UDAI PRATAP AUTONOMOUS

COLLEGE,VARANASI
TOPIC-
HETEROCYST DIFFERENTIATION

SUBMITTED TO- SUBMITTED BY-

Dr. ARVIND PRATAP SINGH ASHUTOSH KUMAR SINGH
CONTENTS-
1. Introduction of heterocysts.
2. Nitrogenase genes and Neighbouring
DNA.
3. Electron Donor to Nitrogenase.
4. Organic electron donors.
5. Intercellular movement of carbon
compounds.
6. PS-I and PS-II.
7. Relationship of diverse differentiation
processes in cyanobacteria.
Introduction of Heterocysts-
 Perhaps 2.2 Billion years ago, our world was
anaerobic and autotrophic organism that could
gain their energy from sunlight, their carbon and
(often) their nitrogen from the atmosphere and
their reductant from water were in the
ascendency as they grew and flourish they
polluted the world with anoxious byproduct,
OXYGEN , derived from photoxidation of water.
 Although inhibiting too many biochemical
reactions, oxygen was shelldown as detrimental
as it was to be process of nitrogen fixation,
 Because of the exceptionally negative reducing
potentials required for that process.
 Some of these autotrophic microorganisms
adopted to the newly changed environment by
making there Nitrogen fixing
enzymes(Nitrogenase) only when oxygen
happened to be absent.
 Others such microorganisms found that
although it was necessary to stop nitrogen
fixation during the day when there
intracellular oxygen concentration was lowest.
 Yet others make the challenge of increased
atmospheric oxygen by modifying a small
percentage of their cells for the task of fixing
nitrogen. In the last of these rows physiological
differentiation was accompanied by
morphological differentiation.
 The nitrogen fixing cells are known as
heterocysts from the time when there function
was unknown.
 The heterocyst provide the nitrogen that is
needed by the organism living to the other
cells.
 Heterocyst, whose essential nature
involves metabolic exchange, evolved in
filamentous cyanobacteria.
 They are unknown in unicellular
cyanobacteria and have not been reported
to differentiate in unicellular mutants of
filamentous species.
 Optimal proliferation when fixing
nitrogen is limiting requires co-ordination
of the differentiation of the some
vegetative cells into fixing heterocysts.
 While others continue to fix co2 and
replicate, it remains a challenge to
elucidate how it is determined which
vegetative cells are to differentiated into
heterocysts and how they do so.
 In order to avoid inactivation of
nitrogenase by oxygen, heterocysts stop
synthesizing oxygen and limit the rate of
entry of that gas presumably because the
vanderwalls radii of N2 AND O2 are
similar, 1.5Å and 1.4Å respectively.
 Evolution apparently found no way to
permit to enter while excluding O2.
however a barrier that consists of laminae
of long chain lipid molecules and that is
surrounded in term by a protective layer
of polysaccharides, reduces the
permeability of both gases to the extent
that into enters still suffices for the need
of organism for nitrogen.
 While the O2 that enters can be reduced
to water by respiration.
 Because photosynthetic water splitting,
the ultimate source of electrons used for
N2 fixation, cannot continue in
heterocysts.
 It must be replaced by movement of
reductant from(photosynthesis)
vegetative cells to heterocysts.
 In addition, there must be movement of
fixed nitrogen from heterocysts to
vegetative cells, therefore some channel
must remain open between the two types
of cell.
 Although entry of O2 around the periphery of
the heterosis can be largely eliminated.
 While the envelope layers, no such barriers
may be present in the polar region, in order not
to block the channel was movement of
nutrients.
 Such channel should therefore have a cross-
sectional area no longer than required to
support the needed flux of nutrients; and
respiration especially next to those channel is
required to maintain microaerobiosis.
 Within the heterocysts, infact a
‘Honeycomb’ of membranes rich in
oxidative enzymes is located in the
region of the heterocysts near the
channels.
 Nature having an aerobic world, one
wonders to what uses it may be put in
addition to nitrogen fixation.
Nitrogenase genes and Neighbouring DNA-
 To fix into and to maintain the capacity
under anaerobic conditions.
 Heterocysts required the product of nif
genes, reductant and ATP.
 nitrogen fixation is catalyzed by
dinitrogenase a dimer of the polypeptides
encoded by nif D and nif K.
 Which is supplied with electrons by
dinitrogenase reductase, which is a dimer
of the polypeptide encoded by nif H.
 These genes (nif H, nif B, nif K) are contagious
and formed an operon in the chromosomes of
heterocysts.
 Dinitrogen reductase containing a single 4Fe-
4S. Cluster that can be irreversibly oxidise by
O2.
 Dinitrogenase contains 4Fe-4S centers that are
arranged in two clusters and two copies of
