REVIEW
Conidia ion in Neu ospo a c assa: ege a i e ep oduc ion
by a model ungus
Ca men Ruge -He e os
1,2
&Luis M. Co ochano
3
Recei ed: 12 Ma ch 2019 /Re ised: 14 May 2019 /Accep ed: 20 May 2019
#Sp inge Na u e Swi ze land AG 2019
Abs ac
Asexual de elopmen , conidia ion, in he ilamen ous ungus Neu ospo a c assa is a simple de elopmen al p ocess ha s a s
wi h he g ow h o ae ial hyphae. Then, he o ma ion o cons ic ions and subsequen ma u a ion gi es ise o he ma u e conidia
ha a e easily dispe sed by ai cu en s. Conidia ion is egula ed by en i onmen al ac o s such as ligh , ae a ion and nu ien
limi a ion, and by he ci cadian clock. Di e en egula o y p o eins ac ing a di e en s ages o conidia ion ha e been desc ibed.
The ole o ansc ip ion ac o s such as FL, and componen s o signal ansduc ion pa hways such as he cAMP phosphodies-
e ase ACON-2 sugges a complex in e play be ween di e en ial ansc ip ion and signal ansduc ion pa hways. Compa isons
be ween he molecula basis o conidia ion in N. c assa and o he ilamen ous ungi will help o iden i y common egula o y
elemen s.
Keywo ds Asexual de elopmen .Conidia ion .Neu ospo a .Spo ula ion
In oduc ion
Neu ospo a c assa is an ascomyce e he e o hallic ilamen ous
ungus ha g ows as b anched mul inuclea ed hyphae wi h
pe o a ed sep a. In na u e, species o Neu ospo a ha e been
ound in a wide ange o a eas ha include opical, sub op-
ical and empe a e egions, and he ungus is easily de ec ed
g owing on he su ace o i e-sco ched ege a ion because o
i s abili y o me abolize cellulose and he ac i a ion by hea o
he sexual spo es (ascospo es) (Jacobson e al. 2006;Jacobson
e al. 2004; Luque e al. 2012; Tu ne e al. 2001). N. c assa
has been used as a model o ganism o he esea ch on se e al
aspec s o euka yo ic biology, including he mechanism o
ecombina ion, genome de ence by RNAi, ci cadian clock
egula ion and ligh sensing (Da is and Pe kins 2002;
Pe kins and Da is 2000; Roche e al. 2014) The li e cycle o
N. c assa includes asexual ep oduc ion and he de elopmen
o ege a i e conidia ha a e easy o dispe se. The de elop-
men al p ocesses ha lead o conidia ion in N. c assa a e,
howe e , e y di e en om hose egula ing conidia ion in
Aspe gillus nidulans, despi e bo h being membe s o he
Ascomyco a. He e, we e iew ou knowledge o he egula-
ion, he gene ics and molecula basis o conidia ion in
N. c assa. We p opose ha he simpli ied mode o ege a i e
ep oduc ion in N. c assa and ela ed ungi may help o un-
de s and asexual de elopmen in o he ascomyce es wi h mo e
complex conidia ion pa hways.
Mo phological e en s du ing conidia ion
in Neu ospo a c assa
The e a e wo pa hways o asexual ep oduc ion in N. c assa:
mac oconidia ion and mic oconidia ion. Mac oconidia ion
(hence o h conidia ion) is induced by he ans e om a liq-
uid o an ai in e ace, desicca ion and nu ien deple ion, and
is in luenced by se e al en i onmen al ac o s such as ligh
and CO
2
le els, and by he ci cadian hy hm (Sp inge 1993).
