 Dedifferentiation-mature plant cells can be stimulated under certain
conditions to divide and differentiate again,
 This usually occurs when tissues are wounded,
as when branches break or
leaves are damaged by insects
plant repairs itself by dedifferentiating parenchyma cells in the
vicinity of the wound, making cells like those injured or else
physiologically similar cells.
• Plants differ from animals in their manner of growth.
• As young animals mature, all parts of their bodies grow until they
reach a genetically determined size for each species.
• Plant growth, on the other hand, continues throughout the life span of
the plant and is restricted to certain meristematic tissue regions only.

119
Plant Growth Analysis

120
According to their physiological basis Sigmoid (growth)
curve has five phases
 Phase-1------Lag phase
 Phase-2------Exponential phase
 Phase-3------Linear phase
 Phase-4------Declining phase
 Phase-5------Negative phase

121
Fertilizatio

Germination
Zygote Embryo Seed

Reproductive stage Seedling

Phase 1: Lag phase
-occurs just after germination period
-foliage canopy is very small & trap little light

-low photosynthetic rate

-dry weight increases very slowly

-slope of the curve is very low

-The growth is slow at this stage.

The lag phase is the adaptation phase for the organism where they

acclimatizes themselves to the new environmental conditions provided.

122
plant uses the nutrients stored in seed and begins to develop
 Epicotyl shoot apical meristem
 Radicle an immature root system

Phase 2: Exponential phase

-foliage canopy grows rapidly

-light trapping efficiency increases rapidly

-increase in the photosynthetic rate

- total dry weight increases exponentially

-slope of the curve increases rapidly

-high photosynthetic tissues are produced

123
Phase 3: Linear phase
 total dry weight increases at a constant rate.
♣ slope of the curve is constant
 foliage canopy continues to grow
♣ light interception continues to increase
♣ increase photosynthesis process
♣ increases non photosynthetic tissues
♣ continuous increase in dry weight
♣ In photosynthetic tissue-> photosynthesis + respiration process
♣ In non photosynthetic tissues-> respiration process
♣ The net result of: Dry weight = photosynthesis – 2xrespiration
process

124
Phase 4: Declining phase
 total dry weight become decreases

♣ slope of the curve decrease gradually until it

becomes zero at the end of the phase.

The decrement for dry weight occurs due to two reasons

i. increment in non photosynthetic tissues
ii. mutual shading
The net result of:
Dry weight = photosynthesis – respiration process

125
Phase 5: Negative phase
 total dry weight decreases with time
♣ slope of the curve is negative

This phase occurs due to several reasons

 Stopping of leaf growth [no new leaves]
 Leaf ageing [photosynthetic rate ↓]
 Leaf shedding [Senescence]
 Senescence is an aging process characterized by:
increased respiration,
declining photosynthesis,
and an orderly disassembly of macromolecules.
126
Factors Affecting Plant Growth
• Temperature: growth is accelerated with the increase inTo
• Light: intensity, duration and quality of light influences
many physiological processes occurring in a plant.
• Water: is an essential factor for plant growth.
• plants grow well in sufficient amount of water.
• plants even respond to the scarcity of water.
• Soil Nutrients:
Plants require an adequate amount of nutrients for proper growth.
The quality and quantity of nutrients affect plant growth.
• Plant Growth Regulators:
plant growth can be regulated by auxin, cytokinin, gibberellins, etc

127
 Plant Growth Regulator Substances (Hormones)

General characteristics of hormones

 are internal factors (chemical substances)

 are either natural or synthetic chemicals

 have either stimulating, promoting, enhancing,

retarding, inhibiting or suppressing effect

 are produced & physiological active in minute amount

 i.e., Hormones are the chemical messengers that

enable cells to communicate with one another.
Five major kinds of plant hormones are well known
 Auxins

 Cytokinins

 Gibberelins

 Ethylene

 Abscisic acid

Auxins:
 first to be identified in plants
 Either natural or synthetic chemicals
129
 Auxins tend to migrate away from light
B/c the phototropins/photoreceptor proteins /flavoproteins
are more active on the side with light, causing the
auxin to flow down the shady side by mediate
phototropism responses in higher plants.
Photoreceptors are proteins that plants use to correctly
interpret light and activate processes to regulate
downstream signaling pathways which influence plant
physiology, growth, and development.

130
 the most widely distributed natural auxin is Indole-3-
Acetic Acid, IAA (C10H9O2N)
 IAA is synthesized in meristems, young leaves, and

developing fruits and seeds(embryo).

