Posted on Authorea 3 Aug 2020 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159646770.00457953 | This a preprint and has not been peer reviewed. Data may be preliminary. n nteitrcin ihohrognss ciga ois hmclmsegr,srcua components structural messengers, chemical toxins, as of acting physiology organisms, the other in with roles interactions important the play in Terpenoids and 2019). Mukherjee, & Sherkane, terpenes 2016; (Pachauri, and Paikrao, polyketides peptaibols, 2008). & wide with a al., Patil, Trichoderma 2005), produce et Patil, Keller, fungi (Reino & 2014; relevant These roles (Yu most 2019). al., biological manner Sharma, the strain-dependent the et & as a and Rialch, Hermosa in Salwan, 2007), SMs 2018; 2019; Mac´ıas-Rodr´ıguez, al., of al., Surapathrudu, variety (Contreras-Cornejo, et del & et reviewed Viterbo Solanki, Contreras-Cornejo Solanki, extensively 2008; 2016; Rai, al., been Larsen, et has & Vinale metabolites Val, 1996; these al., protect Mastouri, to to et & able associated (Lorito Harman, also produce (Shoresh, are and they responses of 2016), defence (SMs) effects al., plant beneficial et the The phytopathogens (Fiorini inducing of growth by 2010). range plant indirectly wide promote pathogens a strains mycoparasitize against Some and plants antagonize 2004). to al., ability their et to (Harman due protection plant crop in Trichoderma INTRODUCTION different which in functions. picture biological a different Trichoderma provide of studies driver indication Expression the strong is Trichoderma. a is genes of ways, members genomes TS many family the of in gene in regulation regulated chemotypes of found the are terpenoid differentiation genes genes and of functional TS variety TSs TS family that of a on TS-gene number revealed propose we studies high enzymes We the clade-specific Here, the expression within demands. of for diversity gene ecological presence characterized. the different the and cover of and been spp., overview TSs, to Trichoderma not of evolved An 17 groups yet 15 model. have to identified Terpenoid a We biosynthesis belonging hosts, as valueprovided. their strains plant produced. T6085 and high 21 in they gamsii fungal involved of with T. their metabolites TSs genes with and using secondary of and (TS) protection environment the genomics synthase the plant by comparative with terpene crop combined supported spp. the Trichoderma are in of of species agents interaction most the these biocontrol however in of players as key effects used are beneficial compounds species The comprising industry. genus for fungal a is Trichoderma Abstract 2020 3, August 3 2 1 Sarrocco Puntoni Grazia Vicente Isabel inventory synthases Trichoderma terpene in the into insights provide biology approaches molecular and genomics comparative Combined nvriyo Salamanca of University Salamanca de Universidad Pisa of University 1 sagnso bqiosfni opiigbnfiiu pce sda icnrlaet (BCAs) agents biocontrol as used species beneficious comprising fungi, ubiquitous of genus a is p.aerpre opoueaboddvriyo epnis nldn oaiecompounds volatile including terpenoids, of diversity broad a produce to reported are spp. 1 icroBaroncelli Riccardo , 1 oaHermosa Rosa , Trichoderma 3 niu Monte Enrique , 2 a´aEgnaMor´an Mar´ıaDiez Eugenia , p.aespotdadotnrl ntescnaymetabolites secondary the on rely often and supported are spp. 1 3 ivniVannacci Giovanni , 3 oof Bernardi Rodolfo , 1 n Sabrina and , Trichoderma 1 , Posted on Authorea 3 Aug 2020 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159646770.00457953 | This a preprint and has not been peer reviewed. Data may be preliminary. nePocnv.47. Jnse l,21)wsue oietf Spoen in proteins TS identify to used was 2014) al., bassiana et (Jones v5.44-79.0 InterProScan proteins TS of Prediction 2. with relation 17 to Table Information Infor- belonging Biotechnology strains for 21 Center prising (National databases public in available mation, annotation, gene including - Genomes 2020 al., platform et Genomic (Sarrocco 1. responses defence with plant METHODS interaction the AND beneficial MATERIALS inducing a establishes and the fungus endophyte and the an al., symptoms addition, as unpublished). In et FHB behaving unpublished). reducing Sarrocco - roots, and 2019, 2020 wheat ppm) Battilani, al., pathogen (50 et & the concentrations Sarrocco Mauro, of 2013b, DON growth (Sarrocco, high the (DON) reduce of of deoxynivalenol to development presence able of in is production growing T6085 the wheat. 2013a), on as (FHB) well against Blight BCA as Head as Fusarium evaluated of been agent has it causal years, the using ten regulating past fungi, signals the these environmental During of the 17 ecology decipher the complete to to to the aimed belonging of addition, in characterization strains In genes genomic genus. TS 21 the the of on of potential focused genes biosynthetic we TS of family, 2019). TS-gene ecology of al., the the et set within Kubicek in diversity 2011; terpenoids the al., of about et Kubicek importance 2013; the al., Given et (Mukherjee quantitative merely been – has species three to limited in family TS-gene complete and 2013), in al., et sesquiterpenes (Pilsyk in synthase com- pyrophosphate trichodermin first farnesyl and the A experi- catalyses Other harzianum been which 2011a,b). trichothecenes have gene, of family (TRI5)-encoding biosynthesis brevicompactum TS synthase T. the the trichodiene in of the in step on members genes TS mitted focused few of mainly only characterization Functional been and 2016). has biosynthesis, Mukherjee, & their (Bansal from in characterized involved mentally exclusively isolated alkaloids). genes indole those been TS and in have and terpenoids about distinguished indole sesterterpenes terpenes be C25 (meroterpenoids, many can origin diterpenes, Although mixed C20 terpenes of sesquiterpenes, scaffolds, those C15 their and monoterpenes, (Guzm´an-Ch´avez of triterpenes), biosynthesis (C10 origin C30 units terpenoid the isoprenyl in to by steps trans- According formed committed prenyl and 2018). the (TCs) mediating al., cyclases enzymes terpene et (IPSs), by core synthases modifications the pyrophosphate further (PTs), undergo isoprenyl ferases that iso- the precursors & by pyrophosphate (C) the Flynn, synthesized geranyl constitute (Quin, carbons are and (GGPP) 10C molecules pyrophosphate five pyrophosphates: geranylgeranyl These polyprenyl the 20C 2014). terpenes and and linear Schmidt-Dannert, are (FPP) fungal of Isopentenyl-pyrophosphate pyrophosphate acetyl-coA, biosynthesis all farnesyl et (TSs). from 15C variety, the (Pachauri (GPP), enzymes for synthetized huge responses synthase blocks their both defence building terpene pyrophosphate, Despite prene plant by dimethyl-allyl 2016). of precursors isomer Mukherjee, inducers few its & and from Bansal, stress synthesized to Gruber, are related Zeilinger, genes 2019; of al., regulators membranes, of www.ncbi.nlm.nih.gov rtoe,bsdo h em soitdt hi osre oan Iorni ytaedomain: synthase (Isoprenoid domains conserved their to associated terms the on based proteomes, Trichoderma Trichoderma .graminearum F. 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S1 epciey(adz ta. 01 amec ta. 03 04 05 ieioe al., et Tijerino 2015; 2014, 2013, al., et Malmierca 2011; al., et (Cardoza respectively , .Mdlorganism Model ). easse h xrsinpten fsm S ndffrn odtosassociated conditions different in TSs some of patterns expression the assessed we , .virens T. spp. Trichoderma Cuce ta. 03.Frhroe eoemnn tde aeasse the assessed have studies mining genome Furthermore, 2013). al., et (Crutcher on eoeInstitute, Genome Joint ; Sgnseprmnal hrceie are characterized experimentally genes TS eihcao ha ta Mtrs ta. 00 02 arcoe al., et Sarrocco 2012; 2010, al., et (Matarese straw wheat on perithecia Trichoderma , .gamsii T. .atroviride T. euei bassiana Beauveria oee ntoecss h iest ftegnshsbe mainly been has genus the of diversity the cases, those in however , 68 samdl tanT05peet estl lifestyle. versatile a presents T6085 Strain model. a as T6085 Trichoderma pce,wr sdfrcmuainlaaye (Supporting analyses computational for used were species, and Trichoderma 2 .reesei T. Trichoderma www.mycocosm.jgi.doe.gov/mycocosm/home vir4 a sda ugopdeiscoephylogenetic close its due outgroup as used was p. rvdn noeve fteterpenoid the of overview an providing spp., uaimgraminearum Fusarium eurdfrteboytei fmn-and mono- of biosynthesis the for required , Bna uhre 06,o h study the or 2016), Mukherjee & (Bansal – pce,teei oetnieinformation extensive no is there species, n h creifrainavailable information scarce the and , erg-20 Trichoderma of .reesei T. .arundinaceum T. h otaggressive most the , Trichoderma noiga encoding , p.and spp. ,com- ), and B. Posted on Authorea 3 Aug 2020 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.159646770.00457953 | This a preprint and has not been peer reviewed. Data may be preliminary. i)Sbtaeseict n uaiefntoswr sindb hlgntcanalysis: phylogenetic by assigned sequences; amino-acidic were the functions in bifunctional (pHMM) putative and and Models mono- and Markov of Hidden Substrate-specificity 2012) diffe- profile Peters, iii) enabled & for Class Honzatko, turn searching to (Gao, identified in TS-folds associated were which (DxDD) motifs enzymes motifs, II metal-binding Panther Class aspartate-rich and and Conserved (D[D/E]xx[D/E]) ii) Pfam I proteins; on TCs based of identified identification were rentially proteins PTs in i) enzymes approaches: TS characterize to order proteins. proteins In TS TS to membership, of associated superfamily characterization regions sites, (TM) Genomic to Panther trans-membrane 3. relatives and and motifs domains, identify Prosite to transferase PIRSF, used prenyl were of Pfam, interface, variants IPR008930). the in domain: included transferase algorithms cyclase/prenyl Terpene IPR008949, fseie00%Ten8 ouin n 10 biological a Three 16 i.e. and photoperiod weeks). solution, condition, and with Tween-80 (5 each chamber 0.01% stage per growth sterile anthesis a of inoculated the in were reached incubated plants each and until plants + respectively, mix three 20/22 potting of at a replicates dark, in h pots light/8 days. in h 4 sown condition. for were each seeds rpm for Wheat 180 included at were shaker replicates N MM rotatory biological liquid and independent in a 6. frozen Three sucrose, on Millipore), 0.9% extraction. 28 (475855-1R, RNA MM at Miracloth until MM, incubated using -80 of were filtration at H by mL Flasks mM discarded. collected 25 wash was 0.5 NaCl. were containing to Mycelium as mM min supernatants flasks such 200 10 mL and for stressors, and 50 min rpm starvation), different in 10000 10 adding at inoculated for by centrifugated then and rpm modified was water 10000 distilled Mycelium at sterile in it. centrifugation resuspended by was collected pellet (MM) Mycelial was medium Mycelium minimal of h. mL 10 25 of containing concentration flasks L mL L g 50 g 0.13 in (9 inoculated sucrose and 0.9% plates PDA 1-week-old from at water of distilled Mycelium sterile in of by stored cultures followed were shaking, Liquid Seeds on water. 