ARTICLE 39 et al s. str 2

et al$%+% s. str have led to the

et al. $%%& $%+% Key words: distribution ecology R ;' Lactarius * #' et al. $%++ 1 ; IMA · VOLUME 5 · NO 1: 39–49 NO 1: 5 · · VOLUME FUNGUS IMA s. lat sequences Lactarius Lactarius '< et al. $%+% < Multifurca and 0 Molecular investigations in =&%> related Stubbe ' " #9 * $%+%;4(' al$%%)567$%%8* et ' 4D $G et al$%%0 ) in Togo ) in Togo Russulales , et al $%+% Lactarius subbaliophaeus #" (F + Plinthogali ( s. str. ; et al.$%%&' Piperites Lactarius 0%' et al $%%) * 'DE and L. griseogalus

+$ Lactarius s. str. +G/' s. str. et al$%%1

;9 Lactarius 0

Lactarius EK4*#ECD55G1*9@%%%E* 4*!O*?C59C"9KCCRF)8&%)G&CE .'KL*L+$GL* (*E*4U*6;4**9+&)%C* 4 *< . FK 5 E* (U*! K(!;V ; C" ; ; ? "! " .FO \ " (West Africa): phylogeny and a new species described a new species phylogeny and Africa): (West better circumscribe the genus 4('!;FC =59 the delimitation of the genus from the delimitation of the genus Abstract: *L+1+15 ;=9?N Article info:F?8.$%+0='?$&'$%+0=L?&C$%+0 notably L. baliophaeus © 2014 International Mycological Association © 2014 International Mycological conditions: distribute and transmit the work, under the following are free to share - to copy, You Attribution: 2 G 1 + 45 C The genus The genus 0 Non-commercial:No derivative works:Any of the above conditions can be waived if you get For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Nothing in this license impairs or restricts the author’s moral rights. permission from the copyright holder. . / Savolainen use of modern methods (molecular tools) such as those "4('4(' " ( The diversity of biological organisms in a site can be assessed The diversity of biological organisms in a ! vulnerable ecosystems are urgently needed to monitor these " #" INTRODUCTION " ( ?+%11@&A$%+0%1%+%1 VOLUME 5 · NO. 1 Maba et al.

< et al. $%++ subgenera PiperitesRussulariaPlinthogali "Lactarius ' tenellus'4DG@81*#L. kabansus'<@@9 $%>Lactarius s.str. +8@EV(;L. griseogalus (R. Nicholson ' 0% 179 DC ;

ARTICLE #Xet al$%%G<6/$%+% +GEF L. tenellus L. kabansus U D6/+@8& ; Russula Amanita Tomentella Cantharellus Xerocomus Boletellus Boletus Microscopy Pulveroboletus and Tylopilus .< LactariusR 6/$%+%Cet al.$%+GFVC VUC " ' C et al $%+G L <6*$%%+4D6E$%%8# Lactarius s. lat. et al.$%%84 et al.$%+%*et al$%+$Cet al. material collected in similar ecosystems in the neighbouring $%+G *#Xet al$%%G' ! / ' Lactarius and < 6 / $%+% 9 9 'Lactarius s.lat. 4Det al. $%%$4et al.$%%$C$%+%<6/ DNA extraction, sequencing, and PCR $%+%*et al$%+$(VUC and Lactarius have been #4('4(' '# et '4DG@81 '<@@9+8@ al.$%%84 et al.$%+%*et al.$%+$ Cet al.$%+G;!;F .1))%%2;" 4('!;F+!;F$1&F !;F+. 2 the Basidiomycota !;F0* E 6 * /''+@&@; +@@G/+@!;FLactarius s. str. +% 9VUC9 8 Russula + Termitomyces 9 +=.+;+' ' et al. $%%$ ; .R V('V(' C(L E 'X . Sequence analyses and molecular # VUC9 phylogenetic inference F $%%8 .*('et al +@@8 ; R " Lactarius s. str. ! *( ! *Lactarius s. str. ( F 4 !(F4= * et al. ; L. afroscrobiculatus L. atro-olivinus $%%& V(' ?AAAA L. miniatescens and L. tenellus4D K(!;VDXet al.$%%1'et al.$%+% 6 E $%%8 C $%+% < 6 / $%+% ' ;!;F!;F+ " !;F$ 9 Lactarius ; s. str; *V8$1$0F$%+G= " 7$%%1O!;FG@9 9 ; "9 L. subbaliophaeus /9Lactarius the Togo Lactarius $0 +$ Russula G 9Termitomyces $"Agaricus $Hymenagaricus+=.+ ;.C MATERIAL AND METHODS 59!(F9C'..; 8+G%D6;$%%&=$8F$%+G Specimen sampling and loan of material ' ;; C et al. $%+G .