Conidia ion is a e y simple de elopmen al p ocess. The in-
duc ion o conidia ion p omo es a change in he di ec ion o
g ow h o he ege a i e hyphae ha g ow away om he
subs a e, and leads o he o ma ion o a mass o ae ial
*Ca men Ruge -He e os
c. uge -he e [email protected] g.de
1
Cen e o Molecula Biology o he Uni e si y o Heidelbe g
(ZMBH), (DKFZ-ZMBH Alliance), Im Neuenheime Feld 282,
69120 Heidelbe g, Ge many
2
Ge man Cance Resea ch Cen e (DKFZ), Im Neuenheime Feld
280, 69120 Heidelbe g, Ge many
3
Depa amen o de Gené ica, Facul ad de Biología, Uni e sidad de
Se illa, Apa ado 1095, 41080 Se illa, Spain
In e na ional Mic obiology
h ps://doi.o g/10.1007/s10123-019-00085-1
hyphae. Abou 4 h a e conidial induc ion, hyphal g ow h
changes om apical elonga ion o apical budding leading o
he o ma ion o chains o p oconidia ha a e di ided by mi-
no cons ic ions. Budding con inues in p oconidial chains,
and majo cons ic ions appea app oxima ely 8 h a e he
induc ion o conidia ion. This esul s in he sepa a ion o each
p oconidia du ing hei ma u a ion in o ull conidia (Fig. 1A).
In e conidial junc ions a e clea ed se e al hou s la e , bu
agile connec i e h eads hold conidia oge he un il hey
a e dispe sed by wind cu en s (Sp inge 1993; Sp inge and
Yano sky 1989). The en i e p ocess o conidia ion akes be-
ween 12 and 24 h. Howe e , conidia unde go a biochemical
ma u a ion pe iod o se e al days be o e hey a e able o e i-
cien ge mina ion (Fig. 1B).
The o he asexual ep oduc ion pa hway, mic oconidia ion,
is o en obse ed in old cul u es. Mic oconidia a e
uninuclea ed spo es mo phologically and de elopmen ally
di e en om mac oconidia. I has been shown ha mu an s
a ec ed in mac oconidia ion a e no a ec ed in
mic oconidia ion, sugges ing ha bo h pa hways a e indepen-
den and should ha e ew common egula o y elemen s, i any
(Maheshwa i 1999). Mic oconidia eme ge om a p o ube -
ance in he ege a i e hyphae which cons ic s un il
mic oconidia a e libe a ed. Ex e nally, mic oconidia a e
smalle han conidia and do no con ain ca o enoids in hei
cell wall. They a e less iable han mac oconidia (Sp inge
1993;Sp inge e al.1992; Sp inge and Yano sky 1989). In
addi ion, mic ocycle conidia ion has been obse ed in wild
ype isola es o N. c assa, bu he gene ics and mo phological
cha ac e iza ion ha e been limi ed (Maheshwa i 1991).
The egula ion o conidia ion
The p ocess o conidia ion is egula ed by a numbe o en i-
onmen al ac o s and endogenous signals ha should be p op-
e ly coo dina ed by he ungus in o de o p oduce conidia
when en i onmen al condi ions a e app op ia e o ege a i e
ep oduc ion. Conidia o ma ion is no mally obse ed when
he ungus is exposed o ai , bu ca bon s a a ion can induce
conidia ion in subme ged mycelia (Madi e al. 1994; Madi
e al. 1997). In N. c assa and o he ungi, de elopmen al cy-
cles a e o en ini ia ed by he same en i onmen al s esses ha
induce he accumula ion o high le els o ehalose, including
ca bon s a a ion, inc eased empe a u e and desicca ion (Li
e al. 1997; Sa gen and Kal enbo n 1972; Sp inge and
Yano sky 1992; Sun e al. 2011). Mo eo e , he p ocess o
de elopmen i sel is conside ed o cause physiological s ess
on o ganisms. In ma u e conidia, highe le els o ehalose
and s ess esponse p o eins may be equi ed o esis ance
and su i al (Hä ke e al. 1998; Rensing and Monne jahn
1998). Thus, ehalose may play a dual ole in he cell, unc-
ioning as a ese e ca bohyd a e o u u e hyphal g ow h and
Fig. 1 (A) De elopmen al
imeline o conidia ion in
Neu ospo a c assa. (B) Di e en
s ages o conidial de elopmen
isualized by scanning elec on
mic oscopy (a, b) and luo es-
cence mic oscopy (c, d, e, ): his-
one H1-GFP o nuclei isuali-
za ion and calco luo whi e o
cell wall s aining. A ows indica e
majo and mino cons ic ions (b),
hyphal sep a (a, e) and
in e conidial junc ions ( )
In Mic obiol
as a s ess p o ec an . The mechanism o induc ion o
conidia ion a e he ansi ion o an ai in e phase is no well
unde s ood bu i has been p oposed ha he de elopmen o
conidia helps he ungus o cope wi h he excess o damaging
oxygen adicals (Hansbe g e al. 1993; Pe aza and Hansbe g
2002).
