Media and growth conditions

Chemicals for media preparation were dissolved in dH₂O and sterilized by autoclavation at 121°C for 20 min at two bar. Thermally unstable supplementations were dissolved in dH₂O and sterile filtered.

2.2.1 Bacterial growth

Escherichia coli strains DH5α^TM (Hanahan, 1985), DH10B^TM (Lorow and Jessee, 1990) and One Shot^® TOP10 (INVITROGEN) were used for construction and amplification of recombinant plasmids. Genotypes of these strains are given in TABLE 1. E. coli strains were cultivated in lysogeny broth (LB) (Bertani, 1951) medium (1% (w/v) tryptone, 0.5% (w/v) yeast extract, 1% (w/v) NaCl) on a rotary shaker at 37°C. Ampicillin was used as selective agent in a concentration of 100 mg/ml. Solid medium was prepared by supplementation with 2% (w/v) agar.

TABLE 1: E. coli strains used in this study.

Strain name Genotype

DH5α^TM F-, ∆(argF-lac)169, φ80dlacZΔM15, ∆phoA8, glnX44(AS), λ-, deoR481, rfbC1, gyrA96(NalR), recA1, endA1, thiE1, sdR17

DH10B^TM F- mcrA Δ(mrr-hsdRMS-mcrBC), φ80dlacZΔM15, ΔlacX74, deoR, recA1, araD139, Δ(ara, leu)7697, galU, galK, rpsL, endA1, nupG

One Shot^® TOP10 F- mcrA, Δ( mrr-hsdRMS-mcrBC), φ80dlacZΔM15, ΔlacX74, recA1, araD139, Δ(araleu)7697, galU, galK, rpsL (StrR), ndA1 nupG

2.2.2 Fungal growth

The A. nidulans veA⁺ strain AGB 551 (Bayram et al., 2012) was used as wildtype (WT) host for A. nidulans strains constructed in this study. A. fumigatus strain AfS35 (Krappmann et al., 2006b) was used as host for A. fumigatus strains constructed in this study.

A. nidulans and A. fumigatus strains were grown in minimal medium (MM) (1% (w/v) asexual development. Sexual development was induced by growing fungal strains in the dark for up to 7 d on solid MM plates, which were tightly sealed with Parafilm^® M (MERCK) to prevent oxygen supply.

A. fumigatus strains were grown in the presence of oxygen but without illumination. Strains were grown for 24 h in submerged cultures for synchronized growth. After this, mycelia were harvested and washed through sterile Calbiochem Miracloth filters (MERCK) and subsequently shifted onto solid MM plates (Fischer, 2002). Tests including the ztfA OE were carried out on MM supplemented with 10 µg/ml doxycycline to induce ztfA expression (Helmschrott et al., 2013). communication. A. nidulans strains are denominated AGB or FGSC, A. fumigatus strains are denominated ACS, AfS or AfGB.

Strain name Genotype Reference

FGSC A4 veA⁺ McCluskey et al., 2010

AGB551 ∆nkuA::argB, pyrG89, pyroA4, veA⁺ Bayram et al., 2012

AGB596 ^PgpdA::sgfp-phleo^R; pabaA1, yA2, veA⁺ Bayram et al., 2012 AGB1007 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆ztfA::six This study

AGB1008 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, six::^PniaD::ztfA This study AGB1009 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ztfA::sgfp::six This study AGB1010 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, sgfp::ztfA::six This study AGB1011 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ztfA::six This study

AGB1012 AGB1009, transformed with pME3173 This study

AGB1013 AGB1010, transformed with pME3173 This study

AGB1014 AGB551, transformed with pME3173 This study

AGB1015 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, This study

PztfA::ztfAS327A,T464A,S504-506A

::six

AGB1016 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆fluG::six This study AGB1017 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆fluG::six, ∆ztfA::six This study AGB1018 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆fluG::six,

six::^PniaD::ztfA

This study AGB1019 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, six::^PniiA::fluG This study AGB1020 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆ztfA::six,

six::^PniiA::fluG

This study AGB1022 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆aflR::six This study AGB1023 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆aflR::six, ∆ztfA::six This study AGB1024 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆aflR::six,

six::^PniaD::ztfA

This study AGB1025 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, six::^PniiA::aflR This study AGB1026 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆ztfA::six,

six::^PniiA::aflR

This study AGB1028 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆abaA::six This study AGB1029 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆abaA::six, ∆ztfA::six This study AGB1031 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆brlA::six This study AGB1032 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆brlA::six, ∆ztfA::six This study AGB1033 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆brlA::six,

six::^PniaD::ztfA

This study AGB1035 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbB::six This study AGB1036 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbB::six, ∆ztfA::six This study AGB1037 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbB::six,

six::^PniaD::ztfA

This study AGB1039 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbC::six This study AGB1040 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbC::six, ∆ztfA::six This study AGB1041 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbC::six,

six::^PniaD::ztfA

This study AGB1043 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbD::six This study AGB1044 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbD::six, ∆ztfA::six This study AGB1045 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbD::six,

six::^PniaD::ztfA

This study AGB1047 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbE::six This study AGB1048 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbE::six, ∆ztfA::six This study AGB1049 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆flbE::six,

six::^PniaD::ztfA

This study AGB1051 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, phleo^R,

PniaD::ztfA::eyfp-C, ^PniiA::rcoA::eyfp-N

This study

AGB1052 AGB1051, transformed with pME3173 This study

AGB1053 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, phleo^R,

PniaD::ztfA::eyfp-C, ^PniiA::eyfp-N

This study AGB1054 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, phleo^R, ^PniaD::eyfp-C,

PniiA::rcoA::eyfp-N

This study

AGB1055 AGB1053, transformed with pME3173 This study

AGB1056 AGB1054, transformed with pME3173 This study

AGB1057 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆vosA::six S. Thieme, p.c.

AGB1058 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆vosA::six, ∆ztfA::six This study AGB1059 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆vosA::six,

six::^PniaD::ztfA

This study AGB1062 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆velC::six This study AGB1063 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆velC::six, ∆ztfA::six This study AGB1064 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆velB::six S. Thieme, p.c.

AGB1065 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆velB::six, ∆ztfA::six This study AGB1066 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆veA::six J. Gerke, p.c.

AGB1067 ∆nkuA::argB, pyrG89, pyroA4, veA⁺, ∆veA::six, ∆ztfA::six This study

ACS39 ∆akuA::loxP, ΔpyroA::pyroA-TetOn-ztfA C. Sasse, p.c.

AfS35 ∆akuA::loxP Krappmann et al., 2006b

AfGB129 ∆akuA::loxP, ∆ztfA::six This study