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MOI UNIVERSITY
SCHOOL OF SCIENCE AND AEROSPACE STUDIES
DEPARTRMENT OF BIOLOGICAL SCIENCES
BOT 313: PLANT PHYSIOLOGY, Lecture 3
Potential and turgor pressure.
Osmosis
Osmosis Definition
“Osmosis is a process by which the molecules of a solvent pass from a solution of low concentration to a solution
of high concentration through a semi-permeable membrane.”

Difference between turgor pressure and pressure potential.

Plant cells are very much like your own cells, except they are surrounded by a cell wall.
 This cell wall is part of what gives plants such a rigid and sturdy structure.
 Plant cells need a certain amount of pressure to make sure that the cell wall stays rigid.
 Pressure from fluid within the cell pushing against the cell wall is called turgor pressure.
 Presure ptential Symbol Ψ is the component of water potential due to the hydrostatic pressure that is
exerted on water in a cell.
 In turgid plant cells it usually has a positive value as the entry of water causes the protoplast to push
against the cell wall.
 In xylem cells there is a negative pressure potential, or tension, as a result of transpiration.
 Water at atmospheric pressure has a pressure potential of zero.

Osmosis
What is Osmosis?
 Osmosis is a passive process and happens without any expenditure of energy.
 It involves the movement of molecules from a region of higher concentration to lower concentration
until the concentrations become equal on either side of the membrane.
 Any solvent can undergo the process of osmosis including gases and supercritical liquids.
 Let us have a detailed look at the different types and effects of osmosis in detail.

Osmotic Solutions
There are three different types of solutions:
i. Isotonic Solution
ii. Hypertonic Solution
iii. Hypotonic Solution
 An isotonic solution is one that has the same concentration of solutes both inside and outside the cell.
 A hypertonic solution is one that has a higher solute concentration outside the cell than inside.
 A hypotonic solution is one that has a higher solute concentration inside the cell than outside.

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Types of Osmosis
Osmosis is of two types:
 Endosmosis– When a substance is placed in a hypotonic solution, the solvent molecules move inside
the cell and the cell becomes turgid or undergoes deplasmolysis. This is known as endosmosis.
 Exosmosis– When a substance is placed in a hypertonic solution, the solvent molecules move outside
the cell and the cell becomes flaccid or undergoes plasmolysis. This is known as exosmosis.

Effect of Osmosis on Cells

 Osmosis affects the cells differently.
 An animal cell will lyse when placed in a hypotonic solution compared to a plant cell.
 The plant cell has thick walls and requires more water.
 The cells will not burst when placed in a hypotonic solution.
 In fact, a hypotonic solution is ideal for a plant cell.
 An animal cell survives only in an isotonic solution.
 In an isotonic solution, the plant cells are no longer turgid and the leaves of the plant droop.
 The osmotic flow can be stopped or reversed, also called reverse osmosis, by exerting an external
pressure to the sides of the solute.
 The minimum pressure required to stop the solvent transfer is called the osmotic pressure.

Osmotic Pressure
 Osmotic pressure is the pressure required to stop water from diffusing through a membrane by
osmosis.
 It is determined by the concentration of the solute.
 Water diffuses into the area of higher concentration from the area of lower concentration.
 When the concentration of the substances in the two areas in contact is different, the substances will
diffuse until the concentration is uniform throughout.
 Osmotic pressure can be calculated using the equation:
Π=MRT
where Π denotes the osmotic pressure,
M is the molar concentration of the solute,
R is the gas constant,
T is the temperature

Significance of Osmosis
 Osmosis influences the transport of nutrients and the release of metabolic waste products.
 It is responsible for the absorption of water from the soil and conducting it to the upper parts of the
plant through the xylem.
 It stabilizes the internal environment of a living organism by maintaining the balance between water
and intercellular fluid levels.
 It maintains the turgidity of cells.
 It is a process by which plants maintain their water content despite the constant water loss due to
transpiration.
 This process controls the cell to cell diffusion of water.
 Osmosis induces cell turgor which regulates the movement of plants and plant parts.
 Osmosis also controls the dehiscence of fruits and sporangia.
 Higher osmotic pressure protects the plants against drought injury.

Examples of Osmosis
 Osmosis has a significant role to play in plants, animals and also in humans.
 In an animal cell, osmosis helps in absorbing water from the intestines to the blood.

Listed below are more examples of Osmosis.

