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fungi&#xd;
of&#xd;
the&#xd;
genus&#xd;
Epichloë&#xd;
are&#xd;
symbionts&#xd;
of&#xd;
cool&#xd;
season&#xd;
grasses.&#xd;
Epichloë&#xd;
infections&#xd;
are&#xd;
generally&#xd;
asymptomatic,&#xd;
with&#xd;
endophyte&#xd;
and&#xd;
host&#xd;
growth&#xd;
synchronised&#xd;
to&#xd;
form&#xd;
an&#xd;
interconnected&#xd;
hyphal&#xd;
network&#xd;
in&#xd;
the&#xd;
intercellular&#xd;
spaces&#xd;
separating&#xd;
host&#xd;
cells.&#xd;
In&#xd;
return&#xd;
for&#xd;
the&#xd;
sustenance&#xd;
received&#xd;
from&#xd;
the&#xd;
host&#xd;
apoplast,&#xd;
Epichloë&#xd;
spp.&#xd;
produce&#xd;
a&#xd;
range&#xd;
of&#xd;
bioprotective&#xd;
secondary&#xd;
metabolites&#xd;
(SM)&#xd;
that&#xd;
deter&#xd;
both&#xd;
vertebrate&#xd;
and&#xd;
invertebrate&#xd;
herbivores.&#xd;
Peramine&#xd;
is&#xd;
an&#xd;
Epichloë-­‐&#xd;
derived&#xd;
insect-­‐deterring&#xd;
SM&#xd;
produced&#xd;
by&#xd;
the&#xd;
two-­‐module&#xd;
non-­‐ribosomal&#xd;
peptide&#xd;
synthetase&#xd;
(NRPS)&#xd;
PerA,&#xd;
encoded&#xd;
by&#xd;
the&#xd;
gene&#xd;
perA.&#xd;
The&#xd;
perA&#xd;
gene&#xd;
is&#xd;
widespread&#xd;
across&#xd;
Epichloë,&#xd;
but&#xd;
peramine&#xd;
production&#xd;
is&#xd;
much&#xd;
more&#xd;
discontinuous.&#xd;
Through&#xd;
an&#xd;
Epichloë-­‐wide&#xd;
survey&#xd;
it&#xd;
is&#xd;
shown&#xd;
that&#xd;
the&#xd;
peramine&#xd;
chemotype&#xd;
of&#xd;
Epichloë&#xd;
isolates&#xd;
can&#xd;
be&#xd;
accurately&#xd;
predicted&#xd;
through&#xd;
a&#xd;
combination&#xd;
of&#xd;
PCR&#xd;
and&#xd;
sequencing&#xd;
methods.&#xd;
Furthermore,&#xd;
the&#xd;
protein&#xd;
encoded&#xd;
by&#xd;
the&#xd;
widespread&#xd;
peramine-­‐negative&#xd;
perA-­‐ΔR&#xd;
allele&#xd;
is&#xd;
analysed&#xd;
in&#xd;
detail.&#xd;
The&#xd;
perA-­‐ΔR&#xd;
allele&#xd;
has&#xd;
a&#xd;
transposable&#xd;
element&#xd;
inserted&#xd;
into&#xd;
the&#xd;
3’&#xd;
end&#xd;
of&#xd;
the&#xd;
gene,&#xd;
and&#xd;
was&#xd;
previously&#xd;
assumed&#xd;
to&#xd;
be&#xd;
non-­‐functional.&#xd;
Evidence&#xd;
for&#xd;
ongoing&#xd;
perA-­‐ΔR&#xd;
functionality&#xd;
is&#xd;
presented,&#xd;
with&#xd;
the&#xd;
PerA-­‐ΔR&#xd;
protein&#xd;
shown&#xd;
to&#xd;
exhibit&#xd;
altered&#xd;
substrate&#xd;
specificity&#xd;
relative&#xd;
to&#xd;
PerA.&#xd;
A&#xd;
PerA/PerA-­‐ΔR&#xd;
domain&#xd;
swap&#xd;
is&#xd;
shown&#xd;
to&#xd;
restore&#xd;
peramine&#xd;
production&#xd;
to&#xd;
PerA-­‐ΔR,&#xd;
providing&#xd;
insight&#xd;
into&#xd;
the&#xd;
functional&#xd;
changes&#xd;
that&#xd;
separate&#xd;
this&#xd;
protein&#xd;
from&#xd;
PerA.&#xd;
Another&#xd;
major&#xd;
project&#xd;
investigated&#xd;
the&#xd;
role&#xd;
SM&#xd;
genes&#xd;
may&#xd;
play&#xd;
in&#xd;
Epichloë&#xd;
sexual&#xd;
development.&#xd;
Many&#xd;
Epichloë&#xd;
spp.&#xd;
are&#xd;
transmitted&#xd;
vertically&#xd;
through&#xd;
infection&#xd;
of&#xd;
the&#xd;
host&#xd;
seed.&#xd;
However,&#xd;
sexual&#xd;
Epichloë&#xd;
spp.&#xd;
may&#xd;
also&#xd;
undergo&#xd;
a&#xd;
reproductive&#xd;
cycle&#xd;
that&#xd;
involves&#xd;
formation&#xd;
of&#xd;
pre-­‐sexual&#xd;
stromata&#xd;
structures&#xd;
over&#xd;
developing&#xd;
host&#xd;
inflorescences.&#xd;
This&#xd;
sexual&#xd;
cycle&#xd;
culminates&#xd;
in&#xd;
horizontal&#xd;
transfer&#xd;
of&#xd;
the&#xd;
endophyte&#xd;
to&#xd;
endophyte-­‐free&#xd;
host&#xd;
plants.&#xd;
Deletion&#xd;
of&#xd;
the&#xd;
symbiotically&#xd;
regulated&#xd;
endophyte&#xd;
gene&#xd;
irlA&#xd;
induced&#xd;
a&#xd;
symbiosis-­‐defective&#xd;
phenotype&#xd;
in&#xd;
planta,&#xd;
and&#xd;
this&#xd;
observation&#xd;
led&#xd;
to&#xd;
the&#xd;
identification&#xd;
of&#xd;
a&#xd;
novel&#xd;
Epichloë&#xd;
SM&#xd;
cluster.&#xd;
Synteny&#xd;
analysis&#xd;
and&#xd;
comparison&#xd;
to&#xd;
previously&#xd;
characterised&#xd;
fungal&#xd;
SM&#xd;
gene&#xd;
clusters&#xd;
defines&#xd;
a&#xd;
five-­‐gene&#xd;
cluster&#xd;
centred&#xd;
on&#xd;
irlA&#xd;
that&#xd;
is&#xd;
shown&#xd;
to&#xd;
be&#xd;
upregulated&#xd;
in&#xd;
stromata.&#xd;
A&#xd;
model&#xd;
is proposed&#xd;
whereby&#xd;
this&#xd;
SM&#xd;
cluster&#xd;
controls&#xd;
initiation&#xd;
of&#xd;
the&#xd;
proliferative&#xd;
hyphal&#xd;
growth&#xd;
associated&#xd;
with&#xd;
stromata&#xd;
formation.</mods:abstract>
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               <mods:accessCondition type="useAndReproduction">The Author</mods:accessCondition>
               <mods:subject>
                  <mods:topic>Epichloe</mods:topic>
               </mods:subject>
               <mods:subject>
                  <mods:topic>Genetics</mods:topic>
               </mods:subject>
               <mods:titleInfo>
                  <mods:title>Characterisation of novel secondary metabolism genes in plant-endophytic Epichloë fungi : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Genetics at Massey University, Manawatu, New Zealand</mods:title>
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