5. min vernalization. distilled 3 for sterile for days with wheat chlorine) 3 each experiments, active min for all (0.6% 10 4 Before solution of 20/22. NaClO steps at a washing dark, with three light/8h sterilized h 16 surface photoperiod were with seeds chamber growth a in incubated from Seeds light/darkness. nearum gamsii Trichoderma material plant InterProScan and using Fungal domains 4. conserved for model screened evolutionary trees above. individually corresponding Phylogenetic were I described and 50%. alignments identified as + for were cluster WAG used threshold phylogenetic Sequences values the each (Abascal, evaluated. conservation to ProtTest bootstrap using topology and using and and reconstructed checked probabilities obtained 2010) visually was The was were Arkin, 2001). tree model Goldman & substitution Phylogenetic & Dehal, best 2005). (Whelan (Price, The Posada, v2.1.11 2010). & FasTree al., Zardoya, 2003), et Table Huelsenbeck, (Guindon Information & v3.3.2 (Supporting PhyML (Ronquist literature MrBayes in with: described built and functions known of Fgra Tgam .bassiana B. Tgam hss For theses. -1 TM14( 124 ITEM neatosi H scenario FHB in interactions CaCl + Tgam Fgra 2 Spoen eeaindb AF 740(ao ta. 03 ln ihT proteins TS with along 2013) al., et (Katoh v7.450 MAFFT by aligned were proteins TS . L g 0.1 , lns lnswr oee ihawiebg rvosymitndisd ihwater with inside moistened previously bag, white a with covered were Plants plants. Tgam a bandfloigatose iudcluepoeue prsof Spores procedure. culture liquid two-step a following obtained was 6 -1 Fgra 68 ( T6085 prsml spores urs,1gLNH g/L 1 sucrose, Tgam nclto,soe eecletdb ahn -ekodPApae ih2 mL 20 with plates PDA 1-week-old washing by collected were spores inoculation, -1 Zpaaae l,21)wr rw nPA(im)pae t2,1 /2h h/12 12 25, at plates (Sigma) PDA on grown were 2017) al., et (Zapparata ) al .13gL g 0.0183 NaCl, rtcmaestivum Triticum Tgam -1 lsswr nuae t2 narttr hkra 8 p o 60 for rpm 180 at shaker rotatory a on 28 at incubated were Flasks . Brnel ta. 06 aaeee l,21)and 2012) al., et Matarese 2016; al., et (Baroncelli ) Trichoderma 4 NO 7 -1 prsmL spores cv. 3 FeSO L g 5 , Apogee p.and spp. 3 4 .05gL g 0.0035 , -1 -1 C 2 4 wet eesw npt naptigmxand mix potting a in pots in sown were (wheat) upninwssrydo pksof spikes on sprayed was suspension O H 2 .bassiana B. 12 ny00%o NH of 0.01% only , N 2 O -1 6 L g 1 , ZnSO Tgam eue obnto fdifferent of combination a used we 4 .0 L g 0.002 , -1 alone, K S2 2 HPO 4 NO .Pyoeei rewas tree Phylogenetic ). Fgra 3 4 -1 Tgam . L g 0.5 , n C and Trichoderma MnCl uaimgrami- Fusarium ln and alone eecollected were 4 2 2 Tgam H tafinal a at ) -1 n stored and 12 N MgSO 2 Tgam O alone spp. 6 (N 4 , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. .Caatrzto fTspoie noeve ftetreodboytei oeta in of potential genomes biosynthetic the terpenoid in found the an of used overview We an provides TSs of Trichoderma Characterization 1. RESULTS of patterns formation. ANOVA expression dimmer for and assessing used for and used consistent Primers ts9 were test. replicates Tukey biological and SYSTAT ANOVA three using from Values analysis, statistical 2001). Schmittgen, as such & genes, (Livak housekeeping in the other among performed using stability were expression calculated reactions were 95 PCR (Ct) denaturation, (Qiagen) All cycles of cycler 20 cycles (Qiagen). Q of 40 2X volume Rotor-Gene min; Mix total in a Master performed in PCR PCR triplicate Green real-time SYBER(r) quantitative QuantiNova by with analyzed Maxima using was synthesis expression instructions. cDNA manufacturer’s Gene for the analyses used to were expression according RNA Gene biosystems) of Applied 9. according ng (K1642 removal, 400 kit gDNA synthesis of for cDNA total Sigma-Aldrich) Strand A AMPD1 First Grade, were instructions. samples Amplification manufacturer’s RNA I the RNeasy (1987). (DNase to Willmitzer I and the Schell DNase Logemann, using with by in treated described extraction ground method were the RNA fungus to the according total by extraction colonized for spikes Wheat N used N instructions. liquid liquid were manufacturer’s the in powder to ground according of (Qiagen), were mg by plates 100 colonized control containing roots interaction wheat root cultures, - liquid from biomass Fungal manipulation same acid the Nucleic condition. in each days 8. Plates for 3 included for from were dish. incubated Mycelium replicates Petri and biological extraction. the tape Three by RNA cm with of colonized until 1 sealed roots -80 centre h, were Wheat at 24 the Plates above. After from controls. described as distance 22. conditions plants at of without h cm plates 24 PDA 1.5 for at of incubated colony and PDA 1-week-old tape on with placed sealed were were seeds wheat (P) Four value (P transformation, angular healthy after ANOVA of 7. of by percentage inoculation in statistically the after determined DS), calculating days were – by Six values evaluated severity DS was for (Disease h. symptoms removed spikelets 24 FHB then symptomatic additional of were and for Reduction Bags back extraction. h. placed RNA in 24 were until incubated additional bags were for white Plants above 1h, pathogen. Fgra described after the conditions by and same penetration aeration facilitate the plant to in above described chamber placed growth conditions was a bag same 10 black the a a and and in water, solution, h Tween-80 personal 0.01% 48 M., of sterile (Dufresne for Fgra conidia of fungus chamber pathogen, mL the growth the 20 by a of with penetration inoculation in facilitate For incubated to above. bag were black Plants a with communication). and humidity, maintain to Tgam , ln and alone ts11 e pksclnzdb ug eecletdfo ahcniin rzni iudN liquid in frozen condition, each from collected were fungi by colonized spikes ten , 2 sn r-hle otradpsl.Smlscnann 0 go odrwr sdfrttlRNA total for used were powder of mg 300 containing Samples pestle. and mortar pre-chilled using , ha ot interaction roots wheat – tri5 in-silico Tgam ee n hs of those and genes Tgam prahi re oasg uaiefntost 8 Secddpoen currently proteins TS-encoded 387 to functions putative assign to order in approach + Fgra Trichoderma a lcdi h eteo h ltscnann ha edig,a ela in as well as seedlings, wheat containing plates the of centre the in placed was lns lnswr oee ihawiebg rvosymitndisd with inside moistened previously bag, white a with covered were Plants plants. μ ° o 0cce ne h olwn odtos nta ciain 95 activation, initial conditions: following the under cycles 40 for L β- o e n obndanaigetnin 60 annealing/extension, combined and sec 5 for C © tubulin .32sfwr.Dt rmlqi utrsof cultures liquid from Data software. v.13.2 p.Iiily eietfidP n Cpoen codn otheir to according proteins TC and PT identified we Initially, spp. β- tubulin Spotn nomto Table Information (Supporting Tgam eea noeoscnrl hc a eetddet its to due selected was which control, endogenous as gene Fgra D oto ltswscletdadsoe ni use. until stored and collected was plates control PDA 4 Fgra Tgam 5 eecletdb ahn -ekodPAplates PDA 2-week-old washing by collected were 2 prsmL spores ln and alone sn r-hle otradpsl.Samples pestle. and mortar pre-chilled using actin eecletd rzni iudN liquid in frozen collected, were Tgam and -1 n uglmclu rmthe from mycelium fungal and Fgra tef-1 2 upninwssrydo pksof spikes on sprayed was suspension grpu u u ttebre of border the at out cut plug agar S3 ts1 + aawr xrse s2 as expressed were Data . Tgam eecekdfrefficiency for checked were ) , ts3 Tgam lns ieecson Differences plants. , ° ,1 e.Threshold sec. 10 C, ts4 (r) 2 n trda -80 at stored and a nlsdby analysed was , ts5 ln iiKit Mini Plant 2 < , n stored and ts6 0.05). , Tgam -ΔΔ῝τ ° ts7 ,2 C, , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. 01.Frhroe hs Sscnan5cnevdQ ois hc r huh ob epnil of responsible be 2000). to al., et thought Noguchi, (Kushiro are & intermediates Takagi, which carbocation the Naito, motifs, stabilize (Abe, QW and enzyme enzymes conserved the these 5 of in contain structure the reported OSCs strengthening aspartate-rich DCTAE these DCISE classical Furthermore, or DCTSE the showing 2001). of 1) variants Fig. both in in colour motifs, present (light-brown Table PS01074) is et Information PTHR11764; SQS (Linscott (Supporting 13243; additional Pfam pathway-specific reticulum in an probably endoplasmic of conserved is TSs, the genomes universally them these to All is of of protein which one one the least residues, contains at binding 23 genome that of of each indicating responsible region Although is helix (red 2016). it TM al., 2018) and C-terminal al., SQSs a geranyl- et eukaryotic identified II PTHR11626:SF2; (Jeong all also type (PTHR11774:SF6). (FTases) (PTHR11774:SF4), prenylation we 00494; transferases 1) protein (GGTases SQSs, farnesyl (Pfam transferases and in In geranylgeranyl (SQSs) (PTHR11774:SF11) involved I 2) synthases enzymes type (GGTases as transferases squalene and such geranyl as 1) 1), this identified Fig. by Fig. in PTs, colour in composed of colour also clade, group (orange is Harzianum large PS01044) group the this a to suggests found belonging sesquiTS We species of of groups exclusive TSs. both widely be of to are type to proximity 4” phylogenetic seems their group 3” in and “uncharacterized group represented “uncharacterized could of particularly they of TSs are suggests but proteins This species, pentalenene. motif. the DDxxD than motifs, across additional metal-binding others distributed conserved an contain highly sesquiterpenes but share synthesize triad lasts these NSD/DTE actually of the most on lack Although proteins analysed. some genomes the (“unchar- all Table proteins in species, known found Information monoterpene Viride with (Supporting and cluster in not respectively) absent sesquiTS did are “4”, in that and found proteins of 3” be groups acterized presilphiperfolan-8 two can 16 with which including sharing along sesquiTS proteins 03936), synthases, of I pentalenene (Pfam groups Class two contains domain contains 1) It C (monoTS). Fig. while synthases synthase in 2”, colour terpene (light-green group conserved group “uncharacterized TRI5-superfamily a the of of of TSs clade species two sister the some The found and from we Viride species Viride, in among present of only conserved species were In arundinaceum highly 1” found group clade. “uncharacterized were Longibrachiatum of which the proteins synthases, Instead, as longiborneol