 The absorption of water from the soil is due to osmosis.
 The plant roots have a higher concentration than the soil. Therefore, the water flows into the roots.

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 The guard cells of the plants are also affected by osmosis. When the plant cells are filled with water, the
guard cells swell up, and the stomata open.
 If a freshwater or saltwater fish is placed in the water with different salt concentrations, the fish dies
due to the entry or exit of water in the cells of the fish.
 Humans suffering from cholera are also affected by osmosis.
 The bacteria that overpopulate the intestines reverse the flow of absorption and do not allow water to
be absorbed by the intestines, which results in dehydration.
 When the fingers are placed in water for a longer period of time, they become pruney due to the flow
of water inside the cells.

Factors affecting the rate of Osmosis?

The factors affecting the rate of osmosis include:
1. Pressure.
2. Temperature.
3. Surface Area.
4. Water Potential.
5. Concentration gradient.

The significance of osmosis?

The biological importance of osmosis includes:
1. It is essential for the survival of a cell.
2. Osmosis plays a key role during the germination of seeds.
3. Involved in the movement of water molecules between the cell and cell organelles.
4. In plants, it is involved in the movement of water molecules from the soil into the root nodules.
5. The mechanism of stomata is mainly because of the response to the osmotic pressure of the guard cells
in relation to the epidermal cells.
The transport of water molecules from the region of higher concentration to the region of lower concentration
through a semipermeable membrane is called osmosis.

Importance of osmosis in the life of plants:

1. Osmosis helps in retaining the turgidity of the cell.

2. It plays an important role in the movement of stomata during transpiration.
3. The opening and closing of stomata are maintained by osmosis.
4. It helps in the movement of liquid through a biological membrane.
5. It plays a vital role in the growth of radicle and plumule at the time of germination of seeds.

Cell membrane
PERMEABILITY, FLUIDITY, HETEROGENEITY
 Cell membrane functions are determined by their molecular composition.
 Three main membrane features:
1. Semi-permeability,
2. Fluidity and
3. Lateral heterogeneity.

1. Permeability
 Membrane semi-permeability is a consequence of their inner hydrophobic environment generated by
the lipid fatty acid chains.
 This hydrophobic space is difficult to be crossed by molecules having net electric charge.
 Thus, by preventing free diffusion of some molecules, membranes can form compartments that keep
distinct internal and external environments, as well as an intracellular milieu different from that of the
extracellular space.
 However, permeability is selective, that is, not all molecules have the same difficulty to cross the
membrane.

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 Polarity and size are the more important molecular features that influence the ability to cross
membranes.
 Small molecules without electric charges, such as CO2, N2, O2, and
 Molecules with high solubility in fat, such as ethanol, can cross membranes almost freely by passive
diffusion (Figure 1).
 Permeability is lower for molecules that have electric charges when the number of positive charges
equals negative charges, such as water and glycerol.
 It might be thought that water can cross membranes freely, but there are some restrictions, and that is
why some membranes contain aquaporins, a type of transmembrane protein with a channel that allow
the water to cross freely.
 The ability of large uncharged molecules, such as glucose, to cross membranes is lower.
 Membranes are highly impermeable to ions and net charged molecules.
 Some values for the permeability coefficient by passive diffusion are: O2: 2.3 cm/s, CO2: 0,35 cm/s,
H2O: 0,0034 cm/s, glycerol: 10 -6 cm/s, sodium and potassium: 10 -14 cm/s.

Permeability

Figure 1. The size and electric features of molecules affect their ability to cross cell membranes (adapted from
Alberts et al., 2002).
 The unequal distribution of ions and molecules between both sides of a membrane leads to the
electrochemical gradients.
 The difference between the inner and the outer concentration of electric charges is known as
membrane potential.
 This gradient is used for many cell functions, such as ATP synthesis and transmission of information
along the nerves.
 Semipermeability is also responsible for osmotic processes, which are water movements across
membranes from a less concentrated solution in one side to a more concentrated solution in the other
side, in order to equal concentrations.
 In this way, plant cells are able to increase in size.
 Molecules that do not cross membranes freely are useful for cells because they can form gradients that
may work as information signals or as tools for other processes.
 Cells have transmembrane proteins that can break gradients by selectively allowing some ions or
molecules to cross the membrane.
 For example, muscle contraction is triggered by the opening of channels that reduce an existing ionic
gradient.
 Transmembrane proteins can also generate gradients by transporting charged molecules and ions
across cell membranes against concentration gradient, avoiding the inner membrane hydrophobic
environment.

