Professional Documents
Culture Documents
P 01
BLOCK -1
CELL & MOLECULAR
BIOLOGY OF PLANTS
UNIT -I
UNIT -II
Editor
Writer
Unit I
1.0
1.1
INTRODUCTION
1.2
OBJECTIVE
1.1.1
1.1.2
1.1.3
1.1.4
CYTOSKELETAL FILAMENTS
1.1.5
LET US SUM UP
1.1.6
1.1.7
ASSIGNMENT/ACTIVITIES
1.1.8
1.1.9
REFERENCE/FURTHER READING
1.2.0
1.2.1
1.2.2
1.2.3
1.2.4
LET US SUM UP
1.2.5
1.2.6
ASSIGNMENT/ACTIVITIES
1.2.7
1.3.0
1.3.1
ENERGY FLOW
1.3.2
LAW OF THERMODYANAMIC
1.3.3
1.3.4
LET US SUM UP
1.3.5
1.3.6
ASSIGNMENT/ACTIVITIES
1.3.7
1.3.8
REFERENCE/FURTHER READING
1.4.0
CELL WALL
3
1.4.1
PRIMARY STRUCTURE
1.4.2
CHEMICAL COMPOSITION.
1.4.3
1.4.4
LET US SUM UP
1.4.5
1.4.6
ASSIGNMENT/ACTIVITIES
1.4.7
1.4.8
REFERENCE/FURTHER READING
1.5.0
1.5.1
1.5.2
1.5.3
FUNCTION
1.5.4
CARRIERS
1.5.5
CHANNELS
1.5.6
1.5.7
RECEPTORS
1.5.8
LET US SUM UP
1.5.9
1.5.10
ASSIGNMENT/ACTIVITIES
1.5.11
1.5.12
REFERENCE/FURTHER READING
1.6.0
PLASMODESMETA
1.6.1
INTERNAL STRUCTURE
1.6.2
FUNCTION OF PLASMODESMETA
1.6.3
1.6.4
LETS SUM UP
1.6.5
1.6.6
ACTIVITES/ASSIGNMENTS
1.6.7
1.6.8
REFRENCE/FURTHER READING
1.1
INTRODUCTION
The cell is the fundamental unit of life, the building block from which all organisms are constructed.
The properties of cell exhibiting the characteristics of life, define both the potential capabilities & the
inherent limitations of all living organisms. The cell theory given in 1839 by German biologist
M.Schleiden & Theodor Schwann was originally based on the observation that different kinds of cells
resemble one another when observed microscopically &functionally. When the properties of many
different cell types was examined , it was found that cells share common characteristic functions carried
out by specialized subcellular structure known as organelles . (figure 1)
The specialized found in plant, shoot, leaves, roots, flower and fruits are classified into three tissue
system ground tissue system ,dermal tissue system and vascular tissue system .Each tissue system
carries different generalize function :the vascular tissue system transport water and solutes to long
distance in the plant, the dermal tissue system provide protection and perform exchange at the surface of
the plant, and the ground tissue system provide cells that carry out photosynthesis , storage and support.
Each tissue system has many specialized cells, a few cell types are found in more than one tissue system.
The different types of specialized plant cell are distinguish by cell shape and by properties of the cell
wall and protoplast. Plant cell wall is one of the most important distinguishing feature of the different
kind of specialized cell. All plant have a thin and flexible primary wall, made of the polysaccharides
cellulose and other carbohydrates . Other cell types have addition to a primary wall, a thick, rigid
secondary wall in which cellulose is impregnated with lignin.
The internal atmosphere of cell differs from that of its external environment. This difference is maintain
throughout the life of the cell by thin surface membrane. The plasma membrane which controls the
entrance & exit of molecules & ions. The capacity of the plasma membrane to act as a selective barrier
between the cell & the medium is called permeability .Most plant cells, have a thick cellular wall that
cover & protects the plasma membrane . Some animal cells are surrounded by a cement like layer called
a cell coat, which generally plays no role in permeability but does have other important functions.
Cell requires chemical building blocks such as sugars, fatty acid, nucleotides & amino acid .Cells need
energy both to drive the chemical reaction involved in building cell components. Most of the chemical
reactions that take place in the cell would normally occur too slowly to maintain life as we know it. Cell
requires a set of information to carry out the reactions.
The extraneous coat of the plasma membrane of the plant cell is known as cell wall. The cell wall is a
rigid and protective layer around the plasma membrane which provides the mechanical support to the
cell. The surfaces of plant and bacterial cells exhibit many of these same properties, but they also exhibit
a few unique features that are not shared by the cells of animals. Plant cell walls provide a supporting
framework for intact plants. In addition to providing mechanical support & strength for the plant as a
whole, the cell wall protects individual cells from osmotic rupture and mechanical injury .The rigid cell
wall also plays a central role in determining the characteristic shapes of plant cells .Although cell walls
were once viewed as relatively inert secretions of the cell they surround, more recent studies have
revealed the wall to be a dynamic structure that carries out many activities. The cell wall also acts as a
permeability barrier and plays a role in certain types of secretary and metabolic events.
Although cell wall provides a thick encasement for the plant cell, this barrier does not seal the cell
completely from its surroundings. Plant cell wall usually contain small opening or plasmodesmeta,
through which adjacent cells maintains direct contact with one another. In electron micrograph
plasmodesmeta appears as a narrow channel in the cell wall that are linned by plasma membrane and
often transversed by a tubular of ER. Thus the plasma membrane, cytosol and ER are all in continuity
from one cell to the next. Plasmodesmeta tends to be concentrated in special areas of the cell wall called
6
as PITS FEILDS, where the primary wall is thinner then the normal and secondary wall is absent.
The main focus of this topic is to learn about the connection between plant cells and how these
connection differs from the those found in the animal cells.
1.2
OBJECTIVE
The main objective of this unit is to understand the basic unit of life and Cell interior
The main focus of this sub topic is to get acquainted with each tissue system and their functions which is
carried out by different type of specialized plant cell.
The main focus of this sub topic is to know about the selective barrier of the cells and their molecular
architecture and the way in which they controls the transport.
The main emphasis of this sub topic is to know about the energy which cell needs to drive the chemical
reaction to maintain life.
The main aim of this sub topic is understand about the primary barrier of the cell along with its
chemical composition and arrangement pattern followed during the growth of the plant.
The main aim to study this topic is to know how cell communicates within plant Are similar structures
are found in animal cells , if yes than how these two can be compared.
1.1.1
Cell maintains a selective barrier called the plasma membrane, a structure measuring 7-8 nm in
thickness that constitute the outer membrane boundary of all living cells .By regulating the passage of
material into & out of cell, the plasma membrane ensures that optimum conditions for living processes
prevail within the cell interior .Plasma membrane also plays an important role in cell -to-cell
communication, transmitting signals from the cell exterior to the cell interior. In plants a rigid cell wall
is found directly outside the plasma membrane .The rigidity and of the cell wall provides the shape of
the cell it encloses and protect the cell from adverse environmental condition . In addition to the
presence of plasma membrane at the outer cell surface, membranes are also employed to partition the
cell interior into multiple compartments which is exclusively found in animal and plant cells. An
elaborated system of interconnected membrane channels and vesicles known as Endoplasmic reticulum
(ER) plays an important role in transporting newly synthesised proteins to various destinations within
the cell. Closely associated with ER is the Golgi complex, a stack of membranes involved in processing
newly synthesised proteins and packaging them into membrane vesicles for storage and secretion.
7
Several other membrane bound organelles also occurs in plants. Lysosomes are small membrane and
enclosed structure that serve a digestive functions to breaking down foreign materials & intracellular
constituents that are no longer needed by the cell. Peroxisomes carry out certain kind of oxidation
reactions and vacuoles, which are especially prominent in plant cells, a large vesicle that functions as
storage compartments & in maintaining water balance. In addition to the preceding membrane -enclosed
organelles, we will see shortly that membranes surround the genetic material and energy transforming
organelles of plant cell
1.1.2
Cells utilize genetic information to guide the synthesis of most of the cell's component. This genetic
information is stored in the nucleus surrounded by a double membrane envelope. The nucleus of plant
cell is largely occupied by a mass of intertwined chromatin fibres, which contains the DNA molecules
which stored most of cell genetic information .A small portion of nuclear DNA is localized in a small
spherical structure known as nucleolus, which contains DNA information involved in the formation of
ribosomes. A fluid like material called nucleoplasma fills the space around the chromatin fibres &
nucleoli condense into compact structure known as chromosomes. The outer boundary of the nucleus is
formed by two concentric membrane that together forms the nuclear envelope. Connections are observed
between the outer membrane of the nuclear envelope & ER. The most distinctive structural feature of
nuclear envelope is the presence of numerous nuclear pores .Nuclear pores help to regulate the flow of
the material between nucleus & cytoplasm. Genetic information and encoded in the cells DNA guides
the synthesis of specific protein molecule. This process of protein occurs on small cytoplasmic granules
known as Ribosomes. Prokaryotic ribosomes are slightly smaller than eukaryotic. Eukaryotic ribosomes
are found free in the cytoplasm and attached to the membrane of ER.