clades. clade, named other lacking Brevicompactum therefore only the were the of and were species TSs, in in known found trait with was monophyletic cluster TRI5 not respectively. gamsii did “2”, T. that (Pfam (Support- and clade proteins 1” superfamily Viride group of TRI5 of “uncharacterized groups species the two in to represented and particularly belonging synthases, was (sesquiTSs) Table which synthases 1), Information Fig. sesquiterpene ing in of enzymes. colour TSs, bifunctional group (dark-green known 06330) are vast Table with Information they a (Supporting indicates cluster found Viride motifs We not clade DxDTT to did belonging II they species Class Although of and exclusive S4 DDxxE be 1). to I seems Class these Fig. Interestingly, both in C-terminal of and colour presence PTHR43611:SF3) (light-blue the 13419; (Pfam (IPR008930) HAD-like domains N-terminal TS sharing are TSs numbers specific accession revealed phylogenetic protein Analysis including same tree Table the phylogenetic Information and in inventory, Supporting clustered TS in the available motifs of metal-binding (Fig. portions colour and specific different showing groups domains in 15 highlighted conserved to one characterized functions sharing each putative biologically group, TSs assigning using as well analysis proteins. as phylogenetic Class substrate-specificity, of Clustering-based determining as enabled classification enzymes. also protein TSs Bifunctional enabled fungal or motifs metal-binding II the Class of I, detection Subsequent domains. conserved ). , .asperellum T. and Trichoderma .virens T. Trichoderma S4 and .I otie rcoin ytae TI)(IS018) 5longiborneol 15 (PIRSF001388), (TRI5) synthases trichodiene 7 contained It ). .arundinaceum T. eeaaye oti n oyo xdsuln yls OC)(fm13249; (Pfam (OSCs) cyclase oxidosqualene of copy one contain analysed here a nyone. only had .guizhouense T. pnn usin bu t vltoayoii.Seiso iieclade Viride of Species origin. evolutionary its about questions opening , S4 n Fig. and hspyoeei itiuinidctsta R5i o a not is TRI5 that indicates distribution phylogenetic This . and 1 .atrobrunneum T. 5 .Tsacsinnmes Scnetprec species each per TS-content numbers, accession TSs ). S1 . .virens T. S4 hra etlnn ytae were synthases pentalenene whereas , .Presilphiperfolan-8 ). and .pleuroticola T. β- lsnhssad22 and synthases ol β- Differently, . .pleuroti T. lsynthases ol S4 ). T. , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. oeo h eoe nlsdi lsl eae pce;wiegene while species; genes related Instead, needed closely are genes. in analyses these further analysed of case, genomes origins any evolutionary the In the of Eurotiomycetes. understand some the better to to belonging order donor in a from (HGT) in transfer gene found the cluster in the proteins (Fig. in synteny homologous encoded for sequences search protein B to the analyses used We BLASTp in 2018). al., gene et Furthermore, the to (Proctor similarity. of associated sequence homolog functionally related no distant showed proteins clade (trichothecene) of Brevicompactum genome TRI the the the of with species proteins trichothecene-producer these of Alignment one and ( transporter of oxygenase efflux upstream one ( located enzymes, oxygenase genes tailoring ), four three of The identification protein. the enabled regulatory systems other in proteins in cluster kb E 21.2 a of in context included genomic the of Assessment is in of centre identity that active (Fig. with acid triad protein the TRI5 NDLFSFYKE showed each and of proteins motif alignments aspartate-rich TRI5 DDSRE/DDSIE of sharing alignment conserved, highly Multi-sequence trichodiene-derivatives. different of as such of presence The of context in genomic pathways the biosynthetic known of Assessment 2. Viride Nevertheless, of domains. species C synthase in from terpene known absent synthase or with were in F synthase clustering which protein polyprenyl TSs chimeric-like, one only I contain as found proteins Class we production named these contains therefore that of group were Most Considering last and clade. The fungi, one. compounds. from than in these TSs more reported produce chimeric have to been species potential not some the some has least least which belonging at diterpenes of species that indole diTS, Some suggesting of Table indole class, 2012). 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SQSs (CPS/KS) “uncharacterized each with synthases as in Copalyl-pyrophosphate/Ent-kaurene cluster present named not is therefore synthase did one were synthase-phytoene they which They but squalene of 1), synthases. conserved 5”, lycopene-phytoene Fig. a reference in contain colour with OSCs (dark-brown neither of PTHR21181:SF13) clade 00494; (Pfam sister domain the from TSs Interestingly, and and C F .gamsii T. eeas on nalteother the all in found also were (Fig. D 2c B ,MjrFclttrSpraiy(F)tasotr( transporter (MFS) Superfamily Facilitator Major ), nw o t blt o ibrli isnhss(ho&Zag21) nadto,w found we addition, In 2015). 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Trichoderma and tri101 Trichoderma .guizhouense T. ugsigti pcei bet rdc sesterterpenes. produce to able is specie this suggesting , R5 otiigbt oan hc shgl iia oophiobolin to similar highly is which domains both containing TR456 tri5 Trichoderma ee yatSAH50(lne l,21)rvae hsgn is gene this revealed 2019) al., et (Blin 5.0 antiSMASH by genes nlsn nte ee htwr ae as named were that genes 6 another enclosing , hc a enarayrpre nother in reported already been has which , .asperellum T. .arundinaceum T. p.ta aentbe ecie strichothecene-producers, as described been not have that spp. C Trichoderma tri5 ,wietetrelctddwsra eeietfidas identified were downstream located three the while ), .citrinoviride T. ugssti eecudb novdi h biosynthesis the in involved be could gene this suggests , 6 ee eel t oeta novmn nun- in involvement potential its reveals genes p.blnigt h iiecae ihconserved with clade, Viride the to belonging spp. tri5 epciey n 7 in 87% and respectively, , u eut eelta hs pce aeat have species these that reveal results our , noeaZ2C rncito atr(F ( (TF) factor transcription Zn2-C6 a encode and Trichoderma .brevicompactum T. , .parareesei T. E E n abncahdae( anhydrase carbonic and ) em ob pcfi to specific be to seems S4 eoe sdhr.Genes here. used genomes .Aayi lorvae set a revealed also Analysis ). , .reesei T. .gamsii T. D hwd7%o amino- of 77% showed .gamsii T. and Trichoderma F A n pce of species and 2a .gamsii T. r rsn in present are , . B tri5 .Pairwise ). F squeries as , C ). species nthe in , D . A A , , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. hc a 3-odu-euae hntefnu a nteros ugsigta inl rmteroots the from signals that in suggesting gene roots, this the of on expression was the fungus triggering the of of when responsible expression up-regulated are 137-fold the was repressed which strongly , roots of the that with extent, contact the (1.58-fold), in occurred of biosynthesis exception sesquiterpene in the biosynthesis with terpene and in genes, in condition, reprogramming expression TS TSs any its affected in of colonization Root most detected control. regulated as differentially conditions which same when the , occurs under biosynthesis PDA on terpene alone on grown regulation differential a roots, whether wheat determine to order In on tested. changes conditions the promoted under that spikes, spikes wheat the in in changes involved that physiological indicate particularly mediate not could are or genes with these Instead, spikes tested. that on conditions indicating the in conditions, interaction of two triple Expression the pathogen. the between of significantly presence/absence the regardless spikes, wheat (Fig. control as alone with (57.9 interacting was fungus the of of genes exception of the transcripts with detect Since 0.33-fold), not to did We (0.02 affected. genes not TS oxidative was of to expression response the in in down-regulated found occur only different might were diterpenes in differences indole occurs of genes biosynthesis in contrast,stress TS that In suggesting of of (2.7-fold), regulation expression respectively). regulated While that 0.6-fold, of stress. indicating (0.3-, oxidative levels down-regulated expression, to transcript response gene in the on manners change changes not opposite did induced medium culture ts9 the MM, to in sucrose grown Nevertheless, 0.9% fungus the of roots. of H Addition wheat cultures (Fig. mM with control liquid 0.5 interaction reference 4-day-old MM in as in sucrose, iii) expression % and TSs 0.9 scenario, the on MM FHB biosynthesis regulating changes a the investigated signals in – firstly environmental in ii) TRI5 We involved the source, and C assess those TS11 of to and presence/absence TS9, order proteins in TS7, in In II and selected TS6, genes Class prenylation. TS5, TS protein encoding of TS4, expression and genes across TS3, precursors excluded distributed TS1, terpene we well – of Thus, group, I functional Class analyses. diverse to our high belonging a TSs represented encoding which genes of 9 genome the on potential in genes genomic TS-encoding high 16 the identified We for driver the in as biosynthesis members terpenoid family TS for of differentiation Functional this, in 3. to pathways According biosynthetic sesquiterpene functions. different unknown two with in trichodiene-derivates novel a of of biosynthesis origin the the suggest findings These n atclry in particularly, and ± Tgam Tgam . S oprdto compared DS) % 4.7 Tgam sal osuppress to able is ts3 Tgam Tgam ts3 ae lc nti cnro oti n,w sesdrltv hne nT xrsinwhen expression TS in changes relative assessed we end, this To scenario. this in place takes Fgra eosre htNsavto eddt eaieyrglt Sgns lhuhsignificant although genes, TS regulate negatively to tended starvation N that observed We . a infiatydw-euae 01fl) hra nu-euainwsosre in observed was up-regulation an whereas (0.1-fold), down-regulated significantly was 05-od.Nvrhls,tems eakbydffrnewsfudi h xrsinof expression the in found was difference remarkably most the Nevertheless, (0.59-fold). a nuae o aso D npeec f1dyodwetselns using seedlings, wheat 1-day-old of presence in PDA on days 3 for incubated was ugsigtepeec fteptoe ol ietyidc hne nisexpression its in changes induce directly could pathogen the of presence the suggesting , i o nuepoietcagsi epn isnhsswe neatdwith interacted when biosynthesis terpene in changes prominent induce not did 3b ts4 .Aayi eeldthat revealed Analysis ). 3a 1.