 Semipermeability is influenced by the lipid composition.

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 More fluid membranes (see below) are more permeable.

 For example, cholesterol content is important in the plasma membrane.
 An increase in the cholesterol content decreases fluidity and increase hydrophobicity, both features
making the membrane more impermeable.
 Thus, the increase of cholesterol over 30 % (which is a high value) makes myelin membranes very
suitable for insulating the axons and optimizing the propagation of the action potential along the axon.

2. Fluidity
 Fluidity is one of the membrane features.
 It is related to the ability of molecules to move within membranes.
 Higher fluidity means that movements are more frequent.
 Cell membranes are actually a sheet of fat, where molecules are in a semi liquid viscous state.
 Thus, it can be guessed that molecules can move by diffusion.
 A glycerophospholipid located in the external hemilayer of the plasma membrane may have two type of
movements:
i. Lateral, i.e., in the same hemilayer, and
ii. Flip-flop, i.e., jumping from one hemilayer to the other (Figure 2).
 In artificial membranes, both types of movements have been observed, being the lateral movements
much more frequent than flip-flop.
o By lateral diffusion, lipids can travel 30 µm in 20 seconds; they can travel the whole
circumference of a medium size cell in a minute.
o On the other hand, flip-flop movements are really infrequent because the hydrophilic head of
the lipid molecule must cross the internal hydrophobic layer of fatty acid chains, and this is
thermodynamically difficult.
 For one lipid molecule, the probability of a flip-flop shift is about one time per month.

NB: However, cholesterol behaves different and can do flip-flop quite easily.

Figure 2. Membrane fludity allows lipid movement. Lateral movements are frequent, but flip-flop movements
are rare for lipids and have not been observed for proteins.
 Fluidity may change depending on the chemical composition of the membrane.
 Generally, shorter fatty acid and higher amount of unsaturated bonds between carbons of fatty acids
increase membrane fluidity.
 The amount of cholesterol also influences the membrane fluidity, but the net effect depends on
temperature and type of lipids in the membrane.

 Cholesterol has two effects:

i. inhibits the transition to a solid gel state (less fluidity), but also
ii. decreases the flexibility of the unsaturated fatty acid chains.
 In general, it can be said that an increase in cholesterol concentration decreases membrane fluidity,
although at low temperatures the effect is the opposite.

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 Internal membranes of the cell, like those of the endoplasmic reticulum, contain very little cholesterol,
and therefore they are more fluid.
 In addition, cholesterol provides membranes with another property known as hydrophobicity, which
makes membranes more impermeable.

Cell membrane models

Membrane models
 Different molecular composition between the two hemilayers of a membrane, known as membrane
asymmetry, may lead to a distinct fluidity in each hemilayer.
 Depending on the molecular composition, lipids may be in two physical phases:
i. liquid-ordered (less fluidity) and
ii. liquid-disordered (more fluidity).
 The outer hemilayer of the plasma membrane is supposed to be more often in the liquid-ordered phase,
whereas the inner hemilayer is prone to be in the liquid-disordered phase.

 Cells can modify the membrane fluidity by changing the chemical composition.
 For example, bacteria adjust the saturation and length of fatty acid chains so that membrane fluidity
is adapted to the environmental conditions.
 Changes in the concentration of some types of glycerophospholipids, such as
phosphatidylethanolamine, may also modulate fluidity.
 Thus, some insects cannot synthesize sterols, like cholesterol, but they can modulate the fluidity of
their cell membranes by modifying the concentration of phosphatidylethanolamine.

 The inner membrane of span mitochondria needs to work as a strong, impermeable barrier for
generating and maintaining a proton gradient.
 It would be done by an increase in the cholesterol content.
 In this way, the hydrophobicity is higher.
 However, cholesterol decreases fluidity, feature that appears to be very important for the function of the
proteins in this membrane.
 Mitochondria solve the problem with span cardiolipin, an unsaturated phospholipid that increases
hydrophobicity, but does not reduce much the membrane fluidity.

3. Heterogeneity
 Because of fluidity, it can be thought that molecules are randomly distributed and, therefore,
membranes are homogeneous regarding the molecular composition, i.e., they show the same molecular
content and proportion independently of the membrane region.
 This is not true.

