Table 3.

List of A. fumigatus mutants tested in Galleria mellonella.

Function			Mutants	Reference A. fumigatus strain	In vitro effects on development, stress response, and metabolism	Virulence in G. mellonella	Virulence in mice	Ref.
	SECONDARY METABOLITES
mtfA gene encodes a putative C2H2 zinc finger domain-type transcription factor (fungal development and secondary metabolism)			∆mtfA	Af CEA10	Role in growth rate & gliotoxin production	Hypovirulent	/	[110]
			OEmtfA		Role in growth rate & conidiation, stress response, gliotoxin production	Normal	/
rtfA gene encodes a RNA polymerase II transcription elongation factor-like protein			∆rtfA	Af CEA17	Reduce growth rate, increase conidiation, oxidative stress, and metabolites metabolism	Hypovirulent	Normal (40)	[97]
			rtfA OE		No difference in growth, conidiation, stress response, or metabolites metabolism. Minor effect on cell wall.	Normal	/
Gene cluster involved in melanin pathway		Alb1: encodes polyketide synthase	Color mutant alb1	Af 293	/	Hypervirulent	Hypovirulent	[100]
		Ayg1: encodes heptaketide hydrolyase	Color mutant ayg1			Hypervirulent	/
		Arp2: encodes hydroxynaphthalene reductases	Color mutant arp2			Hypervirulent	/
		Arp1: encodes scytalone dehydratases	Color mutant arp1			Hypervirulent	Hypovirulent
		Abr1: encodes multicopper oxidase	Color mutant abr1			Hypervirulent	/
		Abr2: encodes laccase	Color mutant abr2			Hypervirulent	/
dmaW, easM, A, G genes involved in ergot alkaloid pathway			∆dmaW	Af FGSC A1141	/	Hypovirulent	/	[109]
			∆easM	Af293		Hypovirulent	/
			∆easA/G			Hypovirulent	/
Pes1, PesL: nonribosomal peptide (NRP) synthetases involved in fumigaclavine C biosynthesis			∆pesL	∆akuB mutant background	Reduce tolerance to H2O2 but increase tolerance to menadione, essential for fumigaclavine C biosynthesis	Hypovirulent	/	[107]
			∆pes1	in ∆akuB mutant and ATCC 46,645 backgrounds		Normal	/
			∆pes1	Af293.1	Alter conidial morphology & hydrophobicity. More susceptible to oxidative stress	Hypovirulent	/	[108]
	DEVELOPMENT
Rgs (RgsC, RgsD & GprK): regulator of G protein signaling, crucial roles in upstream regulation of vegetative growth, development, secondary metabolism, and virulence			∆rgsD	Af293	Increase conidiation, stress response, gliotoxin and melanin production	Hypervirulent	/	[87]
			∆rgsC	Af293.6	Increase germination, reduce conidiation, growth, tolerance to H2O2 and gliotoxin production, modify cell wall	Hypovirulent	/	[76]
			∆gprK	Af293.1	Increase germination & reduce conidiation, tolerance to H2O2 and gliotoxin production	Normal	/	[77]
NosA: number of sexual spores, transcription factor			∆nosA	Af CEA17 ∆akuB	Slightly reduce conidiation and increase radial growth & germination	Hypervirulent		(115)
Cofilin: actin depolymerizing factor, role in actin cytoskeleton dynamic			cofilinteton/cofilin S5E	Af CEA17 ∆ku80	Impair growth rate, regulate cell wall & modify resistance to H2O2	Hypovirulent	Normal	[86]
			Cofilin OE		Role in growth rate & cell wall, increase resistance to H2O2	Normal	Normal	[85]
			D19A R21A		Increase production of ROS, apoptosis and ergosterol levels	Hypervirulent	/	[89]
			K36A		Increase production of ROS, apoptosis and ergosterol levels	Hypervirulent	/
Myosin: cytoskeleton component, member of actin-based motor proteins family			∆myoB (class II)	Af akuBKU80 pyrG−	Delayed germination, increase of conidiation & modification of cell wall	Hypovirulent	Hypovirulent	[95]
			∆myoE (class V)		Delayed germination, reduction of growth rate & conidiation, modification of cell wall	Hypovirulent	Normal
			∆myoB∆myoE	∆myoB		/	/
Septins: GTPases family, regulates cellular processes (cell wall integrity, septation)			∆aspA	Af akuBKU80	No difference in growth rate, reduce conidiation	Hypervirulent	/	[94]
			∆aspB		Reduce growth rate, reduce conidiation	Hypervirulent	Normal
			∆aspC		No difference in growth rate, reduce conidiation	Hypervirulent	/
			∆aspD			Normal	/
			∆aspE			Normal	/
			∆aspE∆aspD			Normal	/
			∆aspE∆aspB			Hypervirulent	/
			∆aspE∆aspD∆aspB			Hypervirulent	/
Rho1: small GTPase, a potential regulatory subunit of β-1,3-glucan synthase			Rho1teton	Af CEA17Δku80	No growth & increase H2O2	Avirulent	/	[96]