-od.I h aewy diino . MH mM 0.5 of addition way, same the In (18.7-fold). .Teaalblt fCsuc a otatn ffcso Sgn expression. gene TS on effects contrasting had source C of availability The ). ts3 Fgra Fgra ts1 Trichoderma Fgra Tgam and 2 0 nwetsie,w netgtdwehradffrnilepeso fTS of expression differential a whether investigated we spikes, wheat on hc xrsinddntcag (Fig. change not did expression which , 2 nwetsie,weeardcino H yposwsobserved was symptoms FHB of reduction a where spikes, wheat on Tgam Mwt o ocnrto,adM 0 MNC,uigMM using NaCl, mM 200 MM and concentration, N low with MM , ln lns(88.3 plants alone ts11 h ugswsgoni ne ieetsmltdsrs conditions stress simulated different under i) grown was fungus the , Tgam tri5 uigro ooiain Whereas colonization. root during 04fl) iial,adto f20m alt h medium the to NaCl mM 200 of addition Similarly, (0.4-fold). ascae lse in cluster -associated hnclnzn ha ot.I atclr ouainin modulation a particular, In roots. wheat colonizing when ts11 tri5 ts1 Tgam 7 , a lgtyu-euae 14-od when (1.45-fold) up-regulated slightly was Trichoderma , ts5 ts6 Tgam ± (Table ts9 . S,uigsie ncltdwith inoculated spikes using DS), % 1.2 or , ts1 ts9 noigapttv noedT,wsup- was diTS, indole putative a encoding , ts7 nadto,teitrcinwt h roots the with interaction the addition, In . 1 and and .Frgn xrsinsuis efocused we studies, expression gene For ). nayo h odtostested. conditions the of any in . ts11 .gamsii T. ts11 i o change, not did ts7 Trichoderma eeatv when active were ts1 ts11 ts4 3c ts5 hs rncit eenot were transcripts whose , , 2 hc slkl novdin involved likely is which , ts6 .Ti lal niae a indicates clearly This ). O 00-od n nalesser a in and (0.03-fold) xrsin hs results These expression. a lgtyup-regulated slightly was 2 and otecluemedium culture the to ts1 tri5 hc expression which , ts9 p. codn to according spp., ol participate could ts3 Tgam ts6 i o change not did Tgam and and Tgam colonizes ts6 Fgra a on was ts11 Tgam Tgam Tgam ts1 were was tri5 on , . Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. Km hog o,19) eetees h aiblt on ntesrcueo h ciecnr of centre active the pentalenene. of than structure others the sesquiterpenoids synthesize on antibiotics probably fungal found them variability of of terpenoids family the some complex pentalenolactone Nevertheless, that the structurally suggests 1998). of of proteins Yoo, hydrocarbon variety these parent & a the Cheong, generating of (Kim, in (Pinedo biosynthesis fungi contribute the in may sesquiterpenes in polycyclic involved TSs of these range wide Thus, a in of 2008). biosynthesis the al., in role et central a play the longibor- to in this, thought most or to formation that blend According found terpenoid 1996). We in Fitzsimons, the involved actually & terpenoids. of be newly Xue, (D) could of (Cane, residue species biosynthesis mixture Brevicompactum aspartate of product first synthases a the neol or that revealed products Mg have with cyclization which directly TSs motif, interacts D(D/E)HFD on of motif conserved studies metal-binding proteins a crystallography show in and proteins (NDHFD) mutagenesis these conserved of Most partially species. is these in widespread pathways very biosynthetic Mc- is 2016; biosynthesis Mukherjee longiborneol results, & in our (Bansal to pathway According biosynthetic 2009). in culmorin Harris, the & of Alexander, intermediate Viride Cormick, an of 2016), metabolites Mukherjee hypothesized some & We specific that step. synthesizing reported ionization studies sesquiTS an Previous Takahashi, bifunctional by Shinohara, of particular 2009). instead 2005, be step, motifs, al., might clade. DxDTT protonation et TSs and (Nakano a domains HAD-like diTS HAD-like through contain that II cyclization can motif, Class FPP sesquiTS DxDTT some in to a that found leading harbour contain reported work. DxDD proteins also (2016), this HAD-like Koyama the they in These and and of clades. found Osada TRI5-superfamily, variant other TSs the of a of to species is number belonging in which total absent sesquiTS the are in which of rich evolved TSs third particularly HAD-like have the are but almost Viride clades, constitute of which other Species sesquiTSs, the of in an groups present eight constitute are species. results, that clade other our TSs to the Viride According of in of groups absent are some Species which in demands. species. TSs missing environmental specific close are phylogenetically different of they by as species to the shared example, according inventory potential, biosynthetic production terpenoid terpenoid the terpene similar of the their portions despite of Thus, particular within diversity reflect homogeneous and which very nature TSs, is the size TS-family of Although overview complete a inventory. providing most biosynthetic However, genus, fungi. these this in of TSs of ecology of the rich diversity in as the terpenoids considered about and fungi TSs other of the in importance of the found reflects those likely outnumbers as This in clearly 2019). such biosynthesis SMs genome) SMs, terpenoid of per of that complexity genes producers demonstrates and (15-23 diversity analysed species the genomes these the in of in impact found great we a TSs has of number impressive The roots. wheat with DISCUSSION interaction the during fungus the in in enhanced slightly ts9 was biosynthesis triterpene that up-regulated Trichoderma Trichoderma Trichoderma a on ihydw-euae 02-od,tu ugsigarpeso fidl iepn biosynthesis diterpene indole of repression a suggesting thus (0.28-fold), down-regulated highly found was Trichoderma ts6 rtissaigsmlrt ihfna etlnn ytae eeas dnie,wihare which identified, also were synthases pentalenene fungal with similarity sharing Proteins . u sasn nseiso h iiecae ic umrnpouto a o enreported been not has production culmorin Since clade. Viride the of species in absent is but , ogbrelcudb yteie sasll opudo sa nemdaeo unknown of intermediate an as or compound solely a as synthesized be could longiborneol , 15-od and (1.52-fold) S aentytbe hrceie Kbcke l,21) n vial information available and 2019), al., et (Kubicek characterized been yet not have TSs Trichoderma Trichoderma Aspergillus Trichoderma ts11 p.cnptnilypouepresilphiperfolan-8 produce potentially can spp. Trichoderma 25-od.Since (2.58-fold). p.hv ueptnilfrssutreeboytei.W identified We biosynthesis. sesquiterpene for potential huge a have spp. p.(-0gnsprgnm)(eVise l,21;Kbcke al., et Kubicek 2017; al., et Vries (de genome) per genes (2-10 spp. ssac.Tu,w oue ncaatrsn h Sgn arsenal TS-gene the characterising on focused we Thus, scarce. is 2+ .arundinaceum T. Sak,19) n t elcmn a edt anomalous to lead can replacement its and 1997), (Starks, p.hv h oeta osnhsz ogbrel(Bansal longiborneol synthesize to potential the have spp. 8 Tgam Trichoderma ts6 a rdce oecd nSS hssuggests this SQS, an encode to predicted was hnclnzn h ha ot.Interestingly, roots. wheat the colonizing when Trichoderma ewr bet dniyclade-specific identify to able were we , and .brevicompactum T. Trichoderma ned h Sgn inventory TS-gene the Indeed, . β- Trichoderma l hscmon is compound This ol. aeaatdtheir adapted have Site-directed . terpenoid Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. ee eergltdi poiewy nrsos ooiaiesrs.Ascaino xdtv teswith stress oxidative of genes TS Association SMs Similarly, stress. in 2008). oxidative observed Nakajima, to & as response Kawakami, genes, in Jiao, TS ways 2002; of opposite Keller, expression in & the regulated Bok, on were Wilson, functions. effects genes (Calvo, biological contrasting fungi different had other of from source indication showing picture strong C environmental a a of provide different regulated, we Availability differentially under Here, these are in. in grows involved role genes be a fungus TS play could different could the they that that species. processes when genes these about individuating TS of hypothesizing enables numbers organisms, and of gene other frameworks with expression TS interacts high relative when the or for the conditions, driver in the is changes members Assessing family gene of of production differentiation terpenoid functional for potential genomic striking The in in pathways found that metabolic and pathway (TRI biosynthetic Since BGCs sesquiterpenic different interest. two trichodiene-derived agronomic to uncharted potential an with to metabolites novel lead producing could in cluster to present This transferred only species. is likely this extra-cellular cluster were with entire sesquiterpenoid genes the pathway-specific a a these but of suggests production of cluster the the suggests Some within transporter TF functions. a a of of presence presence the The while transporter. regulation, a 2004). of and Altomare, TF role a & Mul`e, enzymes, the Favilla, tailoring (Gallo, about other reported questions Unlike previously some been trichothecenes. have open which leaves of This some found cluster, were TRI proteins entire uncharacterized conserved enzymes other highly that found and We hypothesize including sesquiTS, not we to that clades close thus, shown some phylogenetically 2010); to have are al., restricted studies they et since Biochemical (Lopez-Gallego in 4”, GPP biosynthesis group them. cyclize monoterpene miss-predict to in to able involved leaded monoterpenes are probably of sesquiTS enzymes fungal production these although of work, sequences this in fide analysed in bona genomes reported the confer been among may has found compounds were these monoTS of biosynthesis No diterpene- thus, indole plants, the involved of with Many on being advantages relations Sordariomycetes 2008). ecological symbiotic some al., et in stablish (Saika reported some fungi fungivores been from to producer nematocidal have structures advantages reproductive diterpenes and competitive their indole antimicrobial protecting confer of in their could production and 2017) hand, terpenoids, Hong, other among the & here rare Deng, found are (Tian, diTS Sesterterpenes properties of 2019; diterpenes. number that al., indole low revealed and et the analyses Nevertheless, Chen our 2014; 2019). Interestingly, al., al., et et Zhao (Adelin 2018; genus. species the al., these of et ability in Song the reported indicates 2012; recently al., been terpenoid et have of Miao specific compounds distribution reflects from efficient these likely isolated an of diterpene most first guarantees the This and was Harziandione species, metabolism. Brevicompactum. secondary these and and of biosynthetic primary inventory Harzianum between specific terpenoid in precursors clades precursors the the precursors terpenoid of of of cyclic donors portions as members and act linear in may synthases pathways the FPP and provide GGPP enzymes additional way, of OSC genome most the and in in biosynthesis SQS SQS triterpene single-copy for that required revealed analyses Our ooShv enietfidi ug ShitDnet 04,adtesac viaiiyof availability scarce the and 2014), (Schmidt-Dannert, fungi in identified been have monoTS .virens T. .pleuroti T. Trichoderma Trichoderma Trichoderma tri5 tri5 Cuce ta. 03 nyt,Slsywt,An,&Ysaa,21) No 2019). Yusnawan, & Aini, Sulistyowati, Inayati, 2013; al., et (Crutcher rhlg nnon-trichothecene-producer in orthologs niae htptwyseicSS ol epeeta el ntesame the In well. as present be could SQSs pathway-specific that indicates cnann species, -containing p.frdtreeboytei ssac n o eywdsra within widespread very not and scarce is biosynthesis diterpene for spp. . hs soitosa el(akr&Sot2004). Scott & (Parker well as associations -host .virens T. .gamsii T. Trichoderma Trichoderma .gamsii T. Trichoderma rfl noteTRI5-superfamily. the into fell or Trichoderma ugsigteoii fanovel a of origin the suggesting . tri5 9 tri5 ,w yohsz hsssuT sivle ndifferent in involved is sesquiTS this hypothesize we ), ol eatal nlddwti “uncharacterized within included actually be could p.hv h oeta opouesesterterpenes produce to potential the have spp. nbeneficious in Trichoderma Trichoderma sebde naBCin BGC a in embedded is pce.Hwvr h rsneo nadditional an of presence the However, species. p.