40 IMA FUNGUS Lactarius s. str. in Togo ARTICLE

Fig. 1. * C" 5 Lactarius ; '*&%>E*K(!;V(U*! V('\

VOLUME 5 · NO. 1 41 Maba et al.

Table 15!;F4(' Species Collection numbers Country ENA accession numbers Lactarius afroscrobiculatus '4D01@9 Togo 7E@+8G88 Lactarius kabansus C4+G$ Togo 7E@+8G8) Lactarius kabansus '<@@9+8@ ] 7E@+8G@% ARTICLE Lactarius saponaceus C4G@% Togo 7E@+8G8& Lactarius subbaliophaeus C4+%% Togo 7E@+8G8$ Lactarius tenellus C4+0@ Togo 7E@+8G8G Lactarius tenellus '4DG@8 D 7E@+8G&@ Lactarius C4G@+ Togo 7E@+8G&% Lactarius miniatescens C4+1+ Togo 7E@+8G80 Lactarius miniatescens C4+88 *. 7E@+8G81 U$+)& Togo 7E@+8G&0 U$G)$ Togo 7E@+8G&1 '4D0G+1 Togo 7E@+8G&G U$001 Togo 7E@+8G@+ U+&$$ Togo 7E@+8G&$ Lactilfuus foetens C4+1% Togo 7E@+8G&+ C4G&@ Togo 7E@+8G&) Termitomyces C4G&& Togo 7E@+8G&& Russula congoana C4+$@ Togo 7E@+8G&8

O00!;F4(' ; L. purpureus L ( E belonging to the subgenus Piperites E*8%%;C" '" 5 C5 CVE' 1$ F C4+%% Lactarius (Tamura et al $%++ ^ $%+G LactariusC E ; # A E;#_E_!E4E #LF C4+%%L. subbaliophaeus `0;F9L9#V" *( FL#= 1 Lactarius K4*%+G&%0 Lactarius K4*%+)&)0 C5;!O and of L. baliophaeus EK$1&$88 ] ;!O; +%%% @0 > @$ > @% > ' containing L. baliophaeus Nigrescentes RESULTS L. subbaliophaeus is a member of this section according to . + ITS rDNA sequence and phylogenetic analyses 9 ;$ F ;00 of the loaned material of Ltenellus and Lkabansus fell into .+9E! @@>Lactarius s. str. !<;!+%%> LPlinthogali clade and constitutes Lactarius s. str! ;+%%> $0+%; ; Lactarius GLactarius afroscrobiculatus'4D01@@+ PlinthogaliPlinthogali L. tenellus $ L. kabansus $ L. miniatescens + L. saponaceus + Lactarius C4G@++ Lactarius miniatescensC4+88+C4+%%+ ;!! ;!!!Russula Lactarius subbaliophaeusC6sp. nov. !<9" C*C*&%8)1& The sequences of Lactarius .$M0 ! Lactarius Etymology? ; L. torminosus * L. wenquanensis from L. baliophaeus

42 IMA FUNGUS Lactarius s. str. in Togo

Table 2.FLactarius baliophaeus and those of L. subbaliophaeus<6/$%+% ARTICLE Features L. baliopheus L. griseogalus L. subbaliophaeus * LG%M8%M@%= LG) L0%M)1= 0'G910*G9 91U4G)4VG 1 140911V091

5 * * * 5_`0_+$G_)A distant (total 28) 5_`0M1A cm U" .9 < . 9

5" /9 7 ; 9 8'$

#".FO0 K / *

Taste C C * * E K E subglobose

~`+%+M+%8M+%@M++1 ~`+%8M+$+M+G)M+11 ~`+%0M+++M+$8 L L L U L0%M11&@M++3 L1%M)1& LG1M1)&)M@ 8M+%3 3 9 9 9 Marginal cells $+M0%&$MGM13 $%MG1&0M83 $GM8$&GM)3 mostly fusiform " "

