Physiology of

Microorganisms
(B 419)
4th Level Chemistry-Botany
Students
Ass. Prof. Dr. Mervat H. Hussein
Botany Dept
Faculty of Science
Mansoura University
Apical Growth
Mechanism
In Fungi
 Apical growth
• It is the apical extension,
because the true growth,
defined as an increase in
biomass per unit time
 Ultrastructure of Hyphae
• A- Young region of a hyphae
• B- Part of a mature region of a hyphae
• C- Close-up of the Spitzenkŏrper
• i.e. an accumulation of small,
membrane-bound vesicle of
different sizes & contents,
surrounding a central, vesicle-
free core
 Vesicles of the APICAL VESICULAR
CLUSTER (AVC)
Spitzenkŏrper
• Wall Precursors - the sub-units
or building blocks of the wall
polymers - e.g. uridine diphosphate
N-acetylglucosamine, the sub-unit
of chitin
• Wall Lytic Enzymes - which help
breakdown and separate wall
components - e.g. chitinase,
glucanase
• Wall Synthase Enzyme - which help
assemble new wall components and
so increase the size of the wall - e.g.
chitin synthase, glucan synthase.
 Apical Growth
• *Fungi are unique in that growth occurs
solely at the apical tip of a hypha
• *Hyphal tip (apex) shows extreme
plasticity
• Swell to form spores or yeast-like cells
• Taper to penetrate
• Give rise to complex structures
 Model 1 - involvement of wall
lytic enzymes:
• According to this model, if the hypha is
going to be able to extend at its tip,
there will have to be:
• some softening (lysis) of the existing
wall, and
• the synthesis and incorporation of
new wall material.
 1. Vesicles containing lytic enzymes or wall
precursors move through the cytoplasm
towards the hyphal tip, where they fuse with
the plasma membrane, releasing their contents
into the wall
2. The lytic enzymes released into the wall
attack the polymeric fibrils.
3. The weakened fibrils stretch out and become
separated from one another due to the turgor
pressure of the protoplasm.
4. Synthase enzymes and wall precursors build
new fibrils and synthesize additional
amorphous components of the wall.
5. The surface area of the hyphal wall increases.
Fusion of the vesicles with the plasma
membrane ensures that the former contribute to
the increase in surface area of the latter.
 Assembly of the wall at the hyphal
apex
• Wall synthesis at the hyphal apex is
a complex process includes:
• 1- Chitin Synthase
• 2- Glucan synthase
• 3- Mannoproteins
• 4- Cross-linking & maturation of the
hyphal wall
• 5- Wall lytic enzymes
 Chitin Synthesis
• Chitin synthase catalysis the
synthesis of chitin chains, and is
therefore one of the principal
enzymes involved in fungal wall
synthesis.
 1- Chitin Synthase
• The substrate for chitin synthesis is N-
acetylglucoseamine, where it is supplied
as a sugar nucleoid, UDP-N-
acetylglucoseamine
(UDP= uridine diphosphate), with a
high-energy bond required for chitin
synthesis.
 Chitin synthase - biosynthesis of
chitin
• Chitin is formed in situ, not delivered by
membrane-bound vesicles
• Chitin synthase is probably delivered by
vesicles containing chitosomes which are
inactive zymogens
• Chitin synthase becomes an integral
membrane protein that must be activated by a
protease (also probably delivered by vesicles)
• Chitin fibrils are synthesized to
extrude from the outer face of the
plasma membrane
• The synthesis of chitin is regulated
by many mechanisms including a
cytosolic inhibitor
 2- Glucan synthase
• It catalyses the synthesis of β- 1,3-glucan
chains, which often comprise the bulk of
the fungal wall.
• Glucan synthase arrive in vesicles and
becomes inserted in the plasma membrane
at the apex.
• The substrate is a sugar nucleotide (UDP-
glucose), supplied from the cytosol.

