South America: an Invaluable Source of Yeast Diversity Lacking a Specialized Culture Collection Diego Libkind Lab. Microbiología Aplicada y Biotecnología, Instituto de Investigaciones en Biodiversidad y Medioambiente (INIBIOMA), Universidad Nacional del Comahue – CONICET Bariloche, Argentina. Presentation overview Survey of Yeast-preserving Culture Collections in South America Contributions from Patagonia (Argentina) to yeast… Biodiversity Taxonomy Biogeography Ecology - photobiology Biotechnology Basidiomycetous yeasts Survey: Yeast-preserving Culture Collections in South America World Federation for Culture Collections (WFCC) database Latinoamerican Federation for Culture Collections (FELACC) 2009 Survey Organized by the Argentinean Association for Microbiology (AAM) AAM 2009 Survey COUNTRY N° CC With yeasts COUNTRY N° CC With yeasts Brazil 60 6 Argentina 18 6 Argentina 11 3 Brazil 5 1 Venezuela 3 ni Venezuela 1 0 Colombia 2 1 Uruguay 2 1 Chile 1 ni Ecuador 1 1 TOTAL 77 10 TOTAL 27 9 ni: no information available Results from both databases differ considerably Survey: Yeast-preserving Culture Collections in South America Country Collection Origin N° Yeast strains % Yeast Brazil UFMG, Belo Horizonte Environ., industrial, other 15000 70 Argentina CRUB, INIBIOMA-UNCo Environ., Type strains 3000 100 Ecuador CLQCA, CNIB Environ., Other 1800 100 Brazil URM, Recife WFCC Various 1600 18 Argentina IMR-M Clinic, environ. 1426 57 Argentina EEA, INTA Wine, other 1330 100 Argentina IDEPA-UNCo Wine, cider industries, other 1098 100 Argentina Dmic, Malbran Clinic, environ. 425 23 Argentina CCCE, CEREMIC Clinic, other 402 28 Brazil INCQS, Fiocruz WFCC Various 277 28 Argentina CCMFCA Wine industries 211 33 Ecuador Hospital Vozantes Clinic 120 9 Brazil CBMAI Altern. Research Collect. Environ., various 100 5 Brazil UFPEDA; Pernambuco WFCC Various 75 13 Argentina LPSC, Spegazzini Inst. International collections 50 5 Brazil IGESALQ, U. Sao Paulo WFCC Various 50 6 Brazil IPT, S.P. WFCC Various 34 8 Brazil CBMAI WFCC Various 30 2.5 Argentina CCM-A, UBA WFCC n.a. 21 2 Colombia CMPUJ, Univ. Javeriana Clinic 20 1 Argentina BNM, BsAs WFCC various 2 0.5 Sources: 2009 AAM survey/ WFCC database/ Collection´s oficial web site / curator´s personal communication Survey: Yeast-preserving Culture Collections in South America Country Collection N° Yeast strains Year Described Not yet described TOTAL sp. nov. sp. nov. sp. nov. Brasil UFMG, Belo Horizonte 15000 1994 35 50 85 Argentina CRUB, INIBIOMA, Bche. 3000 2002 12 20 32 CLQCA, CNIB, Quito Ecuador http://www.cnib.com.ec/CLQCA 1800 2006 1 15 16 TOTAL 19800 (30-40% ID) 48 85 133 South America Universidade Federal DPS, Ontario, Canada Dr. André Lachance de Minhas Gerais, ICB NCYC, Norwich, England Dr. Carlos Rosa Dr. Steven James CREM, Lisboa, Portugal Dr. Jose Paulo Sampaio Universidad Católica de Quito Dr. Javier Carvajal CRUB UNCo CONICET Instituto de Investigaciones MABB en Biodiversidad y Medio ambiente Microbiología Aplicada y Biotecnología Dr. Maria de van Broock Bariloche – Patagonia Argentina Contributions from Patagonia to yeast… taxonomy (Basidiomycota) Taxonomy (basidiomycetesSubphylum) : Agaricomycotina (ex. Hymenomycetes