(hslet ta. 93,adanumber a and 1993), al., et (Ghisalberti spp. tri5 p.fudi hswr ugssthat suggests work this in found spp. p.b G rmEurotiomycetes, from HGT by spp. Trichoderma Trichoderma em ob ucinlyassociated functionally be to seems tri5 pce isn nthe on missing species .gamsii T. p.nnpoue of producer non spp. Trichoderma ascae G in BGC -associated enclosing , p.In spp. Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. M isnhssi ug a enetnieydmntae,adi a ensgetdta ti induced of is up-regulation it H particular, mM in in that In observed occur suggested 2013). been might Linz, been has diterpenes & has it indole Roze, it of (Hong, and biosynthesis damage demonstrated, ROS that extensively from been fungi prevent has to fungi in biosynthesis SMs in eo San o onigtepoetCU21-4 n h rga Io ototrlFloso the of Fellows Postdoctoral of II R.B. Program and the M-D. Castilla de M.E. and Junta of CLU-2018-04, the grants project; project Salamanca PhD the of this University founding founding for for (Italy) Le´on (Spain) Pisa y of University the acknowledge gratefully We ACKNOWLEDGEMENTS SQS, the of in biosynthesis up-regulation triterpene the and and sesquiterpene sesquiTS on of modulation a expression induces sii the colonization in root observed that suggest effects contrasting the reprogram- addition, significative in a that occurs indicate biosynthesis results ts9 changes Our cause terpene 2018). turn, in of al., which et biochemistry, ming (Contreras-Cornejo plant physiology in changes fungal When substantial the promote 2018). in that Dwivedi, SMs of & in variety (Hidangmayum a TSs releases SMs of including role signals the lecular about by information colonization more Root provide to needed are scenario. this experiments time-course extensive more although spikes, stresses. these overcome to fungus the to advantages In in particular genes confer in TS not shown regulated does negatively been stresses biosynthesis has saline terpenoid it and as starvation production, N 2015). SMs Moulay, in in 2018); changes al., et modulates (Hautbergue stress fungi in metabolism secondary fujikuroi on Fusarium impact considerable a has availability Nitrogen of regulation of types different suggests . This 2011a). al., et tde eemnn hi mato h ln hsooywl rvd oeifrainaotterlsof roles the about information more provide will physiology plant of the interaction the in genes on the these impact in their genes using determining TS by studies addressed of involvement further the be fungal of indicate same must between involvement the relationship that the within beneficial imagine question a can species a promoting we in context, in gene involved this signature sesquiterpenoid In single a 2013). a al., by et governed (Mukherjee pathways genus metabolic in adjustment an in against trichodermin the by from as signals produced such that HA compounds, in suggests phytotoxic gene it of this as of production remarkable, expression was the colonization induce root roots during observed up-regulation strong in expression TS 2013), in 2019, changes al., prominent et of Sarrocco induce genes 2012; not TS al., did of et (Matarese expression pathogen differential with the interacts a by whether production investigated some DON we in reduce Since changes to 2015). induce and al., first spikes et the (Malmierca by produced virulence (HA) its A harzianum and polyketides .brevicompactum T. .arundinaceum T. 68 hog P scnrlnd.Teasneof absence The node. central as FPP through T6085 ugssta otclnzto nue ersini noedtreeboytei ntefnu.In fungus. the in biosynthesis diterpene indole in repression a induces colonization root that suggests , 2 O .cinerea B. 2 otemdu i o induce not did medium the to .graminearum F. ts11 .arundinaceum T. Trichoderma Mlirae l,21) hseapeilsrtshwdffrn clgcldmnsldto led demands ecological different how illustrates example This 2013). al., et (Malmierca taetdteepeso f3oto 0T ee Weane l,21) iial,saline Similarly, 2013). al., et (Wiemann genes TS 10 of out 3 of expression the affected it , , Trichoderma hngoni rsneo %o %scoeo npeec f05m H mM 0.5 of presence in or sucrose 2% or 1% of presence in grown when a on lgtyu-euae.Gn xrsinpten r ihydnmc and dynamic, highly are patterns expression Gene up-regulated. slightly found was tri Aspergillus eeepeso saetdwe rw nda utrswith cultures dual in grown when affected is expression gene nwetsie.Hwvr u eut hwta h rsneo h pathogen the of presence the that show results our However, spikes. wheat on patbnfiilinteraction. beneficial -plant p.(onane l,21) neetnl,adto f09 urs r0.5 or sucrose 0.9% of addition Interestingly, 2016). al., et (Fountain spp. sa niaerltosi novn ihl euae xhneo mo- of exchange regulated tightly a involving relationship intimate an is ak npyooi ciiyadhsacuilrl npatprotection plant in role crucial a has and activity phytotoxic on lacks .gamsii T. tri5 .gamsii T. .gamsii T. xrsinin expression 68 hnclnzswetros onrglto of Down-regulation roots. wheat colonizes when T6085 10 68.Atog ciainof activation Although T6085. tri5 .gamsii T. 68 sal osuppress to able is T6085 tri5 .gamsii T. dlto uat ftefnu.Rslsstrongly Results fungus. the of mutants -deletion .gamsii T. tri5 .gamsii T. xrsini epnet ha pksadthe and spikes wheat to response in expression .brevicompactum T. in 68 ne xdtv tescniin,as conditions, stress oxidative under T6085 .gamsii T. .gamsii T. 68 hnbt ug eeo wheat on were fungi both when T6085 68,dffrnl rmwa observed what from differently T6085, .harzianum T. 68 ihpatros n further and roots, plant with T6085 Trichoderma .gamsii T. 68 cuswe h fungus the when occurs T6085 68 and T6085 .gamsii T. .graminearum F. .cinerea B. tri5 68 n ha plants, wheat and T6085 tri5 Tjrn ta. 2011a), al., et (Tijerino Bae,Mhmd & Mohamed, (Bualem, ooie h ot,it roots, the colonizes nteboytei of biosynthesis the in sal ed othe to leads usually .brevicompactum T. .cinerea B. 68,suggesting T6085, ee ikdto linked genes 2 O ts9 2 nwheat on (Tijerino suggests .gam- T. while , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. yagnm iigapoc:assetreesnhs from (2013) synthase H. sesterterpene Oikawa, a & approach: 594–597. K., mining genome Gomi, a A., by Minami, (2019). R., W. Zhang, Chiba, . . . 368–374. H., : Guo, fungus 86 J., endophytic , Xu, the S., from Li, sesquiterpenoids Y., new two Chen, H., Li, S., Chen, genus 4867–4877. the : of 77 fungi , filamentous (2011). in Proctor, Guti´errez, S. S.P., genes McCormick, biosynthesis N.J., . . Alexander, . M.R., Hermosa, R.H., M.G., Malmierca, R.E., Cardoza, mutagenesis site-directed (1996). by B.C. region Fitzsimons, aspartate-rich & conserved Q., Xue, D.E., Cane, (2015). B. Moulay, & B., harzianum Mohamed, B., (2008). Bualem, B. Tudzynski, (2019). in & cluster T. P., gene Weber, biosynthetic Hedden, Microbiology . . gibberellin F., . the Gong, of S.Y., M.C., rization Lee, Rojas, N., C., pipeline. Bomke, Ziemert, mining genome R., metabolite Villebro, secondary W81–W87. the K., : to Steinke, updates S., 5.0: antiSMASH Shaw, K., Blin, (2016). G. Vannacci, & wheat S., Sarrocco, of G., sequence Piaggeschi, genome A., Zapparata, R., Baroncelli, Vannacci, . . . P`e, M.E., A., Zapparata, Announcements E., Bertolini, L., (2015). Fiorini, G. G., Piaggeschi, R., J. Baroncelli, (2016). Ouazzani, K. . P. . . Communications Mukherjee, Product P. & Retailleau, R., B.I., Bansal, Iorga, 55–61. G., : Arcile, 97 , M.T., Martin, C., (2014). Servy, E., Adelin, Expression and (2001). Structure H. Gene from Noguchi, cyclase & oxidosqualene Y, of Takagi, mutations K., (2005). Naito, I., D. Abe, Posada, & R., evolution Zardoya, F., Abascal, REFERENCES av,AM,Wlo,RA,Bk .. elr ..(2002). N.P. Keller, & J.W., development fungal Bok, and R.A., metabolism Wilson, A.M., Calvo, aristolochene (2002). to Society, R.K. farnesylpyrophosphate Chemical of Allemann, conversion & Synthase-mediated P.R., lochene Ashton, M.J., Calvert, eoeAnnouncements Genome . iylcadttaylcdtree rma from diterpenes tetracyclic and Bicyclic Bioinformatics . against rf hl-eoeSqec ftebooto agent biocontrol the of Sequence Whole-Genome Draft 4:5325–5339. : 74 , e00647-15. : 3 , 2 11636–11641. : 124 ortscin´ereaBotrytis rcoem asiT6085 gamsii Trichoderma 1:1934578X1601100. : 11 , 1:2104–2105. : 21 , 52:67–73. : 1522 , e01747-15. : 4 , . irbooyadMlclrBooyReviews, Biology Molecular and Microbiology . dnicto flc n ucinlcaatrzto ftrichothecene of characterization functional and loci of Identification sa ora fPatPathology Plant of Journal Asian ehlsoimceues iciiae ipyiaAt BA - (BBA) Acta Biophysica et Biochimica caerulens. Cephalosporium h epni isnhsstoktof toolkit biosynthesis terpenoid The rmsn icnrlaeto uaimHa lgton Blight Head Fusarium of agent biocontrol promising a , Trichoderma rcoem.ApidadEvrnetlMicrobiology Environmental and Applied Trichoderma. 