全 文 ：North American Truffles in the Tuberaceae : Molecular
and Morphological Perspectives
Bonito Gregory1
?
, Trappe James M. 2 * , Vilgalys Rytas1
(1 Department of Biology, Duke University, Durham, North Carolina 27708 U .S. A . ;
2 Department of Forest Science, Oregon State University, Corvallis, Oregon 97331-5752 U .S. A . )
Abstract: The truffle genus Tuber ( Ascomycota, Pezizales, Tuberaceae) produces underground mushrooms widely sought
as edible fungi . Tuber species are distributed throughout Northern hemisphere forests and form obligate ectomycorrhizal
symbiosis with treeswithin the Pinaceae, Fagaceae, Betulaceae, and Juglandaceae . Of the approximately 100 species of
Tuber worldwide, half are suspected to be endemic to North America . In this study we use multiple genetic loci to assess
patterns of phylogenetic diversitywithin Tuber in order to infer species boundaries and to define morphological and phylo-
geographic species groupings . Sevenmajor cladeswereresolved ( Aestivum, Melanosporum, Rufum, Canaliculatum, Gib-
bosum, Puberulum, and Maculatum) . Two morphologically distinctive species T. asa & T. excavatumwere unresolved .
Thethreemost specious clades of Tuber ( Rufum, Puberulum, Maculatum) are distributed across Europe, Asia, and
North America and are comprised mainly of non-commercial species .
Key words: Taxonomy; Tuber; Phylogenetics
CLC number : S 646 Document Code : A Article ID: 0253 - 2700 ( 2009) Suppl.ⅩⅥ- 039 - 13
Introduction
The‘true truffles’are Ascomycete fungi belong-
ing to the genus Tuber and are distributed throughout
the northern hemisphere . There are estimated to be
around 100 species of Tuber worldwide, half of which
appear to be endemic to North America . Tuber , be-
longs within the Tuberaceae, a family that includes
northern hermisphere genera ( Tuber , Choiromyces) and
southern hemispheregenera ( Reddellomyces, Dinglyea,
Labyrinthomyces) (O′Donnell et al. , 1997) .
Tuber species formectomycorrhizal (EM) symbio-
sis with many important timber and nut tree species,
including those belonging to the Pinaceae, Fagaceae,
Salicaceae, Betulaceae, and Juglandaceae . These hy-
pogeous fungi have coevolvedwithmammals, which are
known to be important agents in the dispersal of truffle
spores . Additionally, many European and a few North
American Tuber species areprized edibles andhavethe
potential to be lucrative agricultural crops .
The first valid description of thegenus Tuber was
by F . H . Wiggers (1780 ) , according to the current
International Codeof Botanical Nomenclature (Greuter,
1999) . Wiggers recorded a single species from north
Germany, named it T . gulosorum and described it as
( translated from Latin) :“ a subglobose fungus filled
with tasty flesh .”While this description isof little tax-
onomic value, it meets the requirements of the Code
(Trappe, 2001) . The first useable descriptionsof spe-
cies of Tuber were by Vittadini ( 1831 ) , whose collec-
tions representing most of his species are preserved in
good condition at the University of Turin herbarium in
Italy .
The taxonomic literatureon North American Tuber
spp . began with H .W . Harkness (1899) (Fig. 1) and
was expanded by Helen Gilkey ( 1916 , 1939 , 1947 ,
1954) . Other useful contributions include thoseof But-
ters ( 1903 ) , Trappe and Guzmán ( 1971 ) , Trappe
(1975 , 1979 ) , Uecker and Burdsall (1977) , Cázares
et al . ( 1992 ) , Trappe et al . ( 1996 ) , Colgan and
Trappe ( 1997 ) , Trappe and Castellano ( 2000 ) ,
Trappe and Cázares ( 2000, 2006 ) and Frank et al .
(2006 ) . Fortunately, unlike many European species,
all described North American species are represented by
type collections . Currently, are recognized (Table 2) .
云 南 植 物 研 究 2009 , Suppl . ⅩⅥ : 39～51
Acta Botanica Yunnanica
? ?Author for correspondences; E-mail : gmb2 @duke. edu; trappej@onid. orst. edu
Fig . 1 Pioneers of North America Truffle Taxonomy
A . Taxonomy of North American truffles began with Dr . Harvey Harkness (1821 - 1901 ) . B . Dr . Helen Gilkey ( 1886 - 1972) de-
scribed and illustrated many new species of North American truffles . C . Dr . J im Trappe ( 1933-present) has described and revised
truffle taxonomy and systematics throughout his career
At present, about250 names havebeen applied to
species and varieties in thegenus Tuber . Many species
have been namedmore than once by different authors .