Class) Ecology Biotechnology Subphylum: Ustilaginomycotina (ex. Ustilaginomycetes Class) Subphylum: Pucciniomycotina (ex. Urediniomycetes Class) Bauer et al., 2006. Mycol. Progress Contributions from Patagonia to yeast… taxonomy (Agaricomycotina) Agaricomycotina (D1/D2) Holtermanniales New order: Holtermanniales Cryptococcus ? New genus: Holtermanniella 4 new combinations for Cryptococcus species New species: Holtermannia takashimae Holtermanniales H. takashimae HB982 T - FM246501 H. takashimae SEG-5-6 - FN430734 A (ITS) H. takashimae VTT C -04546 - FM246503 H. takashimae SEG-9-3 - FN430732 62 H. takashimae CBS 10506 - FJ473372 56 H. takashimae VTT C -04547 - FM246504 B clone BF-OTU102 - AM901778 82 H. takashimae CRUB1154 - FM246502 H. takashimae HB 981 - FM246505 C 68 H. takashimae DVBPG 4685 - FM882240 99 H. festucosa VKM Y -2930 T - AY462120 0.02 76 H. mycelialis CBS 7712 T - AF408417 99 62 H. nyarrowiiCBS 8805 T - AF400697 H. wattica CBS 9496 T - FJ473373 Holtermannia corniformis CBS 6979 T- AF410472 FilobasidiumglobisporumCBS 7642 T - AF444336 Wuczkowski et al., IJSEM, In press Contributions from Patagonia to yeast… taxonomy (Agaricomycotina) Agaricomycotina (D1/D2) Holtermanniales Description of new genus/species Cryptococcus ? Review status of Mrakia curviuscula New combination for Crypt. huempii Polyphyletic Cryptococcus restricted to 3 orders: Tremellales, Trichosporonales and Filobasidiales Cystofilobasidiales (D1/D2) 100 Mrakia robertii 100 Mrakia gelida Mrakia/Mrakiella Mrakiella niccombsii (Margesin & Fell, 2008) 74 82 Mrakiella aquatica Tausonia pamirica 42 100 Guehomyces pullulans 50 Itersonilia perplexans 100 | Udeniomyces pannonicus | Udeniomyces pseudopyricola 100 | PR74 sp. nov. (4 isolates) PR7 100 | Cystofilobasidiales yeast 35 | Mrakia curviuscula 100 Cryptococcus huempii Xanthophyllomyces dendrorhous 0.01 Contributions from Patagonia to yeast… taxonomy (Agaricomycotina) New concept: Ecoclade (Gadanho & Sampaio, 2009) Exclusive combination of phylogenetic and ecological Extreme environments: Acidotolerant yeasts and highly characteristics. resistant to toxic metals pH Toxic metals (1.8-2.7) Volcanic origin (Russo et al., 2010) New species: Holtermannia takashimae Copahue volcano (Argentina) Anthropogenic origin Abandoned mines (Portugal) Filobasidiales ARD: Acid Rock Drainage (D1/D2) Contributions from Patagonia to yeast… taxonomy (Ecoclade) New concept: Ecoclade (Gadanho & Sampaio, 2009) Other example of an Ecoclade: Psychrophylic yeasts in the Microbotryomycetes Origin: Himalayas and Alps Margesin et al., 2007 Shvaji et al., 2008 Contributions from Patagonia to yeast… biogeography Xanthophyllomyces dendrorhous (Cystofilobasidiales) North hemisphere South hemisphere Astaxanthin Slime fluxes Betula, Cornus, Fagus trees Cyttaria spp., exclusive parasite of Nothofagus trees Contributions from Patagonia to yeast… biogeography Relationship between X. dendrorhous lineages and hosts CRUB 1149 AF000000 lake Ilon Nothofagus dombeyi U96853 ) CRUB 853 AF000000 Nothofagaceae Nothofagus pumilio U96851 B A CRUB 1490 AF000000 Cyttaria Argentina CRUB 1491 AF000000 Nothofagus ( Nothofagus antarctica U96849 KBP 2604 AF000000 Betula papyrifera