11 rcoem koningiopsis Trichoderma paeoamnhtcl.ApidadEnvironmental and Applied manihoticola. Sphaceloma rcoin ytae rbn h oeo h highly the of role the Probing synthase. Trichodiene rtet eeto fbs-tmdl fprotein of models best-fit of selection ProtTest: aiiyiflec pnatvt of activity upon influence Salinity Biochemistry . oeua lnn,epeso,adsite-directed and expression, cloning, Molecular seglu clavatus Aspergillus ybotof symbiont emceeAi rdc fteAristo- the of product a is A Germacrene dnicto fohooi synthase F ophiobolin of Identification . rcoem azau 67.Genome T6776. harzianum Trichoderma 158–166. : 9 , eainhpbtensecondary between Relationship 5:12369–12376. : 35 , au act.Phytochemistry baccata. Taxus uli cd Research Acids Nucleic A729. ora fteAmerican the of Journal . he e iepnsand diterpenes new Three 66 . slto n characte- and Isolation rai Letters Organic rcoem.Natural Trichoderma. iogncChemistry Bioorganic 447-459. : rf Whole- Draft Trichoderma 5: 15 , 47 , Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. omtblmc,mvn oada nertdsrtg o h icvr ffna eodr metabolites secondary fungal (2018). of I.P. discovery the Oswald, for & strategy O., integrated Reports Puel, Product an Natural (2004). toward L., moving Debrauwer, M. metabolomics, E.L., Lorito, to Jamin, & T., I., Hautbergue, Chet, A., symbionts. Viterbo, plant avirulent C.R., opportunistic, Howell, G.E., Harman, fungus filamentous Microbiology, (2018). the A.J.M. (2010). of Driessen, of O. ring & performance Gascuel R.A.L., the Bovenberg, & Assessing Guzm´an-Ch´avez, R.D., W., phylogenies: Zwahlen, Hordijk, Maximum-Likelihood F., estimate M., to Anisimova, 3.0 PhyML methods V., and Lefort, algorithms J.F., New Dufayard, S., Guindon, Raton eds. RJ, neux (1993). E.L. Ghisalberti, (2012). R.J. catalysis. Peters, complex & of view R.B., Honzatko, Y., Gao, 11–20 in gene (2004). homologous C. Altomare, synthase & M., Mul`e, Favilla, (2016). A., G., expression B. Gallo, gene Guo, biosynthetic . metabolite . . secondary V., and Kumar, enzyme, M.K., Pandey, L., Yang, of S.N., Responses Nayak, P., Van- Bajaj, . J.C., . Fountain, . A., Scartazza, growth. Micro-Tom P., cv. Picciarelli, tomato M., (2016). Curadi, G. L., nacci, Mariotti, (2018). L., L.,Guglielminetti, C. Fiorini, Altomare, strain. biocontrol & multi-target Ku- A.F., a of Logrieco, . . . V.C., atrobrunneum Trichoderma Y., Liuzzi, Miao, F., D., Fanelli, Yang, L., Atanasova, fungus J., mycoparasitic . . (2018). Zhang, . C.P. K., C.A., bicek, Chenthamara, Uchima, genus I.S., S., fungal Druzhinina, Amillis, important medically G., and Aguilar-Osorio, industrially the A., in Wiebenga, (2017). R., I.V. Riley, Grigoriev, R.P., Vries, De fungus C.M. biocontrol Kenerley, the & S., in Zeilinger, P.K., Mukherjee, R., (2013). Schuhmacher, A., Parich, F.K., Crutcher, armyworm Biochemistry fall and the Biology of (2018). rate J. sitism Larsen, & Mac´ıas-Rodr´ıguez, Gonz´alez-Esquivel,C.E., Alarc´on, E., A., L., Del-Val, H.A., Contreras-Cornejo, (2016). J. Ecology Microbiology Larsen, FEMS & of E., functions Mac´ıas-Rodr´ıguez, del-Val, H.A., L., Contreras-Cornejo, . , 15–18. uaietreecyclase, terpene putative A ytmtcBiology Systematic . Trichoderma seglu flavus Aspergillus 9 iatv aua rdcs eeto,ioainadsrcueelucidation. structure and isolation detection, products: natural Bioactive 2768. : rcoem harzianum Trichoderma Trichoderma asv aea rnfro ee noigpatcl aldgaigezmst the to enzymes wall-degrading cell plant encoding genes of transfer lateral Massive rcoem ies uglGntc n Biology and Genetics Fungal virens. Trichoderma 5:147–173. : 35 , aua rdc Reports Product Natural p.adterscnaymtbltsi h hzshr:itrcin ihplants. with interactions rhizosphere: the in metabolites secondary their and spp. 2 196–202. : 122 , oprtv eoisrvashg ilgcldvriyadseicadaptations specific and diversity biological high reveals genomics Comparative ( .harzianum T. 2:fiw036. : 92 , eeto n slto fbocientrlpout.I:Clgt M Moly- SM, Colegate In: products. natural bioactive of isolation and Detection rcoem azau.PyilgcladMlclrPatPathology Plant Molecular and Physiological harzianum. Trichoderma rcoem atroviride Trichoderma eiilu chrysogenum Penicillium ln Soil Plant ooiaiesrs r eae ofna eeomn euao,antioxidant regulator, development fungal to related are stress oxidative to 9:307–321. : 59 , pdpeafrugiperda Spodoptera M Genomics BMC rmispatascae hosts plant-associated its from vir4 srsosbefrteboytei fvltl epn compounds terpene volatile of biosynthesis the for responsible is , aueRvesMicrobiology Reviews Nature 0 351–366. : 400 , pce ope)IE 0:isgtit h eei endowment genetic the into insight 908: ITEM complex) species 67 ouae ope eaoi ewr ostimulate to network metabolic complex a modulates T6776 9:1153. : 29 , 9:662. : 19 , 12 az otascae ugs nrae h para- the increases fungus, associated root maize a , epni ytaesrcue:as a incomplete far so a structures: synthase Terpenoid yisntrlenemy natural its by scl atr o aua products natural for factory cell as seglu.Gnm Biology Genome Aspergillus. slto n hrceiaino trichodiene a of characterisation and Isolation rnir nMicrobiology in Frontiers . LSGenetics PLOS . 4:377–393. : 84 6:67–77. : 56 , eoi hrceiainof characterization Genomic apltssnrni.Soil sonorensis. Campoletis 4:e1007322. : 14 , Trichoderma 8:28. : 18 , R rs,Boca Press, CRC , rmgenomics From 7 rnir in Frontiers . : 2048. Ecological Enginee- species 5: 65 , . Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. n Genomics in and cluster gene biosynthesis carotenoid (2017). T.F. Wang, Linnemannst & S.Y., Tung, Y.C., Chuan,g clusters. C.L., Chen, reesei C.H., derma Huang, (2000). W.C., Y. Li, Ebizuka, 6816–6824. & : into 122 K., synthase , lupeol Masuda, engineering M., synthases: Shibuya, T., Kushiro, . . . Genomics J., Zhang, BMC B., (2019). Henrissat, G., I.S. Manganiello, Druzhinina, K., Chenthamara, A.S., Steindorff, C.P., Kubicek, of (2011). style life I.G. Thon, ancestral I.S., Grigoriev, the Druzhinina, D.A., . . Martinez, . V., M., Seidl-Seiboth, A., Herrera-Estrella, C.P., approach. Kubicek, formation bond (1998). carbon-carbon J. consecutive Yoo, via dl-pentalenene & of synthesis J.H., Cheong, S., Kim, usability. and (2013). F. performance D.M. in Bidard, Standley, provements & . . . K., A., Katoh, (2014). Margeot, S. R., Hunter, Biofuels Koszul, for Y., . nology Vicq, . . D., C., Poggi-Parodi, McAnulla, (2017). L., Baudry, W., E., Li, Jourdier, M., classification. Fraser, function protein H.Y., genome-scale Chang, 5: D., InterProScan Binns, P., Jones, 212–219. : 285 (2008). , T. Nakajima, and & production A., Kawakami, F., Jiao, Col- Chemistry & Guti´errez, Biomolecular S., J.R., & Hanson, Organic arundinaceum. L., Lindo, (2018). R.E., I.G. Cardoza, lado, J., Moraga, I., Izquierdo-Bueno, . (2019). E. Yusnawan, from & compounds L.Q., nic Aini, L., Sulistyowati, A., Inayati, (2013). J. metabolism. Linz, secondary & of L., Roze, S.Y., Hong, (1992). A.E. in Desjardins, synthase trichodiene & T.M., Hon, review. a (2018). stresses: P. abiotic Dwivedi, & A., Hidangmayum, eds. MG, Tuohy I, (2014). Druzhinina of E. RS, metabolites Monte, antimicrobial & Guti´errez, S., and M.B., Rubio, lism R.E., Cardoza, R., Hermosa, rxmt iainsaodn n oprsno two of comparison and scaffolding ligation Proximity itcnlg o Biofuels for Biotechnology opeegnm eune eetidcdpitmtto,adpriinn fCZm gene CAZyme of partitioning and mutation, point repeat-induced sequence, genome complete n,P,Pao . enadzMr´n . uznk,B,&Aao,J (2002). J. Avalos, & Fern´andez-Mart´ın, B., M., Tudzynski, Prado, R., P., ¨ ons, 6 593–602. : 267 , Tri 0:485. : 20 , 0:151. : 10 , eeepeso by expression gene eeac ftedlto fthe of deletion the of Relevance rcoem ies nentoa ofrneo ilg n ple cec (ICOBAS) science applied and biology on conference International virens. Trichoderma ibrlapulicaris Gibberella Trichoderma ora fPamcgoyadPhytochemistry and Pharmacognosy of Journal Toxins vlto n oprtv eoiso h otcommon most the of genomics comparative and Evolution 683–702. : 5 , itcnlg n ilg fTrichoderma of Biology and Biotechnology 0:170. : 10 , oprtv eoesqec nlssudrcrsmcprsts as mycoparasitism underscores analysis sequence genome Comparative Trichoderma. uaimfujikuroi Fusarium . uaimgraminearum Fusarium eoeBiology Genome oeua ilg n Evolution, and Biology Molecular . AF utpeSqec lgmn otaeVrin7 Im- 7: Version Software Alignment Sequence Multiple MAFFT oeua ln-irb Interactions, Plant-Microbe Molecular β- xdtv tesrltdtasrpinfcosi h regulation the in factors transcription stress-related Oxidative mrnsynthase amyrin slto n eedsuto fthe of disruption gene and Isolation aia ylzto fNAiiiyiie .Hgl efficient Highly 4. N-Aziridinylimines of cyclization Radical ln epne to responses Plant Tatri4 n ia P cml ,HreaEtel ,Upadhyay A, Herrera-Estrella M, Schmoll GP, Vijai In: 6:2955–2965. : 16 , 13 ffcso ieetcro ore ntrichothecene on sources carbon different of Effects 2:R40. : 12 , h genes the : eei h eodr eaooeof metabolome secondary the in gene Bioinformatics rcoem reesei Trichoderma nlqi culture liquid in ora fteAeia hmclSociety Chemical American the of Journal . Trichoderma carB niuglatvt fvltl orga- volatile of activity Antifungal uainlsuiso triterpene on studies Mutational -SN 2278–4136. E-ISSN: 0:772–780. : 30 and 0:1236–1240. : 30 , leir 125–137. Elsevier, , ESMcoilg Letters Microbiology FEMS . aR.MlclrGenetics Molecular carRA. p.adtertleac to tollerance their and spp. tan genomes strains 5 Synlett 249–256. : Tox5 Trichoderma eodr metabo- Secondary 98:981–982. : 1998 , eeencoding gene Trichoderma Biotech- . species Tricho- A . Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. oer,S,Xog . isn .. vnv,Y,Sih .. a,GS,&Matsuda, & G.S., May, M.L., Smith, Y., Ivanova, W.K., . Wilson, Q., (2009). Xiong, S.P.T. S., Lodeiro, (2001). 2 the T.D. and Schmittgen, PCR quantitative & K.J., Livak, (2016). Lipids J. synthase of Chappell, squalene Biology & of S.A., domain Bell, functional X., specific Zhuang, T.D., Niehaus, K.B., Linscott, amec,MG,MCrik .. adz,RE,Aeadr .. ot,E,&Guti´errez, & E., Monte, N.J., Alexander, (2014). R.E., Cardoza, S. S.P., McCormick, Hermo- M.G., Malmierca, I.G., Biology Collado, and (2013). S.P., Genetics Guti´errez, S. Fungal McCormick, arundinaceum. . Trichoderma N.J., . . Alexander, R., R.E., sa, Cardoza, M.G., Malmierca, 1103–1118. : 17 , (2015). in Alexander, Guti´errez, role R.E., S. tential Cardoza, I., . . Izquierdo-Bueno, . S.P., McCormick, N.J., (1996). J., C.P. Barua, M.G., Kubicek, Malmierca, & by antagonism B., mycoparasitic Bodo, of 6382–6385. S., target Rebuffat, major V., C. a Farkas, Schmidt-Dannert, M., & Lorito, isomers M.D., geometric pyrophosphate Distefano, farnesyl D., of cyclization Abate-Pella, S.A., (2010). Agger, F., Lopez-Gallego, (1987). L. Willmitzer, & tissues J., plant Schell, J., Logemann, ishsi . ikl . kd,M,Ae . ikca,JS (2017). J.S. Dickschat, & from synthases I., sesterterpene Abe, chimeric M., two Okada, of J., rization Rinkel, (2012). T., N.Y. Mitsuhashi, Ji, & G., from Wang, X.L., diterpenes Yin, harziane X.R., Liang, F.P., Miao, production. culmorin for (2016). required 136–141. (2009). I. synthase L.J. Abe, Harris, longiborneol & a . . N.J., . Alexander, M., S.P., Okada, McCormick, elucidation T., and method, Mori, sponge biosynthesis. M., crystalline its and Hoshino, of spectroscopy NMR S., by determination Lee, structure T., Astellifadiene: Mitsuhashi, Y., (2012). Matsuda, G. Vannacci, & V., Trichoderma between Seidl-Seiboth, 98–106. interactions S., blight: Gruber, head S., Fusarium Sarrocco, F., Matarese, mycotoxigenic and (2010). mould F. green Matarese, by produced peptaibols hosts their of with dynamics and V´agv B., profile Leitgeb, Diversity A., Szekeres, C., Urb´an, Tyagi, T., P., Marik, fungi antagonized and plants by strain rai Letters Organic eqiepn ytae o4adCp from Cop6 and Cop4 synthases Sesquiterpene nltclBiochemistry Analytical . rdcino rcoin by trichodiene of Production ortscinerea Botrytis grcsbisporus Agaricus rtsainlboytei n eaoimi h ahgncfungus pathogenic the in metabolism and biosynthesis Protostadienol 1:1241–1244. : 11 , 81:1049–1057. : 1861 , Fusarium neadeChemie, Angewandte icnrlo uaimHa lgh oeua neatosbetween interactions molecular Bligth: Head Fusarium of Biocontrol rcoem pce.OgncLetters Organic species. Trichoderma -CT h hss nvriyo ia ia Italy. Pisa, Pisa, of University Thesis, PhD . naoitcatvt n ln eec priming. defence plant and activity antagonistic method and N eeac ftihteee nfna hsooy irpinof Disruption physiology: fungal in trichothecenes of Relevance vlapnld rdcinby production aspinolide ovel niomna Microbiology Environmental . luou srau.Ceity&Biodiversity & Chemistry ostreatus. Pleurotus 6 16–20. : 163 , Methods . 