Ligh egula es conidia ion (Sp inge 1993), and he egu-
la ion by ligh o conidia ion equi es he ac i i y o p o eins
WC-1 and WC-2 (Lau e and Russo 1990). WC-1 con ains a
zinc inge , a ch omopho e-binding domain (LOV) and PAS
domains o p o ein-p o ein in e ac ions (Balla io e al. 1996;
C os hwai e e al. 1997). The ch omopho e-binding domain
binds he la in FAD, allowing WC-1 o ac as a blue-ligh
pho o ecep o (F oehlich e al. 2002; He e al. 2002). The
p o ein WC-2 con ains a zinc inge and a PAS domain, and
in e ac s wi h WC-1 (Linden and Macino 1997) o o m a
Whi e Colla Complex (WCC). This complex, upon ligh ex-
posu e, binds ansien ly o he p omo e s o ligh -inducible
genes o ac i a e hei ansc ip ion (Belden e al. 2007;
F oehlich e al. 2002;HeandLiu2005; Smi h e al. 2010),
including he p omo e o l ( lu y), a gene ha encodes a key
egula o o conidia ion. Conidia ion inc eases in N. c assa
cul u es exposed o ligh sugges ing ha ligh may ac i a e
he ansc ip ion o key egula o y genes which indeed accu-
mula e a e ligh exposu e as i has been shown o l o csp-1
among o he s (Belden e al. 2007;Chene al.2009; Olmedo
e al. 2010a;Olmedoe al.2010b).
Conidia ion is egula ed by he endogenous ci cadian clock
esul ing in bands o conidia e e y 22.5 h o g ow h in he
da k (Dunlap and Lo os 2017). The mechanism by which he
clock egula es conidia ion has been cha ac e ized in de ail
and is based on a nega i e eedback loop in which he posi i e
elemen is he WCC which ac i a es ansc ip ion o q.FRQ
is he nega i e elemen o he clock and oge he wi h i s
pa ne s he RNA helicase FRH and he casein kinase-1
(CK-1) blocks i s own ansc ip ion as well as he ansc ip ion
o he o he WCC- egula ed genes (C os hwai e e al. 1997;
Dunlap and Lo os 2004; Hein zen and Liu 2007). The oscil-
la o y amoun o FRQ du ing he ci cadian cycle p omo es
changes in he abundance, phospho yla ion and ac i i y o
he WCC du ing g ow h in he da k (B unne and
Scha meie 2006;Dunlap2006; Dunlap and Lo os 2006).