1.1.3
Metabolic reactions occurs virtually everywhere in the cell, but those that are most central to the flow of
energy are localised predominantly in the cytoplasm. Reactions involved in the initial breakdown of
energy rich nutrients occurs in the cytosol, which is the fluid like portion of the cytoplasm that surrounds
the cytoplasmic organelle. Once this initial process is complete, a second set of reactions occurs in
which the most of the energy contained in the nutrients is released and used to drive the formation of
energy rich molecule ATP. Mitochondria are large and enclosed by two membranes. The inner of the
two membranes is folded into a series of CRISTAE that project into the internal cavity, or matrix, of the
mitochondria. They are the site of many chemical reactions, the matrix also contains DNA and
ribosomes. In addition to obtain energy from the breakdown of energy rich nutrients, plant cells are
capable of trapping energy from sunlight & converting into chemical energy. Photosynthesis takes place
in especialized organelle called CHLOROPLAST. Although chloroplast are usually longer than
mitochondria, the structure of the two organelles exhibits some fundamental similarities. Like
mitochondria, chloroplast are enclosed by two membranes surrounding an internal compartment, in this
place designated the stroma. Within stroma are the thylakoid membrane, which contain chlorophyll and
other light absorbing pigments. Like the matrix space of mitochondria, the stroma of chloroplast contain
both DNA and ribosomes.
1.1.4
CYTOSKELETAL FILAMENTS
Plant cells have developed an elaborate network of cytoplasmic filaments, such network of the filaments
are called cytoskeleton. It consist of three distinct components-(i)Microtubules- it is the largest
filaments, which is rigid hollow structures measuring 25nm in diameter and contain tubulin protein.
They are used in the construction of the mitotic spindle that moves chromosomes during cell division.
(ii) Actin filaments or Microfilaments- generates movements within the cytoplasm. They measures 6nm
in diameter and are constructed from actin. Actin filaments generate cytoplasmic streaming.
(iii) Intermediate filament- It is found between that of actin filaments and microtubules. Its protein
composition varies, depending on the cell type. The role of intermediate filaments in motility is yet too
established, but they play important role in structural support and in anchoring.
1.1.5
LETS SUM UP
o All organism are composed of cells that performs the same basic functions.
o All cells are enclosed by plasma membrane, in plant and bacterial cell rigid cell wall is present.
o In Eukaryotic cells intra cellular membrane form a series of cytoplasmic compartments known as
endoplasmic reticulum, Golgi complex, lysosome, peroxisomes and vacuoles.
o In eukaryotic cell genetic information is stored in a membrane enclosed nucleus which is larger and
have a complex internal membrane system.
o In eukaryotic cells mitochondria are involved in brackdown of nutrients and converting their energy
into a useful chemical form.
o The chloroplast of plant cells carry out photosynthesis which traps energy from the sun light.
1.1.6
Organelle
b.
Cytosol
c.
Protoplast
d.
Protosol
b.
c.
d.
3. The elaborate system of interconnecting membrane channels and vesicles are known asa Mitochondria
b . Cell wall
c Plasma membrane
.
d Endoplasmic reticulum
Nucleus
Ribosomes
c.
Plasma membrane
d.
Mitochondria
Vacuoles
b.
Ribosomes
c.
Lysosomes
d.
Protosol
10
1.1.7
ACTIVITIES/ASSIGNMENTS-
Collect different plant material and prepare slides to observe the cell .
1.1.8
1.
2.
3.
4.
5.
1.1.9
FURTHER READINGS
Principles of Cell and Molecular Biology Second Ed. Lewis J.Kleinsmith, Valerie M.Kish.
Parenchyma cell are the generalized multipurpose cell in plant .Most parenchyma cell have thin primary
wall those varies from spherical to barrel like in shape .parenchyma cell store food reserves as a potato
tuber (starch storing parenchyma).the parenchyma cell of green leaves a specialized for photosynthesis
these cell contain numerous large chlorophylls and are called chlorenchyma. Other parenchyma cell are
specialized for the transport of solute through the cell membrane .these cell have the greatly enlarged
area due to the highly convoluted inner surface of the cell. Transfer cell are found in the
ectarines
where the extensive membrane houses transport channel thats secrete sugar and other nectare
component to the exterior of the cell. Collenchyma cell function in the support of the growing tissue.
Individual collenchyma cell are long and narrow and have unevenly thickened primary wall.
Cholenchyma cell for cable of thousand of cell that together can provide membrane support while stem
or leaves elongates. Collenchyma in the veins of the leaves form the strings of celery stages.
Sclerenchyma cell function in the support of tissue that are expanding. Sclerenchyma cell are long
narrow with a thick, hard, rigid secondary wall .unlike parenchyma and collenchyma cells that are living
11
cells sclerenchyma cells are dead at maturity. The cells function in mechanical support is carried out by
the strong cell wall. Sclerenchyma fibres make natural woddy tissue and also form long strands in the
leaves and stem of many plant . Natural fiber ropes such as those from hemp or the sisal plant are made
up of thousands of sclerenchyma cell. Some sclerenchyma cell called sclereids much shorter than fibers,
these form the hard layers of shells and peach pits and small cluster of sclereids form in pear fruits
(Fig I)
PARENCHYMA CELL
COLLENCHYMA CELL
SCLERENCHYMA CELL
12
1.2.2
Epidermal cell form the surface layer of the plant ,The typical epidermal cell are flat and form a
continuous layer with no spaces between the cell .Each epidermal cell secretes hydrophobic polymer
cutin on the surface , which reduce the amount of water lost by evaporation .Most cell also secrete
waxes on the surface of cutin which reduce transpiration as well as wettability of the leaves epidermal
cell of green leaves contain choloroplast allowing light to penetrate for the photosynthesis. Epidermal
cell of petals contain anthocyanin pigment within the vacuoles or oranges carries pigment within the
plastids, giving rise to bright colours. Guard cells are specialized epidermal cell that functions as a small
pores in the plant surface, allowing the carbon dioxide needed for photosynthesis to diffuse from the
external atmosphere into the chlorenchyma tissue .Guard cell are crescent shape , contain green
choloroplast ,and are rapidly change there shape in response to changes in wide status. As guard cells
take up water ,the pore opens and as water evaporates the pore closes. The two guard cells and a pore are
called stomata. Trichomes are hair like cells that project from the surface of plant. They function to
reduce water loss by evaporation and trapping water vapour near the plant surface .In some plant
trichome are glandular and secrete sticky or toxic substance that repel insect herbivores .
(FigII)
STOMATALCELL
13
1.2.3
The vascular tissue system is composed of both xylem and phloem tissue. Xylem functions to carry
water and mineral nutrient absorbed at the root tip throughout the plant, stem and leaves. Vessels
element are the major tissue involved in the transport of water and the solutes. Vessel element are
elongated cells with thick secondary wall and the rigid wall keep the vessel element from collapsing.
Like sclerenchyma vessel element are dead at maturity ,so that each cell are empty tubes .Before vessel
element die the cell protoplast releases enzyme that degrade the cell wall ends of the cell forming a
perforation as a continuous pipe. Other xylem cells called tracheids also function in transport of water
and solute but are efficient because they lack perforations. Xylem tissue also contains sclerenchyma cell
function in support and parenchyma cells that function as transfer cell. Phloem tissue function to
transport the product of photosynthesis from green tissue to parts of the plant that are energy rich
carbohydrates required for storage by a process called translocation. Sieve element are the cell of
phloem that are elongated with primary wall. Phloem sap travel under a positive pressure and the thick
,elastic cell walls allow the cells to adjust fluctuation in pressure over a day night cycle. Sieve element
have large conspicuous pores on the end wall, forming plates. The sieve plate pore allow the phloem sap
14
to travel cell to cell along the file of cell called a sieve tube. Each phloem element is living, with an
intact plasma membrane, and the differential permeability of the membrane prevents solute leaking out
of the sieve tube. Sieve tube element lack some other component of the cytoplasm this feature functions
to keep the pore unplugged. Companion cell parenchyma are associated with sieve elements. The
nucleus of the companion cell must direct the metabolite to the companion cell itself and of its sister
sieve elements.
(Fig III)
15
1.2.4
LET US SUM UP
Each tissue system has many specialized cells which has distinct shapes and cell wall properties.
Parenchyma cells are the generalized, multipurpose cell of the ground system.
In the dermal system Guard cells are the specialized epidermal cells that performs the function of
exchange of gases.
In vascular system xylem and phloem are the specialized cells that carries water and mineral
nutrients absorbed by the roots to the parts of the plants.
16
1.2.5
Epidermal cell.
b.
Trichome.
c.
Vessels.
d.
Phleom.
2. The tissue system which provides protection and exchange at the surface of the plant isa.
b.
c.
Nectaries.
b.
Epidernal cells.
c.
Guard cells.
Sclerenchyma
b.
Parechyma
c.