128 rcoem harzianum Trichoderma 5879–5882. : iciiae ipyiaAt BA oeua n Cell and Molecular - (BBA) Acta Biophysica et Biochimica . nlsso eaiegn xrsindt sn real-time using data expression gene relative of Analysis rcoem azau.Junlo Bacteriology of Journal harzianum. Trichoderma 5:402–408. : 25 , 14 eiilu.Ceity-AErpa Journal European A - Chemistry Penicillium. ChemBioChem . n mycotoxigenic and 3:22–33. : 53 , ornscinereus Coprinus mrvdmto o h slto fRAfrom RNA of isolation the for method Improved ple n niomna Microbiology Environmental and Applied 4:3815–3817. : 14 , CLM1 lestepreto fti biocontrol this of perception the alters rcoem arundinaceum Trichoderma 7:2628–2646. : 17 , ly,M,...Keis .(2017). L. Kredics, . . . M., ¨ olgyi, Trichoderma of 1:1093–1106. : 11 , bouecngrtoso unique of configurations Absolute uaimgraminearum Fusarium uaim Microbiology Fusarium. aayi rmsut and promiscuity Catalytic : niomna Microbiology Environmental pce ninteractions in species 4:e1700033. : 14 , ehnsi characte- Mechanistic elwl ytei is synthesis wall Cell seglu fumigatus Aspergillus apn kingdom- a Mapping icnrlof Biocontrol Trichoderma ihapo- a with encodes 7 : 178 , tri5 5 : 158 , 6: 76 , 3: 23 , in Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. i,B,Mtua . oi . kd,M,Qa,Z,Mtuah,T,Wkmt,T,&Abe, & T., Wakimoto, swapping. T., domain Mitsuhashi, by Z., synthase Quan, sesterterpene a M., into Okada, T., Mori, (2015). Gu- I. Y., . Matsuda, . . B., L Qin, Lindo, A.M., fungi Stanley, entomopathogenic R.E., and pathogenic Cardoza, H.S., Kim, . (2018). S.P., . ti´errez, . S. McCormick, P., R.H., Proctor, (2010). Pecheur, A.P. Le alignments. Arkin, large & M., for P.S., trees Choquer, Dehal, M.N., B., Price, Dalmais, J.M., Pradier, (2008). topathogen in pathway C.M., M. mevalonate of Wang, Viaud, branches sterol C., and dolichyl Pinedo, the of enzymes Gene of reesei. activity Trichoderma (2014). on effects J.S. P., contrasting Kruszewska, Zembek, has B., . Antosiewicz, . . S., G´orka-Nie´c, Perli´nska-Lenart, Graczyk, U., eds. S., W., Pi A, Varma lsyk, DK, Choudhary Plants In: in Resistance management. disease (2016). Systemic in H.M. role Paikrao, possible and & istry S.R., 405–426. Patil, York, A.S., New Patil, Dekker, Marcel Mycology. (2004) Industrial B. of Handbook Scott, & eds. E.J., BN, Parker, Johri SK, (2019). Das P. T, I. Satyanarayana Mukherjee, Abe, In: Sustainability diversity. & geno- . P.D., and . chemo- . Sherkhane, K., S., Nakagawa, Pachauri, T., Mori, S., Hoshino, sis. T., Mitsuhashi, aphidicolin of Y., (2016). biosynthesis Matsuda, T. in M., Sassa, skeleton Okada, . diterpene . . unusual Y., Society an Kamiya, Chemical of American H., formation Kawaide, for S., responsible Ohashi, H., Toshima, (2001). T., Toyomasu, H., Oikawa, D., feed Bhatnagar, and T.E., food Cleveland, of (2015). L., contamination G.A. B. Losada, Payne, Scott, N.D., . Fedorova-Abrams, . . & J., Yu, G.A., W.C., Payne, Nierman, B.L., Pritchard, B.J., of Monahan, A., (2009). Koulman, M.J., (2005). Nicholson, T. Hoshino, 1016–1018. & : T., 8 , Dairi, T., Sato, H37Rv3377c T., Okamura, C., Nakano, (2009). the T. from Hoshino, cyclase, & (2013). C.M. C., Kenerley, Nakano, & M., Schmoll, A., Herrera-Estrella, Trichoderma B.A., Horwitz, P.K., Mukherjee, 10053–10057. eiilu paxilli Penicillium ora fteAeia hmclSociety Chemical American the of Journal dnicto ftoaarmboytei eelc in loci gene biosynthesis aflatrem two of Identification lnn n ucinlepeso fcN noigaphidicolan-16 encoding cDNA of expression functional and Cloning eoebsddsoeyo nupeeetdcciainmd nfna etrepni biosynthe- sesterterpenoid fungal in mode cyclization unprecedented an of discovery Genome-based ortscnra C hmclBiology Chemical ACS cinerea. Botrytis nuuulciei iepn ytaefrom synthase diterpene chimeric unusual An noe h iepn yls o rdcn h aiaeskeleton halimane the producing for cyclase diterpene the encodes eerhi h eoeEra Genome the in Research igpr,Srne,441-456. Springer, Singapore, , yoatru tuberculosis Mycobacterium vlto fsrcua iest ftihteee,afml ftxn rdcdb plant by produced toxins of family a trichothecenes, of diversity structural of Evolution oeuiaeterfunction their elucidate to eqiepn ytaefo h orda isnhtcgn lse ftephy- the of cluster gene biosynthetic botrydial the from synthase Sesquiterpene 4 114–122. : 544 , LSONE PLoS eoesqec of sequence Genome 2 5154–5155. : 123 , igpr pigr 69–102. Springer, Singapore , eoeAnnouncements Genome . vrxrsinof Overexpression . noedtreeboytei nacmctu fungi ascomycetous in biosynthesis Indole-diterpene e9490. : 5 , LSPathogens PLOS . hrceiaino the of Characterization nulRve fPhytopathology of Review Annual . 3 genome. H37 3 10011–10018. : 138 , ple n niomna Microbiology Environmental and Applied . seglu flavus Aspergillus neadeChemie Angewandte 791–801. : 3 , 15 erg20 Trichoderma atre2–ApoiaeyMaximum-Likelihood Approximately – 2 FastTree mrclavariecolor Emericella e00168-15. : 3 , ChemBioChem, 4:e1006946 : 14 , eeecdn anslprpopaesynthase pyrophosphate farnesyl encoding gene seglu flavus Aspergillus Rv3377c RL35,asri htcue aflatoxin causes that strain a 3357, NRRL eodr eaoimin metabolism Secondary eodr eaoie:Terbiochem- Their metabolites: secondary 2 1690–1693. : 128 , eepout yeBditerpene type-B a product, gene . 10 irba iest nEcosystem in Diversity Microbial β- 1:105–129. : 51 , 2060–2071. : n t ucinlconversion functional its and lsnhs:akyenzyme key a synthase: ol Ceia Communications .Chemical yoatru tuberculosis Mycobacterium irba-eitdInduced Microbial-Mediated n eaoi engineering metabolic and ora fthe of Journal . 5:7469–7481. : 75 , . n nZ(ed), Z An In: Trichoderma : Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. hrs,M,Hra,GE,Msor,F (2010). agents. F. biocontrol Mastouri, fungal G.E., Harman, M., Shoresh, biosynthesis. astellolide (2016). in Y. involved Koyama, machinery & H., biosynthetic Osada, S., Takahashi, Y., synthases Shinohara, plant . the . . with B.F., common Dunican, in D.J., determinants Spakowicz, (2015). K., S.A. Jefferson-George, T., Strobel, Sasaki, T., Berbasova, J.J., Shaw, 19–61 Springer, eds. J, Yanofsky,mann & (2014). C. V.E.A., Schmidt-Dannert, Russo, W., Zhou, Chemistry M., Biological Schumacher, (1994). F.R., C. Lauter, (2013a). T.J., G. Vannacci, Schmidhauser, & conditions. A., field Ritieni, under A., Blight Agriculture Head Moretti, Fusarium G., of Pachetti, control L., logical Moncini, Duf- Blight. S., . Head . Sarrocco, Fusarium . of S., biocontrol Domenichini, the T., in Plainchamp, traits R., perspectives. 2020. Bernardi, future I., M. and Vicente, resne, P., art Esteban, of S., State Sarrocco, cereals: staple in reduction (2019). 701 risk P. mycotoxin Battilani, for A., approach Mauro, G. S., Vannacci, & Sarrocco, A., Moretti, A., Ritieni, G., Pathology Pachetti, L., Moncini, (2013b). F., Matarese, S., eds. Sarrocco, Garc´ıa-Estrada C, E, Rhizomicroorganisms Sansinenea (2019). Promoting MS, Growth V. Reddy Plant C, Sharma, of Keswani (2008). HB, & Singh B. N., In: overview. Scott, Rialch, & R., E.J., Salwan, fungi Parker, filamentous C., in Young, diversity chemical M.J., indole-diterpene Nicholson, S., Saikia, Sciences of Academy National the (2001). D.W. Christianson, sporotrichioides & Fusarium D.E., Cane, M.J., Rynkiewicz, (2003). J.P. Huelsenbeck, models. & F., Ronquist, agent (2008). biocontrol I.G. the Collado, & of R., species Hernandez-Galan, from R.F., Guerrero, J.L., Reino, (2019). K. eds. Surapathrudu, I, & A.C., Solanki, derma M.K., Solanki, S., Rai, (2014). C. Schmidt-Dannert, Report & Products C.M., Flynn, M.B., Quin, . ln elhUdrBoi Stress Biotic Under Health Plant p. urn nesadnsadftr uloso oeua ehius nAsr A Mahmood RA, Ansari In techniques. molecular on outlooks future and understandings Current spp.: Bioinformatics, icnrlo uaimha lgtb pk plcto of application spike by blight head Fusarium of Biocontrol 1:19–27 : S1 . 6:95–100. : 86 , hrceiainof Characterization itcnlg fIornis dacsi iceia Engineering/Biotechnology, Biochemical in Advances Isoprenoids. of Biotechnology . 1:1449–1473 : 31 , ta optto n ha otedpyimof endophytism root wheat and competition Straw 6 12060–12066. : 269 , 19 nulRve fPhytopathology of Review Annual dnicto fafna ,-iel ytaefrom synthase 1,8-cineole fungal a of Identification rvdsmcaitcifrne ntetreecciaincascade. cyclization terpene the on inferences mechanistic provides 1572–1574. : . al-2 isnhsso epni aua rdcsi ug.I:Shae ,Bohl- J, Schrader In: fungi. in products natural terpenoid of Biosynthesis 8:13543–13548. : 98 , rcoem.Pyohmsr Reviews Phytochemistry Trichoderma. h hteesnhs eeof gene synthase phytoene the , pigr igpr,129–160. Singapore, Springer, , Phytopathology rae :Bysa hlgntcifrneudrmixed under inference phylogenetic Bayesian 3: MrBayes ora fBooia Chemistry Biological of Journal . igpr,Srne,87–111. Springer, Singapore, , 16 s fcmeiiefiaetu ug sa alternative an as fungi filamentous competitive of Use iatv oaiemtbltsof metabolites volatile Bioactive yooia Research Mycological . nue ytmcrssac n ln epne to responses plant and resistance systemic Induced 8:21–43. : 48 , cetfi Reports Scientific icnrlo ln ahgn nSustainable in Pathogens Plant of Biocontrol ( submitted rvrigtefna terpenome fungal the Traversing tutr ftihdeesnhs from synthase trichodiene of Structure rcoem asi ora fPlant of Journal gamsii. Trichoderma rcoem asiT6085 gamsii Trichoderma dnicto fanvlsesquiterpene novel a of Identification ersoacas.TeJunlof Journal The crassa. Neurospora 89–123. : 7 , icnrlptnilof potential Biocontrol ) Hypoxylon 32865. : 6 , 1 184–199. : 112 , 9 8511–8526. : 290 , eodr metabolites Secondary eodr Metabolites Secondary h eei ai for basis genetic The p ihspecificity with sp. Trichoderma rceig of Proceedings Toxins, Singapore Natural . suseful as Tricho- An : 11 Bio- : Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. ag . ornn,K,Kphnky . gce,RK,Aaaoa . hnhmr,K., Chenthamara, producer L., cellulase (2015). Atanasova, the I.S. R.K., of