Gene ics o conidia ion: egula o y genes
and p o eins
Gene ics has helped o iden i y some o he key elemen s ha
egula e conidia ion in N. c assa. Se e al genes equi ed o
conidia ion ha e been desc ibed and hei gene ic in e ac ions
ha e been es ablished. S ains wi h mu a ions in aconidia e-2
(acon-2)o lu yoid ( ld) a e blocked in he ansi ion om
ilamen ous o budding g ow h. Mu a ions in aconidia e-3
(acon-3)o lu y ( l) allow he p oduc ion o mino
cons ic ions, bu de elopmen is blocked and ew majo con-
s ic ions be ween p oconidia a e de ec ed. Mu a ions in wo
conidial sepa a ion genes (csp-1 and csp-2) p e en he sepa-
a ion o he c oss walls in ma u ed conidia o elease ee
conidia (Sp inge 1993). O hese genes, a en ion has ocused
on he p oduc s o genes l (NCU08726), csp-1 (NCU02713)
and csp-2 (NCU06095) ha a e ansc ip ion ac o s ha
should ac by egula ing ansc ip ion o genes ha pa ic-
ipa e in he de elopmen o conidia. These genes a e all
ligh -inducible p o iding a connec ion be ween ligh eg-
ula ion and conidia ion (Bailey and Ebbole 1998;Chen
e al. 2009; Lamb egh s e al. 2009;Pa ée al.2012;
Smi h e al. 2010).
The l gene has been cha ac e ized in de ail. The FL p o ein
is a 792-amino acid polypep ide con aining a Zn
2
Cys
6
binuclea zinc clus e domain belonging o he Gal4p amily
(Bailey and Ebbole 1998). Mu a ions in l block conidia ion a
he o ma ion o mino cons ic ions, ew hou s a e he in-
duc ion o conidia ion (Sp inge and Yano sky 1989). l
mRNA accumula es in ae ial hyphae whe e conidia ion-
speci ic genes a e exp essed sugges ing an impo an ole o
FL in he exp ession o hose genes (Bailey-Sh ode and
Ebbole 2004). Howe e , i has been also ound ha l
mRNA accumula es sho ly a e induc ion o conidia ion sug-
ges ing an addi ional ole o FL in he o ma ion o ae ial
hyphae (Co ea and Bell-Pede sen 2002). The impo ance o
FL as one o he majo egula o s o conidia ion in N. c assa
elies on he obse a ion o conidial de elopmen when l is
o e exp essed in subme ged ege a i e hyphae (Bailey-
Sh ode and Ebbole 2004). The o e exp ession o l in ege a-
i e hyphae leads o he exp ession o eas (Bailey-Sh ode and
Ebbole 2004), he gene o he hyd ophobin odle p o ein
loca ed on he su ace o ma u ed conidia (Bell-Pede sen
e al. 1992; Lau e e al. 1992). This obse a ion suppo s
he desc ibed binding o FL o he eas p omo e
(Re ngsam an e al. 2005). O he egula o y genes ha e been
ound o be up egula ed when l is o e exp essed in ege a i e
mycelia, including he conidia ion-speci ic genes con-6 and
con-10 (Re ngsam an e al. 2005) suppo ing he p oposal o
FL as a conidia ion-speci ic ansc ip ion ac o . I has been
desc ibed ha he aconidial pheno ype o a l mu an can be
pa ially supp essed by mu a ion in he gene ib-1, a ansc ip-
ion ac o ha is in ol ed in he egula ion o he e oka yon
incompa ibili y. This would sugges ha FL may egula e
conidia ion h ough he ep ession o VIB-1 (Xiang and
Glass 2002). The l gene is di ec ly ac i a ed by ligh h ough
he binding o he WCC o a ligh egula o y elemen on i s
p omo e loca ed a posi ion −640 om ini ia o ATG
(Olmedo e al. 2010a). l mRNA accumula es hy hmically
in an ACON-2-dependen manne (Co ea and Bell-
Pede sen 2002), suppo ing he p oposal ha he hy hmic
p oduc ion o conidial bands equi es he hy hmic accumula-
ion o l mRNA.
In Mic obiol
O he conidia ion mu an s ha e been isola ed and cha ac-
e ized. ACON-2 is a cAMP phosphodies e ase, bu i s ole in
he egula ion o conidia ion emains o be in es iga ed in
de ail (G eenwald e al. 2010), and FLD has been p oposed
o be a ansc ip ion ac o (Ca illo e al. 2017;McCluskey
e al. 2011). ACON-3 is a p o ein wi hou any iden i ied do-
main and is he homologue o MedA, a p o ein ha pa ici-
pa es in he egula ion o conidia ion o Aspe gillus nidulans
(Chung e al. 2011). I is in e es ing o no e he abundance o
ansc ip ion ac o mu an s ha a e blocked in conidia ion.