 There are restrictions to the lateral diffusion of molecules that causes membrane heterogeneity, which
means that there are regions of a membrane with different molecular composition.
 In non-polarized cells, and at scales larger than 200 nm, plasma membrane looks like homogeneous.
 However, we can find large scale domains, such as synapses of cell junctions.
 At scales below 200 nm, membranes show a patchy organization, so that they are heterogeneous.
 The microdomains of membranes are thought to be around 60 nm to 100 nm in size.
 Lipids and proteins show lateral movements mostly restricted to 60-100 nm areas during a few
milliseconds, and then jump to another adjoining area where they remain for another short time.
 This behavior is called saltatory diffusion.
 It is also known that some molecules practically remain in the same position for long times, whereas
other molecules are free to diffuse laterally.

 Restrictions to lateral movements of molecules may be caused by several mechanisms:

i. interactions with the cytoskeleton or extracellular matrix,

ii. interactions of membrane molecules between each other,

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iii. different densities of local membrane areas (changes in fluidity),

iv. amount of electrical charges, membrane curvature, and different thickness of membrane
domains (Figure 3).

Lateral movement restrictions

Figure 3. Model of the plasma membrane showing the molecular interactions that restrict lateral movements
and generate microdomains.
Inner interactions
 Interactions of membrane molecules between each other constrain lateral movement (Figure 4).
 Both, proteins and lipids movements are influenced by these interactions, leading to the formation of
microdomains with differential molecular composition.
 Depending on molecular proportions, microdomains may have different density:
i. solid,

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ii. liquid ordered and

iii. liquid disordered.
 The most common state is liquid disordered, which is the most fluid.

Figure 4. Interactions between lipids, mostly sphingolipids and cholesterol, may form areas with higher lipid
density. Interactions between lipids and proteins may also generate distinct areas or shells around proteins.
 Sphingolipids and cholesterol may become associated spontaneously between each other, reducing their
motility and increasing the molecular density when compared to neighbor areas.
 A small, distinct group of molecules is formed, like a raft in a sea of lipids.
 Actually, these molecular associations are known as lipid rafts, and are thought to be very abundant in
the plasma membrane, forming a mosaic of microdomains.
 Lipid rafts are very small, between 10 nm and 200 nm, and show a highly dynamic behavior, so that
they can move, grow, shrink, appear and disappear.
 Some experiments suggest that certain types of proteins "feel" more comfortable inside lipid rafts.
 These proteins spend more time inside than outside the lipid rafts, thus they travel for some time
within these microdomains.
 It leads to a segregation of molecules along the membrane, and increases the probability of different
molecules to be close to each other more time than just by chance (diffusion), increasing in this way the
probability of certain molecular reactions.
 It is suggested that a high concentration of certain types of lipids in the lipid rafts forms a distinct
chemical environment that makes easier some chemical reactions or molecular interactions.

 Lipid rafts have been proposed to be only present in the outer hemilayer of the plasma membrane
because it is in this side where sphingolipids are abundant.
 Membrane domains have also been suggested in the membranes of some organelles, and it is thought
that some of their functions rely on these membrane domains.

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 In the inner hemilayer of the plasma membrane, microdomains are formed by electrostatic
interactions between basic cytosolic domains, or divalent cation domains, of proteins and the negative
polar heads of lipids.
 Phosphaotidilinositol is involved in the formation of microdomains associated to proteins.
 By adding or removing phosphate groups in the head of this lipid type, the microdomains can be
modulated.
 Another less known example is the association between phosphatidylinositol bisphosphate and
cholesterol that forms microdomains in the inner hemilayer of the plasma membrane.
 These microdomains facing the cytosol may influence intracellular protein scaffolds.

 Traditionally, it has been thought that there are no interactions between both hemilayers of
membranes, so that they distribute their microdomains independently.
 However, evidences suggest that there are interactions between each other.
 For instance, transmembrane proteins simultaneously affect both hemilayers since they cross the
entire membrane.
 Other way of synchronizing both hemilayers may be mediated by the long fatty acid chains, like some
sphingolipis that can be 24 carbons in length (the normal length is about 18).
 These long chains may be inserted among the fatty acid chains of the lipids of the other hemilayer and
influence the lipid distribution.
 Furthermore, lipid domains with long fatty acid chains are thought to be counterbalanced by short fatty
acid chain lipid microdomains in the other hemilayer, keeping constant the thickness of the membrane.