Collenchyma
Sclerenchyma
b.
Parechyma
c.
Collenchyma
17
1.2.6
ACTIVITES/ASSIGNMENTS-
Prepare charts of each tissue system and descried their cell structure and function.
With the help of mascuration techniques visualize different vessel cells of collected
angiospermic material.
Collect different type of plant material and visualize guard cells and trichome and prepare
record of it.
1.2.7
1.
2.
3.
4.
a.
1.2.8
REFERENCE/FURTHER READING
Burgess , Jeremy, An Introduction to Plant Cell Develop. York: Cambridge University Press.
Gunning , Brian E.S. , and Martin W.Steer. Plant Cell Structure and Function.
ENERGY FLOW
To perform various task cell require energy to grow, move, synthesis & transportation. In scientific
concept energy means capacity to perform work or ability to cause some kind of change to occur. For
cell to form function energy must flow into cell from its surrounding. Energy occurs in different form,
cells obtain energy either in the form of organic molecules that contain chemical energy or energy from
photosynthetic organisms as photons of light. This energy is of two types.
Kinetic energy deals with motion of molecules, thermal radiation etc. Potential energy is the energy
stored in bonds connecting atoms.
18
1.3.2
LAWS OF THERMODYANAMICS
Thermodynamics principle helps us to understand about these energy law & their transformation:
First Law Energy can never be created nor be destroyed however it can be transform from one form
to another .Energy of universe is constant, either change occurs in energy form or it's physical location.
Transformation takes place between various forms of energy such as heat, light, electricity, mechanical
energy & chemical energy. In thermodynamics particular location in which energy changes occurs is
reffered as system & rest of the universe is surrounding. Energy content of an individual system can be
changed, but total energy content of system & surrounding remains constant. In cell 1st law is applicable
in the sense that energy is transformed from surroundings & is transformed into those forms which is
used by cell, for example in autotrophs, light energy is converted into chemical energy which is
essentially used by heterotrophs.
Second Law- 1st law does not tells us about the probability that any such process is actually occuring.
As a biologist one must like to know the direction of reaction & whether energy is released or
absorbed.This law states that energy event in the universe occurs in the direction that cause the system &
surrounding to exhibit a net increase in randomness i.e Entropy. According to second law the Entropy of
system & surrounding always increases but this is not true that entropy of individual system always
increase. It may increase or decrease or remain same .Free energy or G (Gibbs). Describes the
thermodynamic factors that allow us to apply the second law of thermodynamics to a individual
chemical reaction without requiring us to measure the entropy change. It represents the energy that can
be harnessed to do useful work. For living organisms where pressure and volume remains constant, the
change in free energy that accompanies any biological process is determined by two parameters;
(G=negative)
reactions , i.e they realize free energy (products contains less bond energy ) .It indicate the direction of
the reactions i.e breakdown of complex organic molecule into simpler one is exergenic reactions .
If Entropy decreases then free energy increases ;
i.e (S= negative) (G=positive)
(E>0);
(a)
( reactant(eq))
G0'= -2.303 RT log(product(eq)..........
(b)
(reactant(eq)
Equilibrium constant Keq= (product(eq)) ........... (c)
(reactant(eq))
Keq can be substituted in eq (b)
G0'= -2.303 RT log Keq ...........(d)
(Equilibrium constant at pH=7)
If G0'= positive, direction of reaction , reactant -----> product,
reaction is exergenic i.e Keq>1 & (product)> (reactant);
If G0'= positive, direction of reaction , reactant <----- product,
reaction is endergenic i.e Keq<1 & (product)< (reactant);
1.3.3
One widely occurring pattern involves the use of high energy phosphorylated compounds that release
energy when their phosphate groups are removed. Such compounds play a key role in transferring
20
The first law states that the total amount of energy in the universe is constant which means that a
cell`s energy requirements can be met only by taking in energy from the surroundings and
transforming it into useful forms.
The direction in which the chemical reaction proceeds is governed by the second law of
thermodynamics, which states that the events proceeds in the direction that causes the entropy of
the universe to increase. All reactions proceeds in the direction that cause the free energy of the
system to decrease.
ATP plays a central role in coupled reactions , trapping energy released by the oxidation .
1.3.6ACTIVITES/ASSIGNMENTS
Enlist
various
examples
which
fulfills
the
law
of
thermodynamics.
Answer of the above question is already provided in notes. Review your work accordingly.
Principles of Cell and Molecular Biology Second Ed. Lewis J.Kleinsmith, Valerie M.Kish.
21
PRIMARY STRUCTURE
Cell wall is complex in nature .It remains differentiated in the following layers:
(i) Middle Lamella
(ii)Primary cell wall
(iii)Secondary cell wall
(iv)Tertiary cell wall
(i) Middle Lamella-The cells of plant tissue generally remain cemented together by intercellular matrix
known as middle lamella. It is composed of pectin, lignin & some other proteins. They are dissolved in
strong acids.
(ii) Primary Cell Wall- It first formed cell wall. Its thickness is about 100-200nm.It is the outer most
layer of the cell & it forms the only cell wall in the immature meristamatic & parenchymatous cells. It is
comparatively thin & permeable. It is composed of loosely organised network of cellulose microfibrils
associated with hemicellulose pectins & glycoproteins. Pectins are important in imparting flexibility &
makes primary cell wall to expand during cell growth. The cellulose synthesizing enzyme that
synthesize cellulose microfibrils are localized within plasma membrane. These are called rosettes; add
glucose molecules to growing microfibrils. It is composed of polysaccharide cellulose but other
substance may be incorporated in it. Lignin or suberin may also present in it & epidermal cells of leaves
& stem also posses cutin & cuter waxes & are impermeable to water. Primary cell wall of yeast and
fungi is composed of chitn.
(iii) Secondary Cell Wall- It lies after primary cell wall. It is thick, permeable & lies close to tertiary
cell wall or lies close to plasma membrane .It consist of bulk of many layers and at maturity cell wall
consist It gives shape & mechanical strength to the cell .It is chemically composed of compactly
arranged cellulose & lignins.
(iv) Tertiary Cell Wall- It is found beneath the secondary cell wall. It differs form primary and
secondary cell wall, and contains xylans. (FIG I)
22
1.4.2
CHEMICAL NATURE
Plant's cell wall are composed of carbohydrates known as cellulose. Besides cellulose various chemical
substances as hemicelluloses, pectin, lignin, cutin & chitn. The cell wall also contains certain minerals
like calcium & magnesium in the form of carbonates & silicates. The cellulose is a polysaccharide & it
is the most abundantly occurring chemical substance of most plant cell. Chemically cellulose contains
long chains of glucose molecules. Glucose is the structural unit of cellulose & about 3000 glucose
molecule are linked together to form long chain of cellulose molecule. A bundle of 100 chain molecules
of cellulose forces the elementary fibril known as micelle. The 20 micelle when get arranged parallel
form the fibrils of 25nm thick known as microfibrils (5-15nm in diametre).These microfibrils form large
sized bundles 0.5micrometre thick of cellulose fibre to form the macrofibrils. The macrofibrils
consisting of many cellulose fibrils. The hemicellulose is composed of monosaccharide units such as
arabinose, xylose, mannos& galactose. It occurs sometimes in between the macrofibrils of cellulose.
(FIG II)
HEMICELLULOSE
They are heterogeneous groups of carbohydrates polymers constructed from various 5&6 carbons sugars
including xylose, arabinose, mannose & galactose. The hemicellulose xylan which utilizes the pentose
xylose as its main building block, accounts as much as 50% of the cell wall in woody tissues.
Hemicellulose molecule binds to the cellulose microfibrils &to each other creating a coating that helps
to bond these microfibrils together into a rigid interconnected network of cellulose & hemicellulose.
23
(FIG II)
PECTINS
These are the polymers of carbohydrates D-galactouronic acid & D-glucouronic acid units .Like the
glycosoaminoglycane of animal cell, they readily form hydrated gels. This property of pectin molecules is
responsible for gelatin that occurs during the process of making jams and jellies .Pectin molecules are
involved in binding adjacent cell wall together & in forming the matrix in which cellulose microfibrils are
embedded. (FIG II)
LIGNINS
They are group of polymerized aromatic phenols. They occurs mainly in woody tissues, as cross linked
network that contributes to the hardening of the cell wall. Lignin molecules are localized between the
cellulose fibrils, where they function to rest compression forces . (FIG II)
GLYCOPROTEINS
They are important constituent of plant cell wall, accounting as much as 10% of the total mass. Among
the wall glycoprotein a group of common glycoprotein are called extensins. Extensins & other
glycoprotein forms cross linked networks with each other as well as with cellulose microfibrils (FIG II)
OTHER COMPONENT
In addition to the preceding components a small percent of the total mass of the cell wall is accounted
for the lipids, including waxes & other complex polymers .Minerals such as Ca & K also occurs in the
plant cell wall in the form of inorganic salts. (FIG II)
1.4.3
The cell wall originates in the developing cell plate. During telophase of the mitotic cell cycle, the
phragmosome, a flattened membranous vesicle containing cell wall components,froms across the cell
within a cytoskeletal array called the phragmoplast. The noncellulosic cell wall polysaccharides
synthesized in the golgi apparatus and packaged in the vesicles fuse with the growing cell plate. The
plate grows outward until the edges of the membranous vesicle fuse with the plasma membrane, creating
24
two cells. Finally the new cell wall fuses with the existing primary cell wall, The plant golgi apparatus is
a factory for the synthesis, processing and targeting of glycoproteins. Thus except for cellulose the
polysaccharides, the structural proteins, and a broad spectrum of enzymes are coordinately secreted in
golgi derived vesicles and targeted to the cell wall.