Aghcheh, Druzhinina, A., (2012). . Kopchinskiy, . M.G. . K., Feng, Pomraning, . D., . . Yang, X.Q., Xie, in 483. entomopathogenicity S., : fungal Zhang, 2 of Z.L., evolution , Wang, the P., on perspectives Zheng, J.J., nomic S.H., Ying, Espino, G., E.M., Xiao, Niehaus, L., Studt, Pathogens (2013). K.W., B. Bargen, gen Tudzynski, von C.M.K., . . . Sieber, P., Wiemann, approach. Maximum-Likelihood a using families (2001). protein N. Goldman, & S., Whelan, (1998) G.E. enzymes. Schulz, & (2007). K.U., C. Kenerley, Wendt, . . & . I., H., Chiet, Y., Li, virens Brotman, Trichoderma A., M.J., Wiest, A., Barbetti, Viterbo, R., Pathology Marra, Plant Molecular E.L., and Physiological Ghisalberti, (2008). M. K., Lorito, Sivasithamparam, gene. F., synthase diphosphate Vinale, (1998). copalyl the Y. of Kamiya, characterization & and T. cloning H., Sassa, Kawaide, B., . . Tudzynski, . T., Sciences Dairi, of Academy W., National Mitsuhashi, the R., Niida, A., Kaneko, (2007). Gutier- M., Tsukahara, . seedlings . T., tomato . Toyomasu, on J., and Aleu, genes M.G., biosynthetic Malmierca, trichodermin J., the Moraga, (2011b). R.E., S. Cardoza, rez, R., Hermosa, Hermosa, A., . . Tijerino, . J., Aleu, F., in Vicente, activity antimicrobial M.G., Malmierca, J., (2011a). Moraga, R. R.E., Cardoza, A., Grant, Tijerino, (2017). K. . . Hong, . & Drugs L., X., Marine Ryder, Deng, V., G., Perera, Tian, R., Winsbury, genomics. K., from Cocq, insights Le (2013). B., M. Harris, D.J., Studholme, (2018). thase. N.Y. (1997). Ji, C.M. & Starks, X.L., Yin, fungus F.P., algicolous marine Miao, the from S.T., Fang, Y.P., Song, (2007). D.W. diphosphate. Christianson, from & Synthase D.E., Aristolochene Cane, of L., Costanzo, Di E.Y., Sishova, Fusarium Science uiocn r isnhszdb nuuulcieadtreesnhs nfungi in synthase diterpene chimera unusual an by biosynthesized are Fusicoccins Structure e1003475. : 9 , netgtn h eeca risof traits beneficial the Investigating Biochemistry, vrxrsino h rcoin ytaegene synthase trichodiene the of Overexpression uiuo eelcmlxrglto fscnaymtbls n oe metabolites novel and metabolism secondary of regulation complex reveal fujikuroi 5:229. : 15 7 1815–1820. : 277 , vrxrsino the of Overexpression :127-133. 6 , lctpatdfneresponses defence plant elicit oe oefor role novel A rnir nPatScience Plant in Frontiers tutrlbssfrcci epn isnhssb oac -p-rsoohn syn- 5-Epi-aristolochene tobacco by biosynthesis terpene cyclic for basis Structural rcoem rvcmatm uglGntc n Biology and Genetics Fungal brevicompactum. Trichoderma rcoem esi eoeAnnouncements Genome reesei. Trichoderma 6:1941–1951. : 46 eihrn h rpi eoe eoewd nlsso h iepatho- rice the of analyses genome-wide genome: cryptic the Deciphering seglu terreus Aspergillus eoesqec n noainof annotation and sequence Genome rcoem harzianum Trichoderma 0 3084–3088. : 104 , Trichoderma rcoem rvcmatmtri5 brevicompactum Trichoderma eea miia oe fpoeneouindrvdfo multiple from derived evolution protein of model empirical general A . speodboytei:mnfl hmsr aaye ysimilar by catalyzed chemistry manifold biosynthesis: Isoprenoid 2:80–86. : 72 , h ilgclatvte fssetreodtp ophiobolins. sesterterpenoid-type of activities biological The oeua ln Pathology, Plant Molecular . rcoem hamatum Trichoderma n vlto ftmltsfrtecciaino farnesyl of cyclization the for templates of evolution and 258. : 4 , eodr eaoie nteitrcin ihplants. with interactions the in metabolites secondary 17 oeua ilg n Evolution and Biology Molecular X-5. ibrli isnhssin biosynthesis Gibberellin Toxins . ora fNtrlProducts Natural of Journal tri5 nrae rcoemnpouto and production trichodermin increases rcoem parareesei Trichoderma euei asaa cetfi Reports Scientific bassiana. Beauveria 1220–1232. : 3 , urn Genetics Current e00885-15. : 3 , D2frssanbearclue- agriculture sustainable for GD12 ee ffc nteepeso of expression the on Effect gene: iepnsadsesquiterpenes and Diterpenes 8 h 8e etiosfrom peptaibols 18mer The 737–746. : X 8:285–296. : 48 , rycytlstructure crystal -ray ibrlafujikuroi: Gibberella 4:234–240. : 34 , 8:691–699. : 18 , 1:2553–2559. : 81 , rceig of Proceedings . h ancestor the , PLOS . Ge- Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. l uhr elr ocnito interest. of conflict no its declare to authors contributed All authors the and all Interests E.M. and of G.V., manuscript, and Conflict the data, the Bernardi wrote analysed R. I.V. M.E.M-D., M.E.M-D interpretation. Baroncelli, and revision. R. R.B. data I.V., S.S., to I.V., research. contributed the experiments. R.H. designed the Baroncelli. performed G.P. R. and and R.H. S.S., E.M., G.V., CONTRIBUTION AUTHORS Trigam1 code of sequence in ( available XM are as: XM S1 NCBI in Fig. available are Information analyses Supporting and ( synthases in Terpene Institute (NCBI). Genome shown Joint the numbers of databases accession public protein synthases Terpene (2014). ACCESIBILITY DATA J.W. Xu, & of X.R., culture Jing, submerged gene. a Y.L., synthase in He, acids M.F., ganoderic individual Ren, of S.L., Ji, J.S., Zhou, 1588–1597. : 14 , (2015). Y. asperellum choderma Zhang, & (2019) L., C. fungus Zhao, Wang, the & of C., strain Shao, coral-derived C., Reports soft Wang, a T., from Shi, diterpenes L., Yang, (2016) D., P.K. Zhao, Mukherjee, & R., Bansal, – derma S., Sar- Gruber, . . S., . Zeilinger, G., Vannacci, L., Malfatti, Mu˜noz, pathogen. I., Vicente plant S., (2017). Somma, S. D., rocco, Lio, Da A., Zapparata, (2005). Phytopathology, N. of Keller, & J.H., Yu, (2001). B. Scott, from & cluster E., gene Telfer, indole-diterpene Scott, L., McMillan, . C., . . Young, G.T., Biology Bryan, and R.D., Genetics Johnson, Fungal G., lolii. Spangenberg, K., (2006). Shields, B. S., Felitti, C.A., Young, ts11 2500. ( 024550105.1 ,XM ), 13345. : 9 , hmsr et genomics. meets Chemistry .gamsii T. ope eecutrfridl-iepn isnhssi h rs endophyte grass the in biosynthesis indole-diterpene for cluster gene complex A 1807 ( 018800875 9898. iceia niern Journal Engineering Biochemical eoeAnnouncements Genome 3:437–458. : 43 ts4 eoesequence Genome 1o rmtn uubrgot ne atstress salt under growth cucumber promoting on Q1 ,XM ), 68 NB G rjc ceso ubrJD0000)uigthe using JPDN00000000) number accession Project WGS (NCBI T6085 tri5 1801 ( 018800313 .Ncetd euneof sequence Nucleotide ). β- uui eeacsinnmesof numbers accession gene tubulin euaino eodr eaoimi lmnosfungi. filamentous in metabolism secondary of Regulation 3:679–693 : 43 , eiilu ail.MlclrMicrobiology Molecular paxilli. Penicillium ffcso hsht ouiiainadpyoomn rdcinof production phytohormone and solubilization phosphate of Effects uglBooyReviews Biology Fungal fFsru graminearum Fusarium of 2596 ( 024549662 ts5 e01209-17. : 5 , ,XM ), JGI . 0:178–183. : 90 , β-τυβυλιν 1809 ( 018800491 n h ainlCne o itcnlg Information Biotechnology for Center National the and ) 18 aoem lucidum Ganoderma rcoem harzianum Trichoderma ts9 ,XM ), 0:74–90. : 30 , eewsetatdfo h hl genome whole the from extracted was gene oeua lnn n eei nlsso an of analysis genetic and cloning Molecular ts6 TM14(TC501,amycotoxigenic a 56091), (ATCC 124 ITEM .gamsii T. ,XM ), 1859 ( 018805995 ora fItgaieAgriculture Integrative of Journal . . 2595 ( 024549058 eodr eaoimi Tricho- in metabolism Secondary yteoeepeso fsqualene of overexpression the by 68 sdfrgn expression gene for used T6085 . 9:754–764. : 39 , ts1 oetpyooi harziane phytotoxic Potent ,XM ), nacdaccumulation Enhanced XS-20090075 ts7 ,XM ), 1818 ( 018801080 nulReview Annual Neotyphodium JGI Scientific . 018806700 protein ts3 Tri- ), Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. c) b) a) T.brevicompactum T.brevicompactum T.arundinaceum T.arundinaceum T.guizhouense T.guizhouense T.asperellum T.asperellum B. bassianaB. bassianaB. T.asperellum T.asperellum T.gamsii T.gamsii T.gamsii T.gamsii ARSEF2860 ARSEF2860 Phylogeny CBS433.97 CBS433.97 NJAU4742 NJAU4742 IBT40841 IBT40837 IBT40841 IBT40837 TR356 TR356 T6085 T6085 A5MH A5MH 0.53 0.93 0.53 0.58 0.58 0.7 0.86 0.7 0.9 0.87 * 0.88 0.93 1 0.8 0.88 0.86 0.92 0.78 0.7 0.93 0.99 0.98 0.98 1 0.9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 C 21.2 kb 21.2 Presilphiperfolan Predicted function Indole Longiborneol Oxidosqualene E Pentalenene Trichodiene - Diterpene diterpene Squalene Uncharacterized Uncharacterized group group 2 Uncharacterized Uncharacterized Uncharacterized group group 1 Uncharacterized Uncharacterized GGPP - 8 FPP β Chimeric 19 - ol GGTases GGTases synthases synthases synthases synthases synthases synthases synthases HAD synthases synthases cyclases FTases group group group group - - like like 1 2 4 3 5 Enzyme type Enzyme Chimeric Class I Class I Class Class Class I Class Bifunctional Bifunctional DXDDand DEXXE Class Class II Bifunctional Bifunctional and DDXXEDXDTT Class I Class I Class I Class Class Class I Class I Class I Class I I I I I I I – – – – – – – – – – – – – – – – – TS TS DDXXX TS TS [D/E]XX[D/E] TC DDSRE / DDSIE TC TC DDQFD / DDPAA TS TS DDIRE TC TC DD(M/V/L)FD TC TC DDHFD / DEHFD / NDHFD PT and DDYLD DDVQD PT (IPS) DDIMD and DDXXD TS TS DXXXE PT D[D/E]XX[D/E] PT DEWGE PT DEXXV/ DAXXV PT DDMTI/ DDMTH TS TS DDXXD PT (IPS) DDVID and DDMLD PT/TC PT TC / DCISE DCTSE - TC D[D/E]XXDand/or D[D/E]XXD ** Aspartate - rich rich motif Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. b) a)

-DDCt Relative expressionTítulo del eje (2 ) -DDCt 2.00 1.00 4.00 0,50 1,00 2,00 4,00 0.50 Relative expression (2 ) Tgam 16,00 32,00 32.00 16.00 0,03 0,06 0,13 0,25 0,50 1,00 2,00 4,00 8,00 0.06 0.13 0.25 8.00 0.03 2.00 0.50 1.00 4.00 tri5 TRI5 tri5 ts1 Sucrose Sucrose 0.9% *** ts3 Series2 TS1 ts1 ts4 *** Tgam Tgam + Fgra TS3 ** ts3 ts6 * *** H 2 Series1 O ts7 *** 2 0.5 mM TS4 ts4 ts9 ts11 TS5 ts5 * Series3 N starving TS6 * ts6 ** c)

-DDCt

Relative expression (2 ) TS7 ts7 128,00 256,00 16,00 32,00 64,00 Tgam Series4 NaCl 200 mM 0,02 0,03 0,06 0,13 0,25 0,50 1,00 2,00 4,00 8,00 ...... *** 20 *** t TS9 s9 tri5 *** ** ts11 TS11 ts1 * *** ts3 * ts4 *** Tgam Tgam + wheat 0.9% 0.9% sucrose 0.5 0.5 mM H 200 NaCl mM N starvation ts5 * ts6 ** 2 O ts7 2 ts9 *** ts11 *** Posted on Authorea 3 Aug 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159646770.00457953 — This a preprint and has not been peer reviewed. Data may be preliminary. Trigam1|6072 Trigam1|8345 Trigam1|9898 Trigam1|2917 Trigam1|4065 Trigam1|3927 Trigam1|5139 Trigam1|3208 Trigam1|340 Trigam1|5367 Trigam1|4947 Trigam1|1824 Trigam1|9843 Trigam1|162 Trigam1|4742 Trigam1|5596 JGI Id. Name TRI5 TS10 TS11 TC4 TC2 TC1 TC5 TC3 TS3 TS4 TS7 TS5 TS9 TS8 TS6 TS1 21 bifunctional HAD bifunctional pentalenene synthase pentalenene oxidosqualene trichodiene synthase trichodiene Putative functionPutative uncharacterized 5 uncharacterized 4 uncharacterized squalene synthase squalene GGPPsynthase FPP transferase FPPsynthase indole indole GGTase GGTase sesquiTS sesquiTS sesquiTS diTS II cyclase I - like like