 Membrane proteins, both integral and associated, may also interact between each other and assemble
into macromolecular scaffolds to facilitate transmission of information, cell-cell recognition, initiate
some enzymatic activities, and cellular movement.
 There are also multimeric proteins, active only when all the subunits are hold together.
 For example, the insulin receptor is made up of four subunits.

Interactions with outer elements

 Integral membrane proteins may also have lateral movements, but they are more restricted than
lipids, mostly due to interactions with the extracellular matrix and cytoskeleton through their
extracellular and intracellular domains, respectively (Figure 5).
 These interactions may keep proteins in small areas of the membrane for a longer time than just by
diffusion.
 Cytoskeleton may also form fences just below the plasma membrane that keep proteins restricted to
small areas.

Movements

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Figure 5. Lateral movements of molecules by diffusion may be restricted by interactions with the
extracellular matrix and cytoskeleton (image on the left), although membrane local densities such as lipid
rafts may affect as well. The organization of the cytoskeleton underneath the membrane restricts the
membrane area where molecules can diffuse (image on the right).
 There is a scaffold of myosin and actin (cytoskeleton proteins) beneath the plasma membrane that
interacts with membrane proteins and restrain their movements.
 Although temporally, these retained proteins may act as obstacles for the diffusion of other
membrane molecules.
 In this way, molecular interactions may lead to immobilize large molecule aggregates or to move them
through the membrane, propelled by the cytoskeleton.
 For instance, cilia contain particular sets of proteins thanks to these type of interactions with the
cytoskeleton.
 When the cytoskeleton is disorganized, the membrane become more homogeneous.

 There are other mechanisms to confine proteins in specific regions of the membrane.
 For instance, the epithelial cells of the digestive tube concentrate some transporters and enzymes in the
apical domain (the free surface), whereas others are confined to the baso-lateral domain.
 It means they are polarized cells.
 The segregation of proteins between the two domains is caused by tight junctions, cell-cell junctions
that form a belt-like structure around the cell that cannot be crossed by proteins.
 This polarization is essential for the intake nutritive substances after digestion.
 The plasma membrane molecules of the outer hemilayer interact with molecules of the extracellular
matrix, such as collagen, proteoglycans, hyaluronic acid, and many others.

 Bending and thicknes

 Cending a membrane is another way to form microdomains.
 The initial bending may be the beginning to generate a vesicle, a cellular extension, the growing of an
organelle, or just to make a fold as a barrier for lateral diffusion.

 Che molecular machinery for bending a membrane has to be recruited to a previous microdomain.
 Some lipids and electric charges concentrated in son membrane sites recruit the bending proteins.
 Phosphoinositides (PI) participate in the this recruitment, particularly PIP2 and PIP3.
 They can change the electric charge of their polar heads quite easily by local chemical modifications.
 Phosphatidilserine is also involved in the beginning of membrane bending when it is transported by
flipases from one hemilayer to the other hemilayer.

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 Both, phosphoinositides and phosphatidylserine can be retained in the bending site by the recruited
proteins.

 Lipid microdomains recruit proteins that effectively bend the membrane (Figure 6).

 They are specialized proteins, such as BAR-domain proteins (Bin/amphyphysin/Rsv161).

 Bending may be induced by two mechanisms:
i. assembling of a scaffold of proteins that pull or push the membrane, or
ii. by inserting amino acid sequences among the lipids as a wedge.
 For example, caveolins cause membrane curvature to form caveloae, tetraspanin force membranes to
form tubules, ESCRT complex helps with vesicle formation within endosomes to form multivesicular
bodies.
 Actin is another strong membrane bending agent by actin filament polymerization that pushes the
plasma membrane outward, leading to cell expansions.
 Many proteins that can curve the plasma membrane also activate actin polymerization.

Membrane bending mechanisms

Figure 6. Membrane bending by molecular mechanisms.

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 There are other microdomains in membranes formed by transmembrane proteins (Figure 7).
 These regions show different membrane thickness because they contain proteins with longer
hydrophobic amino acid sequences that get surrounded by lipids with longer fatty acid chains.
 The proteins and lipids "feel" more comfortable when they are together, since they fit properly their
hydrophobic parts.
 They form membrane domains that exclude other molecules, either proteins or lipids, with shorter
hydrophobic regions.

Figure 7. Longer hydrophobic amino acid sequences of some proteins forms thicker membrane domains.
Bibliography

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