CSP-1 is a ligh -inducible zinc inge ansc ip ion ac o
(Lamb egh s e al. 2009; Smi h e al. 2010) and CSP-2 was
iden i ied as a ligh - egula ed g ainy head-like ansc ip ion
ac o (Colo e al. 2006;Pa ée al.2012). CSP-2 is in ol ed
in he de elopmen and emodelling o he cell wall and plays
a ole in he ac i a ion o genes ela ed wi h de ence and
i ulence (Pa é e al. 2012). A hi d ansc ip ion ac o ,
FLB-3, has been ecen ly cha ac e ized. I is he N. c assa
homologue o FlbC, a ansc ip ion ac o ha pa icipa es in
he egula ion o A. nidulans conidia ion. The lb-3 mu an is
blocked in conidia ion, shows al e ed sexual de elopmen and
is unable o comple e he sexual cycle (Boni e al. 2018).
These esul s sugges ha FLB-3 plays a key ole in he an-
sc ip ional coo dina ion be ween asexual and sexual
ep oduc ion.
T ansc ip ional egula ion du ing conidia ion
The disco e y ha se e al ansc ip ion ac o s egula e
conidia ion sugges ed a key ole in ansc ip ional egula ion
du ing conidia ion. In addi ion, se e al genes ha e been ound
o be highly exp essed du ing conidia ion (Robe s e al. 1988)
al hough hei mu an s do no ha e a clea de elopmen al
pheno ype. The con genes o N. c assa a e p e e en ially
exp essed du ing conidia ion bu some o hem a e also in-
duced by ligh in ege a i e mycelia in a WC-dependen man-
ne (Co ochano e al. 1995; Lau e and Russo 1991;Madi
e al. 1994). The genes con-10 and con-6 con ain se e al eg-
ula o y elemen s in hei p omo e s ha a e esponsible o
hei ansc ip ional egula ion by de elopmen , ligh and he
ci cadian clock (Co ochano e al. 1995; Lee and Ebbole
1998;Olmedoe al.2010b). Simila complex egula ion has
been obse ed o genes con-5 and con-13, since hey a e
egula ed by ligh and conidia ion al hough hei p omo e s
ha e no been ex ensi ely cha ac e ized.
The a ailabili y o he N. c assa genome sequence made
possible he c ea ion o a gene knockou mu an collec ion o
almos all he genes iden i ied in he genome (Colo e al.
2006; Dunlap e al. 2007). The iable knockou mu an s ha e
been analysed and hei de elopmen al pheno ypes ha e been
cha ac e ized (Bo ko ich e al. 2004; Ca illo e al. 2017;
Colo e al. 2006). Se e al s ains bea ing mu a ions in genes
coding o ansc ip ion ac o s ha e shown de ec s in
di e en s ages o he N. c assa li e cycle. Mos o hem we e
ound o ha e impai ed asexual spo ula ion al hough he co -
ela ion be ween he pheno ypes and gene exp ession du ing
conidia ion has no been s ablished o mos genes ye
(Ca illo e al. 2017).
In a ew cases, he ole o ansc ip ional egula o s on
conidia ion has been in es iga ed. Mu a ions in he Gal4-
like ansc ip ion ac o VAD-5 o he el e p o ein VE-
1 educe he g ow h o ae ial hyphae (Bay am e al. 2008;
Sun e al. 2012). The gene chc-1 encodes a p o ein wi h a
helix-loop-helix binding domain and he mu an shows
enhanced conidia ion, in pa icula unde high CO
2
(Sun e al. 2011).
Signal ansduc ion pa hways ha egula e
conidia ion
The key ole o he cAMP phosphodies e ase ACON-2 in
conidia ion indica ed ha o he signal ansduc ion pa hways
pa icipa ed in he egula ion o conidia ion in addi ion o he
ansc ip ional egula ion o conidia ion genes.