AF432067 + Carpinus betulus + Betula verrucosa KBP 2607 AF000000 (ITS) Betula pendula AY352332 + Betula populifolia Host CBS 7918T AF139628 Russia Betulaceae trees CBS 6938 AF139632 Finland + Italy (Weber & Davoli, 2005) Alnus japonica AY352314 Betula ATCC 24228 AF139631 Alaska + Germany (Weber et al., 2006) (ITS) Dendrorhous Xanthophyllomyces + EEUU (Fell et al., 2007) ATCC 24230 AF139630 Fagus crenata AY232918 ATCC 24261 AF000000 Japan Fagus japonica AY232914 CBS 5908 AF000000 Alnus CBS 9090 AF444488 Fagaceae Cornus eydeana AY243787 ? PYCC 4172 AF000000 Fagus (Fell et al., 2007) Cornus officinalis AY530921 CBS 5905 AF139629 Fagus Japan Type of P.rhodozyma Corneaceae Cornus controversa AY530918 ATCC 24229 AF000000 Ulmus americana AF174640 ATCC 24261 AF000000 Japan Cornus Fitzroya cupresoides AY178418 Cyst. macerans CBS 2206T 10 10 Contributions from Patagonia to yeast… biogeography Phylogeny based on crtI, crtS and idi genes of X. dendrorhous CRUB1614 Cyttaria hariotii on Nothofagus dombeyi CRUB1613 Cyttaria hariotii on Nothofagus dombeyi 91 CRUB1620 Cyttaria hariotii on Nothofagus dombeyi N. dombeyi CRUB1492 Cyttaria hariotii on Nothofagus dombeyi CRUB1490 Cyttaria hariotii on Nothofagus dombeyi 100 63 CRUB0853 Cyttaria hariotii on Nothofagus dombeyi Nothofagaceae 57 CRUB1621 Cyttaria hariotii on Nothofagus antartica CRUB1151 Cyttaria hariotii on Nothofagus pumilio N. pumilio/antarctica 82 CRUB1149 Cyttaria hariotii on Nothofagus pumilio 85 CRUB1617 Cyttaria hariotii on Nothofagus antartica KBP2604 Betula sp. 51 KBP2607 Betula sp. Betulaceae CBS7918 Betula verrucosa 54 CBS6938 Betula sp. ATCC24228 Betula papyritosa 63 63 ATCC24230 Betula tauschii 72 CBS5908 Alnus japonica ATCC24261 Betula maximowicziana 89 CBS5905T Fagus crenata Fagaceae CBS9090 (same as CBS5905T) PYCC4172 (same as CBS5905T) ATCC24229 Cornus brachypoda 100 ATCC24201 Cornus brachypoda Corneaceae 10 Contributions from Patagonia to yeast… taxonomy (Basidiomycota) Taxonomy (basidiomycetesSubphylum) : Agaricomycotina (ex. Hymenomycetes Class) Ecology Biotechnology Subphylum: Ustilaginomycotina (ex. Ustilaginomycetes Class) Subphylum: Pucciniomycotina (ex. Urediniomycetes Class) Bauer et al., 2006. Mycol. Progress Contributions from Patagonia to yeast… taxonomy (Pucciniomycotina) Polyphyletic genera: Rhodotorula Microbotryomycetes Sporobolomyces Ballistoconidia Bensingtonia forming yeasts Cystobasidiomycetes Pucciniomycetes Atractiellomycetes Agaricostilbomycetes Carotenogenic (red) yeasts Bauer et al., 2006 UV Contributions from Patagonia to yeast… Taxonomy (Mycosporines) MIC Mycosporines (MYC) in yeasts: Chemotaxonomical value mAU Rhodotorula minuta UV spectrum 1 1200 310 nm UV + PAR 0,75 1000 Single compound with 800 maximum absorbance AU PAR 0,5 HPLC at 310 nm 600 0,25 Control 400 0 250 300 350 400 nm 200 0 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 Retention time Mycosporine-glutaminol-glucoside (MGG) UV Contributions from Patagonia to yeast… Taxonomy (Mycosporines) MIC Mycosporines (MYC) in yeasts: Chemotaxonomical value Species-specific characteristic (Red yeasts) Rhodotorula? Erythrobasidiales and Cystobasidiales MYC, Mycosporine production; PAC, Production of