Cell enlargement depends on the activities of endoglycosidase, endotransglycosylase or expansin or
some combination of these. But cell shape is largely governed by the pattern of cellulose deposition.
Termination of cell growth is accompanied by cross linking reaction involving proteins and aromatic
substances.
The plant cell surface is covered by a cell wall that provides enormous strength and mechanical
support to the whole plant.
The plant cell is composed of various composition of cellulose, hemicellulose, pectin, lignin,and
glycoproteins.
The cell wall is synthesized in a stepway fashion, sterting with a thin zone called the middle
lamella, follwed by the relatively thin and flexible primary cell wall and finally the much thicker
and the stronger secondary wall.
25
Hemicellulose.
b.
Middle lamella.
c.
d.
2. The molecule that binds to the cellulose creating a coating of rigid interconnecting network isa.
Hemicellulose.
b.
Pectin
c.
Lignin.
d.
Hemicellulose.
b.
Fibrils.
c.
Lignin.
d.
Macrofibrils.
Hemicellulose.
b.
Pectin
c.
Lignin.
d.
Hemicellulose.
b.
Pectin
c.
Lignin.
d.
cellulose.
26
1.4.6 ACTIVITES/ASSIGNMENTS
With the help of staining techniques visualize different cell wall component.
3.
4.
5.
Principles of Cell and Molecular Biology Second Ed. Lewis J.Kleinsmith, Valerie M.Kish.
27
was made by who suggested that membrane was composed of phospholipid of one molecule
thick. It was shown by an experiment in which the phospholipid was spread on water. This
form layer of one molecules thick on water surface. Phospholipid are known as amphiphatic
molecules which contain both hydrophilic & hydrophobic regions .Langmuir interpreted his
model that hydrophilic or head groups of the lipid molecules remain attach to the water
surface and the hydrophobic tails remains free towards the air.(Fig I)
(ii) Lipid Bilayer Model Of Gorter & Grendel- Gorter & Grendel proposed a lipid bilayer model of
membranes structure from the experiments of RBC's. When lipid extracted from RBC's where
spread on the water surface, it was found that lipids were also spread as one layer on water. But it
covers twice the area on the water surface than that the area of the cell from which lipid is extracted.
The model of Gorter & Grendel gives a new impetus to membrane research as they first tried to
describe the structure of membrane at the molecular level. .(Fig II)
(iii) The Danielli-Davson Model-They concluded that biological membrane could not be of lipid alone.
Danielli & Davson proposed a molecular model of the membrane in which hydrophilic had groups
of the lipid molecules is covered on both of the side by a protein layer .The proteins are attached to
the hydrophilic head groups by lipid bilayer by ionic bonds . But in this model , the distance between
ends of the fatty acid chains (hydrophobic tails) is not specified . .(Fig III)
28
(iv) Robertson's Model Or Unit Membrane Hypothesis:-The presence of common structure in all
biological membrane led to postulate Unit membrane hypothesis .For detailed study of the
membrane structure & its molecular organisation , Robertson selected mylein as its experimental
sample .He selcted mylein rather than typical membrane because in case of mylein , multiple layers
of membrane are present which forms quasi-crystalline structure .He carried out investigations on
electron microscope using different stains for lipid & proteins . He found that both lipid & proteins
are present in the membrane .Lipid are present in two layers covered by proteins with lipid head
groups projecting outward towards both membrane surfaces .Robertson's observation corroborates
the structure proposed by Danielli & Davson .The electron microscopic observations & X-rays
diffraction data confirmed the Danielli & Davson model of membrane structure . .(Fig IV)
(v) Fluid Mosaic Model- In this model the main component is the lipid bilayer with hydrophilic groups
oriented towards outside & the hydrophilic groups towards inside of the layer . The basic
requirement for the basic requirements of the molecular organisation of the membrane is free energy.
The term fluid is given because the lipid layer is present in the fluid state. The transition of fluid
layer from non fluid (gel) conditions to a liquid crystalline (fluid) state depends on the temperature
of the cell. According to this model, proposed by SJ Singer & Jhon Nicholson, the principle of
membrane organisation is as follows:
1. Lipids are present in two layers.
2. Proteins are arranged in two ways:
a) Some are embedded in lipid layer, called integral proteins &
b) Some are present on surface of the lipid bilayer, called the peripheral proteins.
3. The lipid layer is usually in liquid crystal line, i.e., fluid state. .(Fig V)
29
1.5.3 FUNCTION
It is selective permeable in nature which allows only selectable molecules to pass through it.
Substance that pass through it by simple diffusion,facilitated diffusion and by active transport
method.
Simple diffusion deives the net movement of dissolved solutes as well as water molecules and
the process is termed as osmosis.
Faciliated diffusion refers to the assisted movement of a substance down its electrochemical
gradient.
1.5.4 CARRIERS
Carriers do not catalyze ATP-hydrolysis. In other words, the transport process do not involve
chemical modification of any of the compound bound to the carrier. Rather carrier catalyze the
movement of inorganic ions and simple organic solutes across the membrane. One defining feature
of the carrier is that they display saturability when the kinetics of transport are expressed to the
substrate concentration. The array of ions and solutes translocated by carrier is vast. The principal
inorganic nutrients including NH4+/ NO3-,Pi,K+ and SO4- are all transported into the cell by
plasma membrane carrier. Carriers are also responsible for taking up ions that play less roles in
metabolism like Cl- ions. The organic solutes translocated into the cell by carrier
are the
fundamental building blocks of polymers: sugars, amino acid, puriens and pyrimidines. .(Fig I).
Active transport involves membrane carrier that moves substance against an electrochemical
gradienti.e. an energy requiring process.The energy releasing activity includes the hydrolysis of high
energy molecules such as ATP, the absorption of photons of light, the transfer of elrctrons and the
movement of other substance down their respective electrochemical gradients.The active transport of
sodium and potassium ions is the classic example of an energy requiring transport system powered
by the hydrolysis of ATP. In most cells the concentration of potassium is higher than the
concentration of the sodium.Maintaining this unequal distribution of sodium and potassium is
important because potassium is needed inside the cell for the activity of certain enzymes and the
30
sodium is required for the active transport of sugar and amino acids.
1.5.5 CHANNELS-
Channels mediates the facilitated diffusion of small ions. As diffusion through ion channels does not
require complex conformational changes it is significantly faster than transport mediated by protein
carrier. Channels are simply open holes in the membrane. The two important points that distinguish
these channels from other openings are- they are capable of discriminating between different ions;
thus separate channels exist for transporting important cellular ions such as sodium. Calcium
potassium and chloride. In addition channels have gates that can be regulated in response to
appropriate stimuli.The ability of channels to regulate the diffusion of specific ions across cellular
membrane plays a significant role in many types of cellular communication.The outward rectifier
KCO1 is a member of the two-pore K+ channel family and is sensitive to the concentrations
of cytosolic Ca2+. An outward rectifier, KCO1 has been identified at the molecular level from an
Arabidopsis EST, based on the highly conserved P-domain motif TxGYGD. This channel is a
member of the two-pore K+ channel family, so-called because there are two P-domains in each
subunit. In contrast to Shaker-type channels, KCO1 has only four transmembrane spans in each
subunit. Two Ca2+-binding motifs called EF hands reside toward the C terminus of the protein. The
functional attributes of KCO1 as an outward rectifier have been confirmed by heterologous
expression. Analysis of these currents and the underlying channels demonstrates that although these
are cation-selective, they do not select as effectively for K+ as the inward rectifiers do. Such
channels probably constitute a major pathway for Na+ uptake into plant cells and could have
important implications for salinity tolerance. These channels are partially blocked by external Ca 2+.
An even less selective channel has been observed in the plasma membrane of xylem parenchyma
cells. This channel is almost as permeable to anion as to cations; such low selectivity is unusual in
plasma membrane ion channels. Perhaps this channel could provide a pathway for release of salts
from the symplasm into the xylem.