L 7+%M$1& L$1M0% L$%M)%& GM139 &GM139 GM139

Diagnosis?L 5_ ` 091A XStipeG%M01&&M+G 5 * 0*GContext " 9 Latex C 8'$=taste bitter " Chemical reaction? "

Lactarius subbaliophaeus R .FO0 Basidiospores " = " &M8.5M@&8M7.5M&M&13Q`+%0M1.11M+$8=

.FO0= n ` 81 %1M+3 Type: Togo: Central region? L ;X *09$%M))& (L.R9C%& 0$R++RR(% 0)R$1RRV ++1M+0399 on soil in gallery forest dominated by Uapaca guineensis and 0M8M+%&+M2MG3PleurocystidiaG1M1)&)M@ Afzelia africana +) ^ $%++ Dao Maba MD100 ;OEO 3 – holotype= EV(; M E* X9Pleuropseudocytidia0M5M)3 7E@+8G8$ 5 Marginal cells of lamellae Description: Pileus0%M)1 $GM8$&GM)3 9" 9 "Hymenophoral trama = " Pileipellis 91U4G)4VG $$MG1&GM13 Margin 9Stipitipellis Lamellae "

VOLUME 5 · NO. 1 43 Maba et al. ARTICLE

Fig. 2. 5 Lactarius subbaliophaeus C4+%% A. L B. L C. C D. 7 E.LF.F*`+%3

44 IMA FUNGUS Lactarius s. str. in Togo ARTICLE

AB

G3 $3

C

G3

Fig. 3.FVCLactarius subbaliophaeusC4+%%A–B.*C.E

Clamps DISCUSSION Lactarius subbaliophaeus differs from L. baliophaeus Additional specimen examined: Togo: Central region? L and L. griseogalus in the 9 ' . # 'X %@ +G@R&+RR ( %+ ++0R0$RR V Isoberlinia tomentosa and Uapaca C $)C$%%&Dao Maba MD14;OEO "= ;

VOLUME 5 · NO. 1 45 Maba et al. ARTICLE

A

B C

Fig. 4.*Lactarius subbaliophaeusC4+%%A.EB.LC.5" "*`+%

" ! 4 !;F 4(' easily " that L. subbaliophaeus deviates from L. baliophaeus by \ +$>8%%F

".FO0=;$ U +%%>@@>; R;$L. subbaliophaeus differs L. baliophaeus most from L. griseogalus and is closest to L. baliophaeus ] Lactarius K4*%+G&@@ULactarius spK4*%+G@)@

46 IMA FUNGUS Lactarius s. str. in Togo

Table 3Lactarius;$%%8M$%+G ARTICLE Species Specimens Collector/Date Locality and ecosystem type Lactarius Dao Maba 'XIsoberlinia tomentosa and C4G@++ +)^$%+G Uapaca togoensis%@ +GR)RR(%+ ++0RG&RRV Lactarius Andre De Kesel .RIsoberlinia doka and Uapaca afroscrobiculatus *#+)G0889G$=+)G0+&98@ +)^$%%8 togoensisAAfzelia africana%& 0G@R)GP(% 08)R80PV Atsu Guelly 'XUapaca guinensis Isoberlinia U$$)&2 G+C$%%& doka and Uapaca togoensis%@ +)RG0%P(%+ +GRG$0PV Lactarius atro- Andre De Kesel 'X olivinus *#+)G)809G1 ++^$%%8 %@ +)R0)RR(%+ +$RR0+)RV