			Rho1 OE		No difference in growth rate or in stress response	Normal	Normal
Kin1: protein kinase is a member of the eukaryotic PAR-1/MARK/MELK family			∆kin1	Af akuBKU80	No difference in growth rate, conidiation & cell wall. Role in stress response	Normal	/	[91]
srgA: (secretion related GTPase A) encoding a Rab GTPase of the Rab family, master regulators of membrane trafficking			∆srgA A	AfS28	Reduce growth rate & aberrant conidiation, role in stress response	Normal	/	[97]
			∆srgA B			Normal	/
			∆srgA C			Hypovirulent	/
ERMES: Endoplasmic-Reticulum mitochondria encounter structure, tether between mitochondria and endoplasmic reticulum			mmm1tetOn	AfS35	Reduce growth rate	Hypovirulent	/	[98]
Calcineurine		swoH: nucleoside iphosphate kinase, interaction with the catalytic subunit of calcineurin (CalA)	SwoHV83F	Af CEA17-80	Reduce growth rate and increased sensitivity to elevated temperatures	Hypovirulent	/	[85]
		cnaA: calcineurin A (cnaA) catalytic subunit	∆cnaA	Af293.1	Reduce germination, growth rate & conidiation	Hypovirulent	Hypovirulence	[88]
	METABOLISM
mirC: Siderophore iron transporter involved in synthesis of siderophores in intracellular microsomal compartment			∆mirC (Iron-deplete condition)	Af ATCC 46,645	Reduce growth rate and conidiation	Normal	/	[66]
Sid: siderophore CpcA: cross pathway control (transcriptional acgtivator) Paba: gene encodes para aminobenzoic acid synthetase PABA, in folate biosynthesis pathway			∆sidA	Af ATCC 46,645	/	Avirulent	Avirulent	[58]
			∆sidC	Af ATCC 46,645		Hypovirulent	Hypovirulent
			∆sidF	Af ATCC 46,645		Avirulent	Hypovirulent
			∆sidD	Af ATCC 46,645		Hypovirulent	Avirulent
			cpcA	Af D141		Avirulent	Hypovirulent
			paba	Af 237		Avirulent	Avirulent
ArgEF: encodes for 3 enzymes acetylglutamate synthase & kinase, acetylglutamyl-phosphate reductase ArgB: encodes for ornithine transcarbamoyl transferase			∆argEF	∆akuB derivated from Af CEA17	Reduce growth rate & iron metabolism	Hypovirulent	/	[112]
			∆argB	Af293	No difference in growth & iron metabolism	Normal	/
ArgJ: the only arginine biosynthetic enzyme lacking mammalian homologs			∆argJ	AfS77 (ATCC46,645)	/	Hypovirulent	Hypovirulent	[113]
AmcA: mitochondrial transporter			∆amcA (Iron-deplete condition + Nitrogen source)	Af S77 (ATCC 46645 ∆KuA)	Reduce conidiation, growth rate & iron metabolism in presence of Glutamate or Ornithine	Normal	/	[60]
					No difference in fungal development & slight reduction of iron metabolism in presence of Arginine or Arginine + Ornithine		/
AcuM: zinc cluster transcription factor			∆acuM (Iron deficiency condition)	Af293	Reduced growth rate and iron metabolism	Hypovirulent	Hypovirulent	[65]
LeuB: transcription factor			∆leuB (Normal or Iron deficiency condition)	Af A1160 ΔKu80 pyrG	Reduce growth rate in normal condition	Hypovirulent	/	[59]
					Increase or reduce growth rate in iron deficiency condition		/
PptA: a phosphopantetheinyl transferase (P-pant)			∆pptA	Af A1160 ΔKu80 pyrG+	No production of secondary metabolites	Avirulent	Avirulent	[69]
PcaA: PIB-type cation ATPase which links metal homeostasis and heavy metal tolerance			∆pcaA	Af293	Reduce growth rate in presence of cadmium sulfate	Hypovirulent	/	[67]
					No difference in growth rate in presence of copper, iron, silver & zinc sulfate
			OEpcaA	Af293	Increase growth rate in presence of cadmium sulfate	Normal	/
					No difference in growth rate in presence of copper, iron, silver & zinc sulfate
Siroheme: heme-like group used for sulfate and nitrate assimilation; met8: gene encoding the bifunctional dehydrogenase/ferrochelatase enzyme Met8			∆met8	AfS77	Role in growth rate & stress response	Hypovirulent	/	[61]

The analysis of the effect of virulence factors is based on the comparison of virulence of a wild strain with that of mutants obtained by gene deletion.

Increased: increased pathogenicity relative to wild strain; Decreased: decreased pathogenicity relative to wild strain;

No change: no change in the pathogenicity relative to wild strain; Avirulent: no pathogenicity with the mutan