Monovalent cation channels at the vacuolar membrane are Ca2+-sensitive and mediate
vacuolar K+ mobilization. Both types ofchannels activate instantaneously in response to an
31
imposed voltage. The fast vacuolar (FV) channel, which exhibits little selectivity among monovalent
cations, in inhibited when cytosolic Ca2+ concentrations exceed 1 pM and is activated when
cytosolic pH increases. Conversely the vacuolar K+ (VK) channels, which is highly selective for K+
over other monovalent cations, is activated by cytosolic Ca2+ in the nanomolar to low micromolar
concentration range and is inhibited by increasing cytosolic pH. A wide range of metabolic and
developmental signals in plants trigger an increase in cytosolic free Ca2+. Thus, the increase in the
concentration of free Ca2+ activates downstream targets that further transducer the initial signal into
the end response. Voltage-gated Ca2+-permeable channels reside in the plasma membranes of a range
of cell types and have been characterized by both patch clamp and planar lipid approaches. The
channels exhibit various degrees of selectivity for Ca2+ over K+-from about 2:1 to 20:1. The channels
are activated by membrane depolarization.
Channels can interact in two fundamentally different ways, First passage of an ion (usually Ca 2+)
through one channel can result in concentration changes that gate a different channel. Second,
channel opening can change Vm, leading to activation of voltage-gated channels. Both aspects of
channel interaction are combined in membrane-based signaling pathways that transduce and amplify
incoming signals. .(Fig I)
Plant cell inward rectifiers are members of the Shaker family of voltage-gated channels. Plant
inward rectifier subunits are products of a multigene family, members of which exhibit tissuespecific expression. One member, KAT1 is expressed selectivity in guard cells another, AKT1 is
found in roots and hyadothodes. The AKT1 gene of Arabidopsis has been disrupted by using T-DNA
insertional mutagenesis. The resulting mutant, referred to as AKT1-1, has dramatically reduced K+
uptake less growth in media with low K+ concentrations. The analysis of K+ uptake in ATK1-1
mutants indicates that AKT1 encodes a high-affinity K+ channel.
Several structural features define plant inward rectifying channels as embers of the Shaker family, a
32
super family
33
1.5.7RECEPTORS
34
Intracellular receptors can act as ion channels. Other receptors are located in intracellular
membranes and can act as Ca2+ channels. The most well-known receptor in this class binds the
second messenger inositor 1, 4, 5-triphosphate. Channels for another second messenger, cyclic ADPribose (cADPR), have been reported recently. Occupation of the receptor (which may be composed
of four subunits) leads to the opening of Ca2+ channels and an influx of Ca2+ intot he cytoplasm from
the vacuole and the ER, each of which contains Ca2+ many orders of magnitude greater than the
cytosolic concentrations. In contrast to plasma membrane-bound receptors, these protein subunits
each have four membrane-spanning domains. Other membrane-spanning proteins also may have
important functions in signal transduction.
Not all tissues or cell types are able to respond to all signals. For example, fruit tissues become
sensitive to ethylene at a certain stage of ripening, whereas guard cells are totally insensitive to high
concentrations of the gaseous hormone. Different responses by different tissues to the same signal
can in part be explained by families of receptors. Auxin, for example can induce perieyel cells to
form adventitious or lateral rots, but in coleoptile cells it promotes elongation. Different receptors
are probably involved in each response. However, divergent downstream elements of the signal
transduction pathway may also distinguish the developmental responses to auxin exhibited by
different cell types. Tissue-specific signals transduction pathways are thus defined not only by the
presence or absence of receptors but also by the presence or absence of downstream apparatus
required to transduce the responses. Tissues can adapt or desensitize themselves to continuous
signals, and receptor concentrations can change during development. For example, when etiolated
seedlings are exposed to red or white light, the cellular concentrations of phytochromes decrease
rapidly.
The gas ethylene regulates ripening, germination, elongation, senescence, and pathogen responses.
Several ethylene receptors have been cloned through isolation of ethylene insensitive mutants and
subsequent use of molecular technology to identify the mutant gene. ETR1, a 79-kDa protein with a
transmembrane domain, was the first receptor cloned from Arabidopsis. The C terminus of ETR1 is
homologous to a bacterial two component system hybrid kinase. ETR1 exists as a dimer in the
plasma membrane. Ethylene joins the two monomers together and permits intermolecular
phosphorylation. Mutations in ETR1 (designated efr1) lead to loss of physiological sensitivity to
ethylene. ETRI has also been expressed in yeast to demonstrate that the protein binds ethylene with
35
high affinity. Competitive ethylene antagonists inhibit this binding. Expression of efr1 in yeast leads
to loss of ethylene binding, confirming that ETR1 is thus a true ethylene receptor. Genes encoding
other ethylene receptors have also been identified, including ERS (ethylene response sensor), Nr
(never ripe, .1 developmentally regulated gene from toma to; and LeTAE1 (a tomato ETR1 homolog
expressed during flower and fruit senescence.)
o Many auxin-binding proteins have been detected, but whether they represent receptors for
different auxin-mediated processes is still uncertain. Indole 3-acetic acid (IAA, reterred to here as
auxin) is a growth regulator with a wide variety of functions in cell division and expansion. Auxin
has been studied intensively for the past 50 years and, not surprisingly, receptors for the auxin signal
have been actively sought. Conventioanl pharmacological techniques have uncovered one wellcharacterized auxin-binding protein (ABP1). The possible receptor function of this protein was
controversial for many years but has recently been established.
Phytochorome, a clearly identified receptor for red light, has protein kinase activity in
cyanobacteria. Phytochromes form a family of 120-kDa proteins. The photoreactive moiety
(chromophore) of these proteins is an open-chain tetrapyrrole. Two forms of phytochrome, A and B,
can each form dimers in solution and physiological evidence suggests that both may dimerize in
vivo.
.(Fig I)
Membrane phospholipids forms a bilayer in which their hydrophobic tails are buried in the
membrane interior and their hydrophilic head groups are exposed at the membrane surface.
Substance that pass through it by simple diffusion,facilitated diffusion and by active transport
method.
36
Simple diffusion drives the net movement of dissolved solutes as well as water molecules and
the process is termed as osmosis.
Faciliated diffusion refers to the assisted movement of a substance down its electrochemical
gradient.
Active transport is carried out by membrane transport proteins that are capable of moving
substance against the electrochemical gradient. The energy required for active transport can be
provided by the hydrolysis of energy rich compounds.
Membrane transport proteins that carries out facilitated diffusion are divided into carrier and
channel proteins.
Channel proteins forms small channels through which specific ions can pass.
37
1. The unit membrane model was proposed bya Daniel and Davson.
b. Robertson
c Singer and Nicholson
d. All of the above
2. Lipid bilayer concept was propounded bya
b. Robertson
c Singer and Nicholson
d. All of the above.
3. The main function of the plasma membrane is a
Protection.
b. Active transport
c Passive transport
d. All of the above.
4. Faciliated diffusion of small ions is carried out bya
Pumps
b. Receptors
c Carriers
d. Channels.
5. RLKs are the example of thea
Pumps
b. Receptors
c Carriers
d. Channels.
1.5.10ACTIVITIES/ASSIGNMENTS-
38
Design small experiment like patoto osmoscope to explain the function of plasma membrane
2.
3.
4.
5.
Principles of Cell and Molecular Biology Second Ed. Lewis J.Kleinsmith, Valerie M.Kish.
Gennis, R.D . Biomembranes: Molecular Structure and Function, Springer-Verlag New York.
1.6.0 PLASMODESMETA
In vascular plants the basic plasmodesmal structure is a tube of plasma membrane surrounding a strand
of modified ER, with particulate material between them. Fig I interprets the ultra structure of vascular
plants in a cross and longitudinal sections. Essental features of plasmodesmeta is a cell to cell tubule of
the plasma membrane that surrounds a cylindrical strand of tightly furled ER, the desmotubule. A thin
darkly stained central rod occupies the center of the desmotubule. A cytoplasmic sleeve or the
cytoplasmic annulus lies between the desmotubule and plasma membrane is considered as possible
pathway for the cell-to-cell water and solute movement. The desmotubule is essentially a solid strand of
lipid, the central rod is composed of the lipid polar groups and a few proteins that can physically occupy
the inner core of the tightly furled lipid bilayer. Much of the cytoplasmic annulus is occupied by
proteinaceous material. These particles are associated with both the outer surface of the desmotubules
39
and the inner surface of plasma membrane. In cross section, 7-to-9 gaps occurs between the particles.
The distance across the gaps, is about 2-3nm as comparable with the 4nm channel diameter. These gaps
are the physical basis of cell-to-cell transport. A wide range of plasmodesmal morphologies have been
observed, within the same plants. These include differences in length, branching and size of the central
cavity. The functional significance of the variations is largely unknown.
(FIG I)
1.6.2
FUNCTIONS
It plays important role in cell to cell communication. To investigate this function microelectrodes have
been employed to monitored the flow of electric current from cell to cell in plant tissue (as in case of
animals gap junctions.). Such studies reveals that electric current passes between plant cells linked by
plasmodesmeta more readily than it does between the cells that are not linked by plasmodesmeta. The
magnitude of the current flow is directly related to the number of plasmodesmeta present. This suggest
that it plays role in cell-to-cell communication, comparable to that played by gap junctions in animals.
40
1.6.3
In plants junctions are termed as plasmodesmeta, whereas in animals it is termed as cell junctions.