FeMo-Co, which is an iron and molybdenum
containing co-factor all of which add to the
oxygen ability of the enzyme.
 Dinitrogenase and dinitrogenase reductase
together comprisers nitrogenase.
 The nitrogenases of various diazotrophic
organisms exhibit a high degree of sequence
similarity at the amino acid level and often on
the DNA level.
 The nitrogenase of heterocyst may therefore
resembles in its 3-Dimensional structure,
models constructed from x-ray,
crystallographic data of azotobacter vinelandii
nitrogenase.
Electron Donor to Nitrogenase-
 Two plants types 2Fe-2S ferrodoxins were
found in heterocyst of Anabaena variabilis,
found only in heterocyst is encoded by the
gene fdxh, while the other is similar and
perhaps identical, to the ferredoxin observed
in vegetative cells and encoded by PET F.
 The product of translation of the two genes
differ in 47 out of 98 amino acids
 The occurrence of an additional, bacterial type
ferodoxins in Anabaena variabilis was recently
reported, it may possibly be encoded by fdxn.
Organic electron donors-
Organic electron donors-
 Because photosynthetic water splitting in
heterocyst would be incompatible with
nitrogenase activity it appears highly
likely that the reduction of N2 and O2.
 In heterocyst is supported by organic
metabolites from vegetative cells
alternatives into reduce inorganic solutes
such as S2-, reduce proteins or solid state
transfer via membranes,
 But there is no substantive support for
ending of these possibilities a vairiety of
metabolites can be oxidised by
dehydrogenases with concommitant
reduction of NADP+ or NAD+.
 The NADPH that results can reduce
ferredoxin or transfer electrons to PS-1 or
the respiratory electron transport chain in
each case via ferredoxin: NADP+ and
oxidoreductase whereas the NADH that
results can transfer electrons to PS-1 or to
the respiratory electron transport chain.
 The 3 metabolites are the primary candidate
for electron donor to NADP+ in heterocyst are
glucose-6-phosphate and 6-
phosphoglutonate(both of which are oxidised
by the oxidative pentose phosphate cycle,
 Reduction of NADP+ is enzymatically coupled
to the oxidation of glyceraldehyde-3-phosphate
and malic by the corresponding
dehydrogenases.
 In addition, the pyruvate has the capacity to
reduce ferrodoxin directly in a reaction
catalysed by pyruvate.
 Ferrodoxin isoreductase enzymes of the
oxidative pentose phosphate cycle are
highly active in the heterocysts.
 Whether the triosephosphate produced by
the oxidative pentose phosphate cycle can
be furthur metabolised by glycolysis
within heterocyst depends upon the
availability of glyceraldehyde-3-P-
dehydrogenase. That enzyme whose
availability was previously controversial
appears now to be abundantly present in
heterocysts of anabaena cylindrica.
 Anabaena variabilis, and anabaena spp.
Strain PCC7119 where it can also serve
as a source of NADH and of phospho-
enol-pyruvate(PEP).
Intercellular movement of carbon
compound-
 Pulse chase experiment demonstrated
unimplivocally that 14C from 14CO2 Fixed
by vegetative cells moves into heterocysts
and is metabolised there whatever the
carbon containing substances is (or) that
move into heterocyst they presumably are
a source of electron for nitrogen fixation
of carbon skeletons, for assimilation of
fixed nitrogen and of building blocks for
envelope materials in the heterocysts.
Photosystem-I, Source of ATP and
Reductant-
 Heterocysts contain PS-I.
 In several strains of Anabaena and Nostoc
species, the ratio of total chlorophyll per
PS-I reaction centre in heterocysts is 1\3 to
2\3 as great as in vegetative cells.
 PS-I driven photophosphorelation could
involve cyclic or linear electron flow,
Do heterocysts have PS-II?
 During the differentiation of heterocyst
O2 evolution is lost and pigment
associated with PS-II may be lost.
 Although the O2 sensitivity of nitrogenase
leads to the expectation that heterocysts
would not produce O2 in spite of that less
retain a modified PS-II i.e, able to receive
electrons from an organic or (inorganic)
donor.
 Whether a high potential form of
cytochrome b559, a component of the
PS-II reaction centre is present in
heterocyst wascontroversial a decade
ago and remains controversial.
Relationship of diverse differentiation
processes in cyanobacteria-
 Several observations support the hypothesis
that the differentiation of the heterocysts and
the differentiation of akinetes are related
processes.

1. The envelopes of akinetes(spores) of

Anabaena cylindrica and Anabaena variablis
have the same polysaccharide that is
characteristic of the envelopes of the
heterocysts of the same species.
2. Akinetes and heterocysts are both
non-dividing cells.

3. In certain taxa, akinetes often or

occasionally form where the
heterocysts would be expected to
form.