*#+)G0@%901 $GC$%%& ! .Uapaca togoensis %8 G)R80P(% 1+R+%1PV '4D01G&G='4D0&+G='4D0&G) +1C$%+% O and +8C$%+% Uapaca guineensis %8 GGR+$GP(% 1$RG8&PV Dao Maba 'X and C4%8 @F$%%& Uapaca guineensis %@ +)R01RR(%+ ++R08RRV Lactarius Dao Maba .RIsoberlinia doka and Uapaca kabansus C4+G$ +@^$%++ togoensis %& 01R$0RR(%% 0&R%&RRV Lactarius Andre De Kesel melanogalus 'X '4D0$@$ ++^$%%8 %@ +)R0)RR(%+ +$R0+)RRV Lactarius Andre De Kesel ;EMilletia thonningii miniatescens *#+)G&$&9@$ +G^$%%8 Lonchocarpus sericeus %& 1))R@$(QR%+ G+@R))VQR Dao Maba .RIsoberlinia doka and Uapaca C4%@ $8C$%%& togoensis%& 0&R1$G1RR(% 01R$@$RRV .RIsoberlinia doka and Uapaca C4+1+ +@^$%++ togoensis%& 0$R$+RR(%% 0)R+&RRV

'X and C40%+ +&^$%+G Uapaca guineensis %@ +)R$&1RR(%+ +GR$+1RRV Lactarius Dao Maba Subbaliophaeus .RUapaca togoensis and Afzelia C4+%% +)^$%++ africana%& 0$R$+RR(% 0)R$$RRV Dao Maba 'XIsoberlinia tomentosa and C4+0 $)C$%%& Uapaca togoensis %@ +GR&RR(%+ ++0R0$RRV Lactarius Andre De Kesel 'XIsoberlinia tomentosa saponaceus *#+1&0+&9+8 ++^$%%8 %@ +G@R&+RR(%+ ++R00$RRV Dao Maba 'XIsoberlinia tomentosa C4G@% +&^$%+G %@ +GR$8RR(%+ ++R1GRRV Lactarius tenellus Andre De Kesel .RUapaca togoensis %& 0GR+01RR *#+)G8@&9)$ $%^$%%8 (% 0)RGG$RRV Atsu Guelly .RIsoberlinia doka and Uapaca U$+0$ G^$%%& togoensis%& 0GR+@RR(%% 0)RG+RRV Dao Maba .RIsoberlinia doka and Uapaca C4+0@=+$)=+1@ +&M+@^$%++ togoensis%& 01R$0RR(%% 0&R%&RRV=%& 0$R$1RR(%% 0)R$0RRV +C4`D. Maba=2U`A. Guelly=G'4D`A. De Kesel.

L. subbaliophaeus forms a terminal sister ; ]K4*%+G%&%0 L torminosus L. wenquanensis L purpureus K4*%+)&)0 Lactarius subbaliophaeus and L. baliophaeus ; belong both to Lactarius sect. Nigrescentes .+ corroborated by 9;+ Lactarius Piperites /;;G 79Uet al+@@&<6/$%+% sequence of L. afroscrobiculatus of Lactarius Piperites ' ; Lactarius

VOLUME 5 · NO. 1 47 Maba et al.

C4G@+ L Piperites &8>'LactariusPiperites " L barbatus and L are still to be collected and acrissimus ' Lactarius C4G@+

