UMR 1136 INRA/UHP Tree-microbes Interactions “Physiology and functional genomic of transporters” team Fungal pathogen : Melampsora larici-populina Populus trichocarpa

Melampsora larici-populina Haustorium formation Adapted from Mengden et al., Biotrophic interfaces Infected leaves At a cytological level

Molecular and metabolic dialogue. Entrainment of plant metabolism by phytopathogenic fungus 1.Sensing nitrogen or carbon availability 2.de-repression of specific metabolic pathways (transporters; enzymes) 3.Plant defence responses are taken up through AA permeases. 4.Plant invertase produce depolymerized sugars that both partners can use. - Which role(s) transporters play in molecular dialogue, in exchange of nutrients ? - Which transporteurs are key molecular markers for the interaction? Divon and Fluhr, 2007

Toward an overview of Melamspora “transportome”…… © INRA - TCDB: Transport Classification DataBase families -TransportDB 1.Retrieve proteins from DB 1.Analysis - Blastp - search for IPR, PFAM domains - search by KOG classification And………manual curation (tblastn for some families)

Plant pathogensSaprophytes Plant Symbionts Animal pathogens

Increased genetic potential for transmembrane channel proteins (water; solute as small carbohydrates (e.g. glycerol), urea, nitrate, CO 2, H 2 O 2, ions)

Family specific to plant-pathogen fungus (at least basidiomycetes). ascomycetes???

What makes a pathogen an obligate highly specialized pathogen?

1. Less ability to transport and likely utilize mineral nitrogen coumpounds.

2. an increased genetic potential for peptide uptake...

EuGene |Mellp1: EuGene Mellp1:96051 estExt fgenesh2 pg.C |Mellp1:11... PGTG |predicted protein EuGene |Mellp1: EuGene Mellp1:88574 EuGene |Mellp1: estExt fgenesh2 pg.C 20111|Mellp1: EuGene |Mellp1: PGTG |conserved hypothetical p... YPR194C (Opt2p) Ca (OPT6) Ca (OPT5) Ca (OPT4) Ca (OPT3) Ca | (OPT2) e gw Mellp1:34758 PGTG |sexual differentiation p... YJL212C (Opt1p) Ca (Opt1p) Scpo (Isp4p) EuGene |Mellp1: EuGene | Mellp1:84873 PGTG |conserved hypothetical p... Ca (OPT7) EuGene |Mellp PGTG |predicted protein EuGene | Mellp1:88746 PGTG |hypothetical protein EuGene |Mellp1: EuGene |Mellp1: PGTG |predicted protein PGTG |predicted protein EuGene |Mellp1: fgenesh2 pg |Mellp1: PGTG |predicted protein estExt fgenesh2 pg.C Mellp1:11... PGTG |predicted protein PGTG |predicted protein EuGene |Mellp1: PGTG |conserved hypothetical p... PGTG |predicted protein PGTG |predicted protein PGTG |predicted protein PGTG |predicted protein PGTG |predicted protein PGTG |predicted protein estExt fgenesh1 pg.C Mellp1:77... PGTG |conserved hypothetical p... ScYgl114wp Ca (OPT8) EuGene Mellp1:85440 PGTG |conserved hypothetical p... Tetrapeptide signature Mlp genes in tandem Mlp genes Phylogenetic analysis of the OPT family NJ method, Poisson correction, 1000 bootstrap replicates (MEGA4)

L. bicolor :2 P. chrysosporium:6 C. cinerea: 2 C. neoformans: 0 U. maydis: 2 P.graminis: 14 M.larici populina: 8

Phylogenetic analysis of the APC family YAT ACT LAT AAAP NJ method, Poisson correction, 1000 bootstrap replicates (MEGA4)

Lacbi1: CC1G Af Agp2 YBR132C AGP2 CNAG CNAG NCU YOR348C PUT4 En PutX NCU Phchr1: Lacbi1: Lacbi1: Lacbi1: Lacbi1: Hc GAP1 UM CNAG CNAG estExt Genewise1Plus.C Mellp1:... Ufa AAT1p e gw Mellp1:34045 Ufa AAT2 e gw Mellp1:37139 estExt Genewise1Plus.C Mellp1:... fgenesh1 pg.C scaffold Mell... UM NCU PENCH Mtr UM Lacbi1: CC1G Am AAT1 Phchr1: CNAG UM e gw Mellp1:36162 e gw Mellp1:36184 Homology P... estExt fgenesh1 kg.C Mellp1:72... Ufa AAT3p EuGene Mellp1:90273 Homology... e gw Mellp1:38039 e gw Mellp1:41039 Lacbi1: CC1G Phchr1: YPL265W DIP5 UM CNAG CNAG NCU YNL268W LYP1 YNL270C APL1 YEL063C CAN1 Lacbi1: CC1G Phchr1:4184 Af LYP Lacbi1: Phchr1: YFL055W AGP3 UM Af Gap1 PENCH GAP NCU NCU UM NCU03509 Naap1 CC1G Lacbi1: Lacbi1: Gm YLL061W MMP1 YPL274W SAM3 YGR191W HIP1 YKR039W GAP1 YOL020W TAT2 YBR069C TAT1 YCL025C AGP1 YDR508C GNP1 YBR068C BAP2 YDR046C BAP3 NCU UM Phchr1:4672 UM fgenesh2 pg Mellp1: fgenesh2 pg Mellp1: CC1G Phchr1: Lacbi1: CNAG Lacbi1: Lacbi1: SCHPO CNAG UM NCU CNAG CNAG En GabA UM YNR056C BIO5 YGL077C HNM1 Lacbi1: CC1G Phchr1: Phchr1:27552 CNAG UM estExt Genewise1Plus.C Mellp1:4... NCU00765 AAP2 Lacbi1: Lacbi1: CC1G Phchr1: NCU e gw Mellp1:40209 e gw Mellp1:37702 Lacbi1: Phchr1: YDL210W UGA4 UM Phchr1: Phchr1:5862 Phchr1:26909 Lacbi1: Lacbi1: estExt fgenesh2 pg.C Mellp1:11... YHL036W MUP3 Lacbi1: CC1G Phchr1:37473 UM CNAGJEC CNAG estExt fgenesh2 pg.C Mellp1:11... Af MUP NCU CC1G CNAG Lacbi1: Lacbi1: Lacbi1: Phchr1: NCU NCU YGR055W MUP1 Lacbi1: CC1G CC1G Phchr1:38005 Lacbi1: CC1G EuGene Mellp1:84909 Phchr1: NCU UM CNAG UM Phchr:185 UM NCU AAP1 CNAG CNAG Lacbi1: CC1G Phchr:45006 CNAG estExt Genewise1.C Mellp1:41988 NCU YKL146W AVT3 UM YNL101W AVT4 Phchr1:6774 CC1G UM gw Mellp1:30092 CNAG Lacbi1: CC1G UM gw Mellp1:24482 CNAG NCU YEL064C AVT2 YBL089W AVT5 YER119C AVT6 YIL088C AVT7 estExt fgenesh2 pg.C Mellp1:11... UM CNAG Phchr: Lacbi1: CC1G Uromyces fabae genes (PIG, permeases) Mlp genes tandem

- OPT : example of how evolution has shaped Mlp genome to suit specialized pathogenic lifestyle. - Role of nutrition in the host-pathogen relationship with respect to host restriction and evolution towards obligate pathogenesis Conclusion Small number of transporter-encoding gene models EST support