In plants plasmodesmeta's are found in pit-feilds where primary cell wall is thin and secondary cell
wall is absent. In animals 3-types of junctions are found(i) Tight junction- which creates the permeability barriers across the layers of cell.
(ii) Plaque bearing junctions- which stablizes the cells against the mechanial stress.
(iii) Gap junctions- which prevents the movement across the cells.
Gap junctions in electron micrograph appears as regions in which the plasma membrane of the two
adjacent cells are aligned in parallel and separated by a tiny gap of the 3nm.Plasmodesmeta electron
micrograph reveals that these are narrow channels in the cell wall that are linend by plasma
membrane and transverse by the tubules of ER.
Connexin proteins are observed on the membrane surface o the gap junctions, which are clusters of 6
connexin molecules surounded by aquoues channels. No such structures are seen in plasmodesmeta.
Permeability of the gap junctions can be altered (closed and opened) by the changes in the
membrane potential, intracellular pH and cyclic AMP levels. Plasmodesmeta permeability is
subjected to regulation by the internal conc. of Ca ions, but its mechanism is not well understood.
In addition to their role in electrical coupling, gap junctions may have developmental and metabolic
functions. No such functions are performed by plasmodesmeta.
Channels in the cell wall known as plasmodesmeta permits the direct cytoplasmic connection for
communication between the adjacent cell.
They are present on the specialized areas of the cell wall called pit fields
In the area around the pit field primary wall is thinner and secondary wall is absent.
Unlike plasmodesmeta in animal cells gap junction permits small ions and molecules to pass
directly between adjacent cells.
42
Answer of the above question is already provided in notes. Review your work accordingly.
Principles of Cell and Molecular Biology Second Ed. Lewis J.Kleinsmith, Valerie M.Kish.
43
UNIT-II
CHLOROPLAST
2.1
INTRODUCTION
2.2
OBJECTIVES
2.1.1
2.1.2
GENOME ORGANIZATION-
2.1.3
2.1.4
2.1.5
RNA-EDITING
2.1.6
NUCLEOCHLOROPLASTIC INTERACTION.
2.1.7
LET`S SUM UP
2.1.8
2.1.9
ACTIVITIES/ASSIGNMENTS.
MITOCHONDRIA
2.2.1
STRUCTURE ORGANIZATION:
2.2.1.1 ISOLATION FRACTION
2.2.1.2 FUNCTIONS
2.2.2
2.2.3
BIOGENESIS.
44
2.3.0 VACUOLES
2.3.1
TONOPLAST MEMBRANE
2.3.2
FUNCTIONS2.3.2.1 DIGESTION
2.3.2.2 pH AND IONIC HOMEOSTASIS
2.3.2.3 DEFENCE AGAINST MICROBIAL PATHOGENS AND HERBIVORES
2.3.2.4 SEQUESTRATION OF TOXIC COMPOUNDS
2.3.2.5 PIGMENTATION.
2.3.3
LET`S SUM UP
2.3.4
2.3.5
ACTIVITIES/ASSGINMENTS
2.3.6
2.3.7
2.1
INTRODUCTION
A Leeuwenhoek observed the presence of chloroplasts in algae and, by the year 1800, chloroplasts had
been identified in the leaves of a wide variety of plant cells. Shortly thereafter it was discovered that
chloroplasts rupture when placed in a hypotonic solution, suggesting that they are bounded by a
semipermeable membrane.
The first insight into the biological function of chloroplasts was provided in 1894 by the German plant
biologist, Thomas Engelmann.In the algae Spirogyra chloroplast winds from one end of the cell to the
other. When the algal cells were illuminated with a finely focused beam of light and examined by light
microscopy. the oxygen-seeking bacteria were seen to congregate near those regions of the cell where
the chloroplast had been illuminated. It was therefore concluded that the light-absorbing and oxygenproducing steps of photosynthesis are housed within the chloroplast.
Mitochondria means Thread like granules found in all types of cells except prokaryotic cells and RBC
of mammals. first systematic studies was initiated in 1850 by Rudolph Klliker who studied them in
cytoplasm of muscle cells. He isolated these particles from shredded muscle tissue and discovered that
they swell when placed in heater leading to conclusion that these particles are surrounded by
semipermeable membrane.
mitochondria, but as cell consume oxygen the colour gradually disappears. Such changes indicate a
45
2.2 OBJECTIVESTo get acquainted with the double membrane organelle performing the function of photosynthesis
To understand the characteristic feature of the eukaryotic organelle frequently termed as
POWERHOUSE of the cell
To learn and understand the plant vacuoles as multifunctional compartments.
2.1.1 STRUCTURAL ORGANIZATIONWhen viewed with the light microscope, chloroplasts typically appear as oval or spherical structures
exhibiting a prominent green pigmentation. They occur in the leaves and other green tissues of higher
plants, as well as in algae. The chloroplasts of leaf cells typically measure about 2-4 11min diameter and
up to 10 11min length, which is roughly twice the size of a typical mitochondrial profile. The
chloroplasts of algae may even be larger, and often assume unusual shapes such as spirals, cups, and
circular bands that wind around the cell. In. algae as few as one or two large chloroplasts are present per
cell, but plant cells typically contain several dozen or more. These multiple chloroplasts appear to be
largely independent of each other rather than forming an interconnected network, as often occurs in
mitochondria. Relatively little internal structure can be seen when chloroplasts are viewed with the light
microscope, although early light microscope did detect tiny green granules within the chloroplast, which
they named grana. Electron micrographs revealed that the organelle is surrounded by a membrane
envelope. This chloroplast envelope consists of two closely apposed membranes, termed the outer and
inner membranes, which are separated from each other by an intermembrane space. The chloroplast
envelope encloses the interior space, or stroma, of the chloroplast, which is analogous to the
46
mitochondrial matrix. Within the stroma is another set of membranes, called thylakoid membranes,
which are organized as a system of flattened sacs. The internal space bounded by the thylakoid
membranes is called the thylakoid lumen. Thylakoid membranes are organized in two different ways,
referred to as the stacked and unstacked configurations. In stacked or grana thylakoids, thylakoid sacs
are stacked upon each other like a pile of coins, generating large membrane masses that correspond to
the grana observed by light microscopy. Grana stacks are connected to one another by membrane-bound
channels referred to as unstacked thylakoids. Each unstacked thylakoid is a large flattened sheet that
makes connections to many or all of the individual thylakoids of a given granum stack. In this way, the
individual thylakoid lumens of 0each granum stack become interconnected 'both with one another and
with the thylakoid lumcns of other granum stacks. Hence the entire thylakoid membrane system of the
chloroplast defines a single, enormously complex, membrane enclosed compartment.
Negative staining has revealed that the outer surfaces of thylakoid membranes are studded with
spherical CFt particles, which resemble the Fl particles observed on mitochondria envelope, while
mitochondrial cristae are continuous with the inner membrane surrounding mitochondrion. As a result,
chloroplasts consist of three separate compartments (intermembrane space, stroma, and thylakoid
lumen), whereas mitochondria have only two compartments (intermembrane space and matrix).(Fig I)
47
(Fig II)
48
2.1.1.2 FUNCTIONS
The most important function of chloroplasts is to carry out the process of photosynthesis. The
process includes the light reaction and dark reactions.
The light reaction takes place in the grana of the thylakoid membrane. In light reaction NADPH
and ATP are produced.
In this reaction reducing power of NADPH is used to fix co2 into carbohydrates.
49
50
with
Thus the small subunit in the chloroplast. Verification of this conclusion has come from experiments
showing that isolated chloroplasts synthesize the large but not the small rubisco subunit. Moreover
the gene coding for the large rubisco subunit has been identified in chloroplast DNA, thereby
providing further support for the idea that this polypeptide is synthesized in chloroplasts. although
rubisco is a soluble protein located in the chloroplast stroma, many polypeptides synthesized by
chloroplasts have turned out to be thylakoid membrane proteins. Among them are subunits of protein
complexes involved in photosynthetic electron transfer (photosystem II, the cytochrome b 0f complex
and photosystem I) and subunits of the ATP-synthesizing CF0-CF1 complex.
Chloroplasts also synthesize some of the
polypeptides found in chloroplast ribosomes and chloroplast RNA polymerase. As in the case of
mitochondria, all the polypeptides synthesized by chloroplasts are encoded by chloroplast DNA.(Fig
III)
52
the inner membrane into the stroma. The tubules then spread into flat sheets that line up in parallel to
form thylakoids. At the same time, chlorophyll and other components of the photosynthetic electron
transfer chain are produced and incorporated into the thylakoids. If developing plant seedlings are
allowed to grow in the dark, a different sequence of events takes place. The proplastids still enlarge and
membrane sheets invaginate from the inner membrane, but these sheets condense into a set of
interconnected membrane tubules known as a prolamellar body. The membranes of a typical prolamellar
body arc arranged as highly ordered hexagonal arrays of membrane tubules. At this stage the plastid is
called an etioplast. l.ksides developing from proplastids in dark-grown seedlings, etioplasts also arise
when green plants containing mature chloroplasts are transferred from light to darkness. Under such
conditions, the thylakoid membranes break up into tubules that fuse together to produce a typical
prolamellar body converting the chloroplast into an etioplast. They act as chloroplast precursors that can
develop into functional chloroplasts when ex- posed to light. Within a few hours after an etioplast is
illuminated, the prolamellar body becomes transformed into an irregular mass of tubules; membrane
sheets then grow out from the tubules and develop into thylakoid membranes. This conversion of an
etioplast into a chloroplast is accompanied by the formation of components of the photosynthetic
electron
transfer
chain,
such
as
chlorophyll.