ARTICLE C9 ACKNOWLEDGMENTS ; " L tenellus L kabansus L and L saponaceus from Togo and L /!.F angiocarpus] !.F 4A1+8&9+ E ' V" F of L Plinthogali L Plinthogali as 4''4 'A++A8$1)$ E # . .+ and 4.EE]9+80A$9+ L. miniatescens form a clade sister to LNigrescentes L. kabansus and L. tenellus belong to a different +%%>.+FPlinthogali REFERENCES +$ U L. Plinthogali " ' D ( # 5 D97 '" !^ V K Togo from LR V F 7= D DX= # 5 V L F # ; ;'.F;5< ;K*C5D 9 LKDXK$%+%;K(!;V E 9 New Phytologist 186?$&+M$&1 ' 'U^95..$%%$ Carte de Végétation A du TogoL?5*V< 'Lactarius s. str ' F. C ;5 F '' ] ^ ] ] C ( / / 4^ +@@8 E *5'F; LF! *5'F;? Lactarius s. str * et al Nucleic Acids $%%&$%+% Research 25?GG&@MG0%$ V Lactarius atro-olivinus 'D+@&@Recherches sur les forêts denses humides du L. and L. miniatescens Togo; 4K *"!!! *'C4#C*'E4 'E$%+$ VR'Mycorrhiza ; G C in a gallery or savanna 22?+M$@ L. afroscrobiculatus in habitats that harbour *4(#7KCC/$%+%U Uapaca Isoberlinia doka= L. atro-olivinus occurs 4(' and Uapaca Applied Microbiology and Biotechnology 87? @@M guineensis= L.miniatescens Uapaca ! +%& L. tenellus * 4' D9CR ! 5 4^ O ^ / 45 U. togoensis and I. doka as native $%%&E*Nucleic Acids Research 36?$1MG% Ra tendency to U. togoensis. * * 7 <<' / # $%+% L Lactarius saponaceus seems to conserve LactariusBasidiomycota of I. tomentosaLactarius kabansus Taxon 59?$@1M$@) U9]R * * 7 < V K < ' D . $%%& E9F /Russula and lactarius: the dilemma I. doka and U. togoensis of RussulaOchricompactaeFungal Diversity 28?+1M Lactarius subbaliophaeus in savanna 0% and I. tomentosa or 4 D' E D' $%%8 Les Champignons de la chaine de Afzelia africana. l’Atakora#F *"?^ *(* 4 Lactarius s. 4D'UX^;UF($%%$Guide des Champignons str.; Comestibles du Bénin.*"?^*( (Maba et al. $%+G both genera * being collected in the same Lactarius s. str. 4 'EF C'E'4 C4**'C " " C F * E $%+% C9 R O sections including E Chromospermi Piperites Amari Russularia Nigrescentes Environmental Microbiology 12: Plinthogali Pseudofuliginosi $$+@M$$G$ 'Togo 4 C * 'C ; 4 $%%$ 5 (Piperites Nigrescentes and Pseudofuliginosi R  781>Lactarius R'RO? harvested in /' are found in the ecosystems of ; Bois et Forêts des Tropiques 13?1+M)G

48 IMA FUNGUS Lactarius s. str. in Togo

EC*;4+@@G!;F ? ARTICLE R CGlobal Change Molecular Ecology 2?++GM++& Biology 14?+8%GM+8$% 7;$%%1BioEdit<8$1Biological sequence alignment # ; 4 'E 4 C F E 7X D editor for Win95/98/NT/2K/XPUU'?!* <'**4**E*'C$%%8E 79U ^ <' < ^ +@@& The Genus R * ' Lactarius . ( V $ U? !Mycorrhiza 17?0+1M0$& F F<U#*<'L#ED5#$%%1 DD;7$%%&#C'..; ;?4(' Brief Bioinform 9? $&)9$@& ? Philosophical Transactions of the Royal Society of +%+%@GAA%+G !360?+&%1M+&++ DX K 5 D97 ' D ( #7 '" F4(^<'$%+%U !^ V K V F 7= D DX= # 5 V Lactarius gerardii"RussulalesFungal Biology L;F#;'.F;5< ;K 114?$8+M$&G *C $%%1 K(!;V?' 9 ; D L 4 L ( F E ( C D F R New $%++CVE'1?CVE' Phytologist 166?+%)GM+%8& C"5V 4 C" D ' / ^7 +@8& Methuen Handbook of Colour LCMolecular Biology and Evolution 28?$8G+M 5?C $8G@ 5 7; 7 FC $%%8 / #XL4D'<'$%%GF' Mycological Research LactariusRussulalesBasidiomycota++# 111?+%)0M+%80 *Nova Hedwigia 77?$$+M$1+ C45E'D(F<''#$%+G; < ' ( ^ * * $%++ ( .R + subgenera Edules Lactariopsis C ( L ; / ' Mycological RussulopsisMycotaxon 118?008M01G Progress?+%+%%8A++1189%+G9%@G$90 <'/#$%+%Monograph of Lactarius in Tropical C 45 $%+% Le genre Lactarius de la Réserve de Faune Africa..;'$*?( d’AledjoC 4V'*4 *E* C5 ?K 5 < ' * * $%%+ 4 (#7#CDV'D5D97 R ' ! Tropical Mycology / DX K $%%) ;" # 4(' F #.^U''C!F#U1: LF4?'.LPLoS ONE ++M$0/?U'*!L 1+?1@?+%+G8+AX%%%%%1@ /;/CD!;4O.$%%0C #" U^ L #E # ( #R 'C RPrunus africana #X ^* #'L/ F ( #' F 4' $%%& V" VNew Phytologist 161?1+8M1$&

VOLUME 5 · NO. 1 49