•
 Glucan synthase - produces b-1,3-
glucan
• Probably delivered by vesicles and
incorporated into the plasma membrane in
a manner similar to that of chitin synthase
* Regulation differs - two subunit enzyme
• Catalytic subunit on outside portion of
membrane
• GTP-binding protein subunit on inside of
membrane
• GTP binding stimulates the synthesis of
glucan
• Modifications of glucans by the
addition of side chains of b-1,3-
glucan which progressively increase
in the subapical hyphae regions
during wall maturation
 3- Mannoproteins
• Mannoproteins form a relatively
small proportion of the fungal
hyphae, but are more common in
yeasts & in dimorphic fungi
• It is preformed in the endoplasmic
reticulum-Golgi complex & derived
to the apex in vesicles
• Mannoproteins are pre-formed
in the endoplasmic reticulum-
Golgi complex and delivered to
the apex via vesicles; mainly a
major component of yeast cell
walls
 4- Cross-linking & maturation of the
hyphal wall
• Pure glucans can be complexed with
chitin & and can be associated with one
another.
• These additional bondings behind the
growing apex could serve to convert the
initially plastic wall into a progressively
more cross-linked & rigidified structure
• Cross-linking and maturation -
various linkages between wall
components occur more
regularly in the older parts of the
wall
*lysine may be involved in some
linkages
 5- Wall lytic enzymes
• One point of view:
• The existing wall must be softened in
order for new wall components to be
inserted, as wall growth would involve a
balance of wall lysis and wall synthesis.
• Chitinase, Cellulase (in Oomycetes),
& β- 1,3-glucanase activities can be
found
• Other point of view:
• The substantial cytoskeleton of tubulins
& actin could help to reinforce the
hyphal tip, precluding the need for a
rigid wall & therefore precluding the
need for wall-depending enzymes
• Production of new tips (new yphal
branches)
 Wall lytic enzymes - chitinase,
glucanase
• May be involved in the “softening” of
the wall at the apical tip, though some
evidence suggests that it is the
cytoskeleton that reinforces the hyphal
tip
• Must be required for initiation of
branching at subapical regions of the
hypha
Simplified model of apical growth of fungal
hyphae. G = Golgi body; V = vesicle; M =
microtubule
 Model 2 - steady state:
• lytic enzymes are NOT involved in apical
growth
• because the newly formed wall at the extreme
tip of the hypha is Viscoelastic (essentially
fluid)
• so that as new wall components are added at
the tip, the wall flows outwards and
backwards (diagram after…)
• and the wall then Rigidifies progressively
behind the tip by the formation of extra
chemical bonds
• New wall polymers synthesized
at the extreme tip are suggested
to flow outwards and backwards
as new components are
continually added at the tip.
 The driving force for apical growth
• Cytoskeletal components
• Saprolegnia (Oomycetes): the apex can
extend even when hyphae have
negligible turgor pressure…….. Actin
polymerization
• Actin is abundant in hyphal tips
• Tip extension & cytoplasmic streaming
can be halted by treating fungi with
Cytohalatins (cell-relaxers) which bind to
actin.
Involvement of Microtubules in fungal tip
growth
• Hyphal extension can be halted by
benzimidazole fungicide & the related
azole drugs, that all of which interfere
with microtubule function as there are a
progressive depletion of vesicles in the
hyphal tip ……..
• Microtubules may provide Tramlines
for vesicle cargoes
 Steady-state model of wall
growth
• Proposed by Wessels (1990)
• Newly-formed wall is viscoelastic
• Wall polymers continually added to the
tip
• Flow outwards and backwards Wall
rigidifies progressively
• Calcium may be involved in tip fungal
growth as:
• 1- it required in external supply for
growth,
• 2- the plasmalemma at the extreme tip is
reported to have a high concentration of
stretch-activated calcium channels,
allowing the ingress of calcium when the
membrane is stretched…….
• Calcium cause the contraction of F-actin
 The Cell Envelope
• Ergosterol is the major sterol of p.
m. in fungi
• A macromolecule coating …
Glycocalyx outside the p. m.
 Definition of Growth in Fungi
• Growth may be defined as an
irreversible increase in the volume of an
organism, usually accompanied by an
increase in biomass.
• Mycelial fungi exhibit extension growth
of hyphae, accompanied by an increase
in biomass.
• Unicellular fungi (e.g. yeasts) may
exhibit an increase in individual cell
volume, accompanied by an increase in
biomass.
• But collectively, the number of yeast
cells within a culture (i.e. cell
concentration) may also increase,
resulting in an increase in biomass of the
culture as a whole.
• Fungi may be cultured on SOLID or in
LIQUID MEDIA:
• YEASTS are often cultured in LIQUID
media
 Growth Kinetics in Liquid
Media: Batch Culture

• If we plotted biomass or cell

concentration against time we might
obtain the following characteristic S-
shaped growth curve
1. Characteristic S-shaped growth curve:
• During an initial Lag phase the rate
of growth or cell division is very
slow.
• Growth or cell division then starts to
accelerate into the EXPONENTIAL
phase
• Growth will start to DECELERATE
(DECLINE).
• This may be followed by a STATIONARY
phase, during which there is no
discernible change in cell concentration
or biomass.
• Finally, we may observe a phase of CELL
DEATH and LYSIS - which results in a
decrease in cell number and/or
biomass.
 The Maintenance of Polarity
• Polarized transport of vesicles to the apex.
• Onward growth:
The progressive strengthening and
thickening of the wall by cross-
linking of chitin and glucan
components and addition of
materials
 The Fungi
)‫(مكتبة الكلية‬
• Michael J. Carlile,
& Sarah C. Watkinson, 1998
• Bot. 889
• 17527
• *P. 84- 86
• * P. 94 – 100
• * P. 102 - 109
• * Fungal Vegetative growth in Fungi
• The Growth of Populations and Colonies