Conidia ion in N. c assa is also egula ed by signalling
pa hways based on he e o ime ic G p o eins and G p o ein-
coupled ecep o s (GPCR). G p o eins play key oles as sig-
nalling p o eins in euka yo es whe e hey o m he e o ime s
composed o α,βand γsubuni s which a e associa ed wi h
he plasma memb ane (Ne es e al. 2002; Won e al. 2012).
The Gαsubuni binds GTP and GDP and hyd olyses GTP o
GDP, and he Gβand Gγsubuni s o m a dime . In he inac-
i e s a e, he Gαsubuni binds GDP and he h ee subuni s
a e p esen in a complex in associa ion wi h a GPCR. Ligand
binding o he GPCR leads o exchange o GTP o GDP on
he Gαp o ein and dissocia ion o he Gαand Gβγ dime .
Bo h he Gα-GTP and Gβγ moie ies egula e downs eam
e ec o p o eins in a ious sys ems, including ion channels,
adenylyl-cyclases, phosphodies e ases and phospholipases.
GTP hyd olysis on he Gαsubuni allows he GDP-bound
Gα o eassocia e wi h he Gβγ dime and he GPCR a he
memb ane, eady o eini ia e he signalling cycle (Li e al.
2007).
Neu ospo a has h ee Gαsubuni s (GNA 1–3), one Gβ
p o ein (GNB-1), one Gγp o ein (GNG-1) and se e al p e-
dic ed GPCRs (Li e al. 2007). GNA-1 was he i s iden i ied
G p o ein subuni in ilamen ous ungi, and pa icipa es o-
ge he wi h he GPCR GPR-4 and he p o ein kinase A
(PKA) in he egula ion o ca bon sou ce-dependen apical
g ow h and asexual de elopmen (I ey e al. 1996;I ey
e al. 2002;I eye al.1999; Li and Bo ko ich 2006; Tu ne
and Bo ko ich 1993). GNA-3 egula es conidia ion ia mod-
ula ing a cAMP-dependen pa hway and ac s as a nega i e
egula o o conidia ion (Kays and Bo ko ich 2004; Kays
e al. 2000). The GNB-1 (Gβ)/GNG-1(Gγ) dime is equi ed
o he s abili y o Gαp o eins and ac s as a ep esso o
In Mic obiol
conidia ion (K ys o o a and Bo ko ich 2005;Yange al.
2002). Recen s udies ha e demons a ed ha RIC-8 ( esis an
o inhibi o s o cholines e ase) posi i ely egula es Gαsub-
uni s, GNA-1 and GNA-3, and dele ion o ic-8 esul s in
de ec s in g ow h and asexual de elopmen . Mu a ions ha
ac i a e he GNA-1 and GNA-3 p o eins in he Δ ic-8 back-
g ound pa ially supp ess Δ ic-8 pheno ypes. RIC-8 in e ac s
wi h GNA-1 and GNA-3 in he yeas wo-hyb id assays and
ac s as a GEF (guanine nucleo ide exchange ac o ) o GNA-
1 and GNA-3in i o (W igh e al. 2011). The pheno ypes o
mu an s in 36 GPCRs ha e been analysed. Mu a ions in 14
GPCR genes lead o al e a ions in conidia ion, speci ically in
he o ma ion o ae ial hyphae. Abou hal o he mu an s
showed a educ ion, and hal o he mu an had an inc ease
in he amoun o ae ial hyphae. Se e al mu an s had pleio o-
pic e ec s on de elopmen sugges ing c oss egula ions be-
ween hyphal g ow h, sexual and asexual de elopmen in
N. c assa (Cab e a e al. 2015).
Mu an s in componen s o o he signal ansduc ion pa h-
ways show al e a ions in conidia ion. Mu an s in genes coding
o calcium signalling componen s sugges a ole o calcium
signalling in conidia ion (Ba man and Tamuli 2017). The
GTP-binding cy oskele al p o eins sep ins pa icipa e in cell
pola i y and he s ains wi h dele ions in sep in genes show
al e a ions in conidia ion, among o he de elopmen al pheno-
ypes (Be epiki and Read 2013).