Etioplasts
contain
high
concentrations
of
protochlorophyllide, which is a metabolic precursor of chlorophyll. When etioplasts are illuminated, the
absorption of light by protochlorophyllide triggers its rapid conversion to chlorophyll.
translation to stop, resulting in the truncated protein. The deamination is carried out by an l{NAbinding enzyme which, in conjunction with a set of auxiliary protein factors, binds to a sequence
immediately downstream of the modification position within the mRNA (Smith and Sowden, 1996).
Although not common, RNA editing occurs in a number of different organisms and includes a
variety of different nucleotide changes. Some editing events have a significant impact on the
organism: in humans, editing is partly responsible for the generation of antibody diversity
(Neuberger and Scott, 2000; Section 12.2.1) and has also been implicated in control of the HIV-1
infection cycle (Bourara et aI" 2000). One particularly interesting type of editing is the deamination
of adenosine to inosine, which is carried out by enzymes called adenosine deaminases acting on
RNA (AGARs) (Reenan, 2001). Some of the target mRNAs for these enzymes are selectively edited
at a limited number of positions, These positions are apparently specified by double-stranded
segments of the pre-mRNA, formed by base-pairing between the modification site and sequences
from adjacent introns. This type of editing occurs for example, during processing of the mRNAs for
mammalian glutamate receptors (Scott, 1997). Selective editing contrasts with the second type of
modification carried out by ADARs, in which the target molecules become extensively deaminated,
over 50% of the adenosines in the RNA becoming converted to inosines. Hyperediting has so far
been observed mainly, but not exclusively, with viral RNAs and is thought to occur by chance, these
RNAs adopting base-paired structures that fortuitously act ns substrates for ADAR. It may, however
physiological importance in the etiology of disease caused by the edited viruses. This possibility is
raised by the discovery that viral RNAs associated with persistent measles infections (as opposed to
the more usual transient version of the disease) are hyperedited (Bass, 1997)
Nuclear
Genes
Cooperate
with
Chloroplast Genes in Making Chloroplast proteins chloroplasts carry less then 10 percent of the
genetic information required to assemble chloroplast. Most of the polypeptides needed by
chloroplasts are encoded by nuclear genes, synthesized on free cytoplasmic ribosomes, released into
the cytosol, and imported into chloroplasts using the targeting mechanisms. Polypeptides destined
for uptake by chloroplasts contain signal sequences that bind to receptor proteins located in the outer
membrane of chloroplast. After binding, the polypeptide chain is inserted into or transferred across
54
polypeptides occur in the nucleus while others reside within the chloroplast the question arises as to
how the expression of the genes located in the organelles is coordinated with those residing in the
nucleus. This issue is especially important for proteins that require polypeptides encoded by genes
located in two different compartments for example rubisco requires one polypeptide encoded by
chloroplast DNA and one polypeptide encoded by nuclear DNA. The mechanism that coordinates
the synthesis of polypeptides made in separate locations but destined for assembly into the same
protein is not well understood. In chloroplasts, some insights have emerged from studies involving
the synthesis of the large and small subunits of rubisco. If an inhibitor of cytoplasmic protein
synthesis is added to intact cells to block formation of the small rubisco subunit by cytoplasmic
ribosomes, synthesis of the large rubisco subunit by chloroplasts eventually slows down as well,
even though the inhibitor has no direct effect on protein synthesized within the organelles are
manufactured in the cytoplasm and then imported? The answer to this question may lie in the
evolutionary history of chloroplasts.
in some instances the nuclear sequences are functional and in other cases the chloroplast sequences
are functional.
Inside the chloroplast contains a mixture of stacked and unstacked thylakoid membrane that
enclose a compartments known as the thylakoid lumen.
The compartment between the thylakoid and the chloroplast envelope is the stroma.
The light reactions of photosynthesis occur in the thylakoid membrane while the dark
reaction occurs in the stroma.
Chloroplast contains their DNA molecules which are transcribed into ribosomal, transfer and
messenger RNA`s.
Chloroplast DNA`S codes for120 or more products including 4rRNA`S 30 or more tRNA`s
20 ribosomal proteins, 4 subunit of chloroplast RNA polymerase, the largest subunit of rubisco, and
several dozens polypeptide components of the cytochrome b6f complex , photosystem I and II and
chloroplast ATP synthase.
Genetic information encoded in chloroplast
DNA`S is transcribed and translated using RNA`s polymerase , ribosomes , and tRNA`s that are
different from those used in the
transcripition and translation of information
Most of the
encoded by nuclear genes , realeased into cytosol, and imported into chloroplast.
56
57
1.
58
2.1.9 ACTIVITES/ASSIGNMENTS
2.1.11 REFRENCES59
BUCHANAN. GRUISSEM.
JONNES.
2.2 MITOCHONDRIA
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2.2.1.1
The first successful technique for separating inner and outer membrane developed by Donald
Parson. In this procedure mitochondria are placed in a hypotonic solutions exposed to the detergent
digitonion until the outer membrane ruptures, releasing the contents of the inter membrane space into
solution. The inner membrane, outer membranes and components of inter membrane space are then
separated by centrifugation. Isolated inner and outer membrane can be readily distinguished from each
other by electron microscopy. Outer membrane book like empty sacs and inner membranes from
vesicles called mitoplast containing matrix material within. Mitoplast can be further fractioned into inner
membrane and matrix component by treating with detergent Lubrol. Lubrol distrups inner membrane
into small inner membrane vesicles containing F1 particles. Biochemical studies of isolated fraction.
provides important information regarding the location of various metabolic activities of
the
mitochondria.
2.2.1.2
Function
Outer membrane of Mitochondria is permeable and contains enzymes involved in lipid metabolism.
It contains monoamine oxidase enzyme Fatty acyl Co-A synthetase glycerolphosphatase acyl
transfrase & Porin.
Inner Mitochondrial is the main site of Mitochondrial ATP formation and it is semi permeable in
matrix .
Inner Mitochondrial membrane is also the site of several enzymatic pathways that are not directly
related to energy metabolism e.g. involves in the synthesis of steroid harmones.
2.2.2..GENOME ORGANIZATION
The genetic evidences indicate that mitochondria contain genes distinct from those located in the
nucleus. Mitochondrial DNAs was identified on the basis of its unique density. The amount of
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mitochondrial DNA found in eukaryotic cells varies. In animal mitochondrial DNA-typically accounts
for 0.1 to 1.0 percent of a cells total DNA contents in Yeast its value is high as 15%. The DNA located
in mitochondria exhibits several features that distinguished it from nuclear DNA. One major difference
is that mt DNA lacks histone and hence not packaged into nucleosomes.
Mt DNA is typically Circular. The mitochondrial DNA circles of animals cells are the smallest about
15,000-20,000 bp in size. Mt DNA of fungi and protists are larger i.e. 20,000-1,00,000bp and plant mt
DNA are largest ranging from 2,00,000bp to million bp in length. One to the tendency of large circular
DNAs to break during isolation many reports suggest that plant mt DNA is linear rather than circular.
Few unicellular prokaryotes such as Paramecium have linear mol of mt DNAs.
In animals, fungi and protists mitochondrial contains few dozens to several 100s mol of DNA, all of
which are identical.
The first direct evidence for an association between cytoplasmic inherited tracts and mt DNA was
produced by Piotr Solnimski et.al. they demonstrated that mt DNA obtained from yeast bearing petite
mutation has a different density than the mt DNA of normal yeast Petites is induced by treatment with
ethidium bromide as a model system. These studies revealed that ethidium bromide stimulates the
breakdown of mt DNA and inhibits mt DNA replication. Long term exposure to ethidium bromide
causes the complete destruction of mt DNA, but if treatment is stopped before all the DNA has been
degraded, the remaining DNA resumes replication until the total context of mt DNA reaches normal
levels. But the new mt DNA is grossly abnormal containing few of its sequence present in the original
mt DNA. The presence of abnormal mt DNA in petite yeast
responsible for the abnormalities in mitochondrial structure and respiratory activity observed in petites
mutants.