• Transported unidirectionally towards the bases

• Although virtually all plant tissues appear to be capable
of producing low levels of IAA, shoot apical
meristems and young leaves are the primary sites of
auxin synthesis.

131
 Besides IAA, Indolebutyric acid (IBA),
Indole -3- acetaldehyde (IAAID),
Indole -3- ethanol (IETOH) and
Indole pyruvic acid (IPYA)
Indole Propanoic Acid (IPA),
are synthetically occurring Auxins Hormones

132
Roles of auxin:
 preventing leaf fall – by preventing formation of
abscission layer /i.e., delays the onset of leaf abscission
 xylem maturation and differentiation
 influences nearly every stage of a plant's life cycle from
germination to senescence.
 Auxin promotes fruit development
 promotes the formation of lateral and adventitious roots
 regulates apical dominance
 promote growth in stems, while inhibiting growth in root,
i.e, ↓conc. Auxins------promotes root formation
↑conc. Auxins------inhibits root formation

133
Agricultural application of auxins:
 promoting germination by reducing dormancy
 rooting – soaking stem cuttings of roses produces
adventitious roots if appropriate conc. is used
 preventing premature drop of fruits/Improve fruiting
 Serve as weed control herbicide by 2-4 D (2-4-
dichlorophenoxyacetic acid)
 suppression of lateral bud growth
 auxins promote cytokinins formation

134
Gibberelins:
 investigated from rice affecting fungus known as Gibberella
fujikuroi
 G. fujikuroi results ‗bakanae‘ (foolish seedling) disease of rice
 isolated chemical named Gibberellins.
 Common types of Gibberellins are
 GA1 (C19H24O6)
 GA2 (C19H26O6)
 GA3 (C19H22O6)
 GA4 (C19H24O5)
 GA3 is reported from large number of plants

135
Roles of Gibberellins:
 Breaking dormancy
 initiating seed germination
 Inhibiting root growth at high conc.
 Initiating Stem elongation
 Inhibiting flowering in fruit trees
 May result male sterility by suppressing growth of
androecium

136
Cytokinins:
 produced from component of Nucleic Acid
 Natural growth hormones found in d/t plants
 ‗Zeatin‘ is the most abundant naturally occurring free
cytokinin
• Cytokinins are synthesized in roots, developing embryos, young leaves
and fruits
• A major site of cytokinin biosynthesis in higher plants is the root.

137
Roles of Cytokinins:
 Increasing rate of cell division
 Enlargement of cells in leaves
 Formation of cambium
 Promoting germination by breaking dormancy
 Delay of senescence (aging & death of plants)
 Promoting translocation of solutes
 Promoting flowering
 Nucleic acid metabolism
 Protein synthesis
138
Ethylene:

Ethylene is a simple gaseous PGR.

It is synthesized in large amounts by tissues
undergoing senescence and ripening fruits.
Ethylene occurs in all plant organs – roots, stems,
leaves, bulbs, tubers, fruits, seeds, and so on

139
 ROLES OF ETHYLENE
 promoting fruits ripening
 move by diffusion

 Activation of enzymes: pyruvate decarboxylase

 Ethylene breaks seed and bud dormancy
 Inhibiting elongation of roots and stems
 Influences flowering in most plants
 Promoting changes that occur before leaf abscission
 Prevent Leaf and fruit abscission
 Release from dormancy

 It enhances the respiration rate during ripening of the

fruits.
140
Abscisic acid (ABA) :
 naturally occurring growth inhibitor hormone
 Abscission –refers to the shedding of leaves, flowers,
and fruits from the living plant.

141
Roles of ABA
 Causing seed dormancy
 Encourage leaf senescence
 Leaf abscission
 Influencing release of ethylene
 Inhibiting other hormones like GA and IAA
 Inhibition of shoot growth (suppresses bud growth)
 Stomatal functioning by its effect on guard cells related to k+
transport in response to water stress
 Signal for induction of drought, heat, cold and salt stress genes
 A typical effect of ABA on leaves is to reduce transpirational
142
8.2.1 Harmful interactions between plants, pathogens and herbivores

 Plants have developed various modes to protect themselves

against insect pests and pathogens.

1 -mechanical barriers,

2 -constitutive chemical defences,

3-direct as well as indirect inducible defences.

143
1. Mechanical barriers offer the first line of defence
against pests &pathogens for several species of plants that
include:

-surface structures(thorns, spines, prickles, and

trichomes),

-mineral crystals (silica crystals, phytoliths, and calcium

oxalate crystals raphides)

-thigmonastic (touch-induced) leaf movements in Mimosa

pudica.
144
THE END OF THE COURSE !

145