In addi ion, kinases pa icipa e in he egula ion o
conidia ion. The NDR kinase COT-1 egula es hyphal
b anching and in e ac s wi h se e al p o eins ha modula e
i s egula o y ac i i ies. The in e ac ing p o eins include
MOB2A/B, he a ginine me hyl ans e ase SKB1 and he eg-
ula o y subuni s o he p o ein phospha ase 2A. Mu a ions in
hese p o eins lead o changes in he egula ion o COT-1
ac i i y and al e a ions in conidia ion, sugges ing ha hey
play a egula o y ole in his de elopmen al p ocess (D ash
e al. 2010; Feldman e al. 2013; Shomin-Le i and Ya den
2017; Zi e al. 2013). The mu an in he second NDR kinase,
DBF-2, shows al e a ions in he g ow h o ae ial hyphae and
conidia ion (D ash e al. 2010), and a simila pheno ype is
obse ed in he mu an in he pu a i e his idine kinase DCC-
1 (Ba ba-Os ia e al. 2011).
Compa ison o conidia ion in N. c assa
wi h he asexual p og am o o he ascomyce es
The widesp ead occu ence o conidia ion in ascomyce es
highligh s he impo ance o his p ocess du ing he e olu ion
o his g oup o ungi (Be bee and Taylo 2001). N. c assa and
A. nidulans di e ged mo e han 300 million yea s ago (Taylo
and Be bee 2006; Taylo and Ellison 2010). Conidia ion in
A. nidulans has been in es iga ed in de ail and se e al mu an s
a ec ed on conidia ion ha e been cha ac e ized (Pa k and Yu
2012). These mu an s p o ide he ools o add ess ques ions
abou he e olu ion o conidia ion. Mo phologically, asexual
de elopmen in A. nidulans is signi ican ly di e en om ha
in N. c assa.InA. nidulans, conidia ion begins wi h he o -
ma ion o a oo cell and is ollowed by successi e eme gence
o he s alk, esicle, me ulae, phialides and inally ae ial
spo es o conidia (Pa k and Yu 2012). The mas e egula o
o conidia ion in A. nidulans is encoded by he gen b lA
(b is le). Al hough i has no sequence simila i y wi h l, he
ac i a ion o b lA ansc ip ion is su icien o induce
conidia ion, and in a simila manne o l,b lA is ac i a ed
by ligh in a WC-dependen manne (Mooney and Yage
1990; Ruge -He e os e al. 2011). B lA ac i a es ansc ip ion
o ano he egula o y gene, abaA (abacus). Unlike b lA,
o e exp ession o abaA does no induce conidia ion bu
does cause an a es o ege a i e g ow h and induc ion o
genes no mally obse ed du ing conidia ion (Adams e al.
1990;Adamse al.1998). The e a e many o he genes ac -
ing ups eam o B lA and whose mu a ion gi es ise o
aconidial pheno ypes. Mos o hese genes a e conse ed
in he Aspe gilli (Ojeda-Lopez e al. 2018). Some o hese
genes a e also ound in he N. c assa genome (Fig. 2)and
ha e been ound o complemen he de ec on spo ula ion
on he co esponding mu an s in A. nidulans (Table 1)
(Chung e al. 2011;Shene al.1998). Despi e he p esence
o se e al homologous genes be ween A. nidulans and
N. c assa, hei conidia ion pa hways di e in bo h hei
egula ion and mo phology, highligh ing he simplici y o
he conidia ion p og am in N. c assa (Fig. 2).