2.2.2.1
Mitochondrial DNA-replication
The replication of mt DNA is carried out by DNA p-ax that operates independently of nuclear DNA pase, as mt DNA synthesis can take place anytime during the cell cycle i.e. they are not restricted to Sphase when nuclear DNA replicates. In 1971 Jerome Vinogard and Piet Borst independently discovered
that mt DNA mols exhibit a small 100p.displaced from the main circle and mode of replication is semi
conservative. Denaturation of DNA mols causes a small fragment of S.S.DNA to be released, suggesting
that loop is caused by displacement of one of the two strand of double helix, therefore it is termed as D
loop (Displacement loop)
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mt DNA codes for ribosomal RNAs, t-RNAs and small mitochondrial polypeptides
Several evidence suppose the conclusion that mt DNA serves as a template for transcription of mt
RNAs. 2nd-mitocondria contains their own RNA-p-ase that can be distinguished from nuclear. RNAs
by differing susceptibility to inhibitors. 3rd ethidium bromide inhibits mt DNA synthesis and not nuclear
DNA synthesis supports the conclusion that independent RNA synthetic pathways exist in nuclear and
mitochondria.
mitochondria synthesis proteins using a unique sets of ribosomes, tRNAs and protein factors.
The mRNAs transcribed by mt DNA are translated into polypeptides within mitochondria using.
Protein synthesizing machinery that differs from the machinery present in the cytorol.
Ribosomes:
contraction (Table I)
I sizes of various types of ribosomes
Source
Mitochondria
Intact
Ribosomes
55-605
(mammals)
Mitochondria
(yeast)
755
Ribosomal
Sub units
Large 455
Ribosomal
RNAs
165
Small - 355
125
Large 535
215
Small - 355
145
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Mitochondria
785
(Plants)
Cytoplasm
805
Eukaryotes
2.2.2.2.
Large 605
265
Small 405
185, 55
Large 605
285
Small - 405
185, 5.85, 55
Transfer RNAs
Mitochondria contains t-RNAs and amino-acyl t-RNA synthetase. The amino-acyl t-RNA synthetase
are coded by nuclear DNA by t-RNAs are transcribed from mitochondria differs in sequence from the
corresponding t-RNAs found in cytosol. Mammalian mt DNA codes for only 22t RNAs following
wobble rules Another unusual features of mitochondrial t-RNA is presence of the t-RNA for
formylmethionine. This type of t-RNA is once thought to be found in bacterial only. Its presence in
mitochondria suggest that protein synthesis in the two organelle inhibits some similarities to the protein
synthesis of bacterial.
2.2.2.3
Protein synthesis factors of mitochondria are structurally distinct from its counterpart in the cytosol.
Several protein synthesis factors present in mitochondria are functionally inter changeable with those
obtained from bacterial cells.
2.2.2.4
Examining base sequence of mitochondrial DNAs has revealed that mRNA is translated into
polypeptide chair using a genetic code that is slightly different from universal code system difference in
codon usage can occur even among the mitochondria of different organism (Table II)
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Codon
Universal code
system
Mammals
Yeast
AUA
Ile
Met
Met
UGA
Stop
Trp
Trp
AGA,
Arg
Stop
Arg
Leu
Leu
Thr
AGG
4
CUU
CUC
CUA
CUG
2.2.3. BIOGENESIS
New mitochondria arises by growth and division of existing mitochondria or pro mitochondria
Microscopic evidences support this idea. Mitochondria in the process of dividing have been observed
under phase contrast microscope and membrane partition dividing mitochondria in half can be detected
by electron microscope in green algae. Micromonas which contain single mitochondria but the
microscopic observations do not rule out the possibilities that mitochondria also arise by other
mechanism. In 1960s David Luck designed experiment on Neurospora, that focused whether such
alternative mechanism exist. 1st
precursor), so it could be incorporated within mitochondrial membranes. Cultures were then transferred
to non-radioactive medium and allow to grow for varying periods of time and finally examined by
microscopic auto radiography and silver grains were counted to determine the distribution of
radioactivity one of the 3 patters would be emerged:
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1. If new mitochondria are formed by growth & division of existing mitochondria then each
doubling in the number of mitochondria, after the 3H choline is removed it should be
accompanied by 50 percent reduction in the no of auto radiographic grains/ mitochondria.
2. If de-novo synthesis occurs then new mitochondria formed after abnormal of 3H-choline should
be non-radioactive while the radioactivity in the pre-existing mitochondria would remain the
same.
3. If mitochondria develops from other membrane then the pre-existing radioactive mitochondria
should retain their radioactivity and new mitochondria should at 1st be radioactivity because the
membrane from which it develop would be radioactive. But as the time parse, the absence of 3Hcholine from the medium would decrease in the amount of radioactive in the cellular membranes
from which mitochondria develops and there form the average amount of radioactive/labeled
mitochondria would decrease Data obtained in the studies followed 1 st pattern suggesting that
new mitochondria aries from growth and division of existing mitochondria.
In some cases mitochondria develop in the cells that appears to have no pre-existing
mitochondria e.g. yeast which can survive either aerobic or aerobically. When switched to
anaerobic conditions, the cells meet all their energy requirement through glycolysis and so
mitochondria gradually disappears. When some cells are exposed to oxygen, mitochondria reappears. Previously such conditions was thought to be de-novo formation. But electron
microscopic examination revealed the presence of small double-membrane enclosed vesicular
about 1mm in diameter that lacks costae and cytochrome but contains mt DNA. When
anaerobically grown Yeast are exposed to-Oxygen the inner membrane forms crested that
synthesize new cytochrome and other respiratory chain components. Because of their ability to
differentiate into mitochondria the small double-membrane vesicles are called pro mitochondria.
It is bounded by two membrane with the inner membrane folded into cristae, that are responsible
for energy generation by the tricarboxylic acid cycle, electron transport and oxidative
phosporylation.
Within the region enclosed by the inner membrane, the matrix , they have DNA,ribosomes, and
other molecules necessary for protein synthesis
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Mitochondrial ribosomes are 70S type and posses altered genetic code system.
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2.2.6 ACTIVITIES/ASSIGNMENTS
Compare and list out the difference between plant and animal
mitochondria
2.2.8
REFRENCES-
2.3.0 VACUOLES
vacuoles plays several metabolic roles1. Storage and accumulation: vacuoles store a large varieties of molecules, including inorganic
ions, organic acids, sugars, enzymes, storage proteins and many types of secondary metabolites.
Proteins in the tonoplast transport all of these molecules except for storage proteins into the
vacuoles. The resulting accumulation of solutes in the vacuole drives the osmotic uptake of
water, producing the turgor pressure needed for cell enlargement. Some vacuoles have high
concentration of pigments, which produce the colour of many flowers or the red or red maple
leaves. In maple leaves red colour is so concentrated in epidermis that they mask the green
colour of the chloroplast many primary metabolites can be retrieved from vacuoles.
2. Plant cell expansion is driven by a combination of osmotic uptake of water into the vacuoles and
altered. Cell wall extensibility the water taken into the vacuoles generates turgor pressure which
not only expands the primary cell wall but also creates stiff load bearing structures in
conjunction with the wall. Wilting and the associated softening of plant organs is caused by loss
of water from the vacuoles and surrounding cytosol.
To maintain the turgor pressure of continuously expanding cells solutes must be actively
transported into the growing vacuoles to maintain its osmolarity. An electro chemical gradient
across the tonoplast provides the driving force for this uptake solutes. The gradient is produced
and maintained by two electrogenic protons pump. V-type H+ ATpase and vacuolar H+
pyrophosphates (H-Ppase). The principal solutes in vacuoles include the ions K+, Na+, ca2+,
Mg2+, Cl, So42, Po43 and No3 and primary metabolites, such as amino acids, organic acids and
sugar. The moment of water across the tonoplast is mediated by aquaporin channels consisting of
tonoplast intrinsic proteins (TIPs).
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Plants cannot escape from toxic sites nor they can eliminate toxic materials like heavy materials like
oxalate by excretion. Instead, plants sequester these compounds into vacuoles. For example to
remove oxalate specific cells develops vacuoles containing an organic matrix within which oxalate is
allowed to react with Ca to form Ca-oxalate crystals. In other plant cell types membrane of the ABC
family of transporters are used to transport xenobiotics from the cytoplasm into the vacuoles.
Accumulation of toxic compounds in leaf vacuoles is one of the reasons leaves are shed on a regular
basis.
2.3.2.5 Pigmentation
Vacuoles that contain anthocyanin pigments are found in many types of plant cells.
Pigmented flowers, petals and fruits are used to attract pollinators and seed disperse. Some leaf
pigments screen outs uv are visible light, preventing photo oxidation damage to the photosynthetic
apparatus. This screening appears to be essential for the survival of the leaves of evergreens that
grow in freezing conditions during hunters prevents the absorbed light energy being used in
photosynthesis.
They enables plant cells to grow rapidly with expenditure of minimal amount of energy .
They perform a multitude of functions including storage, digestion, ionic homeostasis, defence
against plant pathogens etc.
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mature cell
elongated cells
meristematic cells
b.
sugars
organic acid
soluble salts
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2.3.7 REFRENCES
BIOCHEMISTRY AND MOLECULAR BIOLOGY OF PLANTS BUCHANAN. GRUISSEM.
JONNES.
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