Conclusion
Conidia ion in N. c assa has a complex egula ion bu is a
simple de elopmen al p ocess when compa ed wi h ege-
a i e ep oduc ion in o he ilamen ous ungi. The isola-
ion o aconidial mu an s and he iden i ica ion o he genes
in ol ed showed ha di e en ial ansc ip ion and signal
ansduc ion pa hways ha e key oles in conidia ion. The
ole o en i onmen al egula ion in conidia ion is less un-
de s ood. The ole o ligh on he egula ion o conidia ion
by con olling ansc ip ion o l ando he key egula o y
genes has been p oposed, bu he connec ion be ween nu-
ien sensing and conidia ion, and how he ansi ion om
a liquid o an ai in e phase ac i a es conidia ion emain o
be in es iga ed in de ail. I is possible ha some o hese
en i onmen al signals a e sensed by any o he GPCRs ha
ha e been desc ibed in N. c assa.TheN. c assa genome
sequence and he collec ion o single mu an s wi h dele-
ions in mos o he genes in he genome ha e p o ided a
la ge collec ion o mu an s wi h al e ed conidia ion. The
la ge numbe o conidia ion mu an s sugges s ha al e ing
conidia ion is no e y di icul , bu iden i ying key egula-
o s, like FL o ACON-2, equi ed gene ic sc eens and
In Mic obiol

s ong mu an pheno ypes, no a educ ion o inc ease in
conidia ion. Fu u e wo k should ocus on iden i ying new
key egula o s o conidia ion pe haps by looking o condi-
ional aconidial mu an s.
The compa ison o he conidia ion pa hways in N. c assa
and A. nidulans shows a ew common ea u es like he egu-
la ion by ligh o key ansc ip ional egula o s, and he key
ole o ansc ip ional egula ion, bu many di e ences like
hose in mo phology and he use o speci ic p o eins in each
pa hway. I seems ha he e is no a basic conidia ion pa hway
sha ed by all ascomyce es bu di e en de elopmen al solu-
ions based on di e en ial ansc ip ion and common signal
ansduc ion pa hways o he p oblem o how o de elop co-
nidia. Howe e , we expec ha u he cha ac e iza ion o
conidia ion in N. c assa and compa ison wi h o he ilamen-
ous ungi will help o iden i y common elemen s in he eg-
ula ion o ungal de elopmen .
Acknowledgemen s Resea ch in he labo a o y o LMC is suppo ed by
he Spanish Minis y o Science, Inno a ion and Uni e si ies (BIO2015-
67148-R) and Eu opean Funds (Eu opean Regional De elopmen Fund,
ERDF).
Compliance wi h e hical s anda ds
Con lic o in e es The au ho s decla e ha hey ha e no con lic o
in e es .
Re e ences
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Table 1 Homologous genes be ween A. nidulans and N. c assa and he
co esponding pheno ype o he knockou mu an s
Aspe gillus
nidulans
Neu ospo a c assa Pheno ype o he N.c assa
mu an
s gA ––
luG NCU04264 Wild ype
lbA NCU08319 Wild ype
lbB NCU07379, c -5 Wild ype
lbC NCU03043, lb-3 Aconidial
lbD NCU01312, ca-1 Wild ype
lbE NCU05255 Unknown
adA NCU06493, gna-1 Wild ype
pkaA NCU06240, pkac-1 Pleio opic
s uA NCU01414, asm-1 Sho ae ial hyphae
osA NCU05964, os-1 Reduced conidia ion
medA NCU07617,
acon-3
Aconidial
b lA ––
abaA ––
we A NCU01033 Wild ype
–NCU8726, l Aconidial
–NCU9739, ld Aconidial
AN1251 NCU2713, csp-1 Conidial sepa a ion de ec i e
AN4878 NCU06095, csp-2 Conidial sepa a ion de ec i e
pdeB NCU00478,
acon-2
Aconidial
Fig. 2 Compa ison be ween he egula o y genes con olling conidia ion
in A. nidulans (in g ey) s. N. c assa (in blue). Conidia ion in A. nidulans
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conidia ion in A. nidulans a e also p esen in N. c assa genome al hough
he mo phological and egula o y p ocesses in bo h ungi subs an ially
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