By eliminating probablesynonyms, we estimate for Eu-
rope about 33 of species of Tuber are recognized ( Table
1) (Berkeley and Broome, 1846; Ceruti et al. , 2003;
Montecchi and Sarasini , 2000; Paolocci et al. , 2004)
and 35 species of Tuber in North America . For Asia at
least 11 species of Tuber have been validly described
(Table 3 ) , and an unknown and likely large number
are still undescribed (Cooke and Massee, 1892; Wang
et al. , 1998; Wang and He, 2002; He et al. , 2004 ,
Zhang et al. , 2005; Hu and Wang, 2005; Chen et al. ,
2005; Chen and Liu, 2007; Jeandroz et al. , 2008) . No
endemic Tuber species are known from the southern
hemisphere . Speciesof Tuber described fromAustrala-
sia andSouth Americahavein all casesbeen associated
with trees introduced fromthe northern hemisphere and
probably came as hitchhikerson roots of imported seed-
lings (Trappe and Cázares, 2000) .
The main goal of this research is to define mor-
phological and phylogeographic groupings within the
Tuberaceae, particularly within species complexes and
widely distributed taxa (e.g . T. rufumcomplex, T. m-
aculatum complex) . In addition, using the resulting
phylogenetic framework, we aimto place sequences of
Table 1 Tuber species described from Europe Species with
commercial interest are labeled with an asterix ( * )
Tuber species in Europe
* T ?. aestivum ( = T. uncinatum) Vittadini
T J. asa Tulasne & Tulasne
T J. belionaeQuelet
* T ?. borchii Vittadini
* T ?. brumaleVittadini
T J. dryophilumTulasne & Tulasne
T J. excavatumVittadini
T J. ferrugineum Vittadini
T J. foetidumVittadini
T J. fulgens Quelet
T J. lutescens Lazaro & Ibiza
* T ?. macrosporumVittadini
T J. maculatumVittadini
* T ?. magnatumPico
T J. malenconi Donadini , Riousset, Tiousset & Chevalier
* T ?. melanosporumVittadini
T J. mesentericumVittadini
T J. microsporumVittadini
T J. michailowskoanumBucholtz
T J. murinum Hesse
T J. multimaculatum Parlade, Trappe & Alvarez
T J. nitidum Vittadini
T J. oligospermum (Tulasne & Tulasne) Trappe
T J. pallidumLazaro & Ibiza
T J. panniferumTulasne & Tulasne
T J. puberulumBerkeley & Broome
T J. queietianumFerry de la Bellone
T J. rapoeodorumTulasne & Tulasne
T J. regianumMontecchi & Lazzair
T J. rufumPico
T J. scruposumHesse
T J. sinuosum Lazaro & Ibiza
T J. verii Pacioni & Lalli
04 云 南 植 物 研 究 增刊ⅩⅥ
Tuber from ectomycorrhizal root tips and unidentified
fruitbodies into this phylogenetic framework .
Fig . 2 Morphological Variation of Tuber
Truffles vary in size along a continuum from very small to very large, as
exemplified by a collection of and unidentified Tuber ( A ) and T . gib-
bosum, respectivly (B) . Tuber species are characterized by a pattern of
sterile and fertileveins filling thegleba, which darken as thefertile tissue
matures and vary in color by species as shownwith T. canaliculatum (C)
and T. lyonii (D) . Other gross morphological features help to distinguish
Tuber species, such as the texture of the peridium . The peridium may
bear largewarts as in T. aestivum (E ) , or it could be rough , scaley, pu-
bescent, or glabrous as in T. oregonense . nom . prov . ( F ) . Microscopic
characters of the asci and spores arenecessary for distinguishing species of
Tuber . Asci may be globose or sub-globose as shown for unidentified Tu-
ber (G ) or more flask-shaped and bearing a stemas in T. lyonii ( H) . Fi-
nally, spores may be alveolate- reticulateor spiny, illustrated by T. macu-
latum ( I ) and T. melanosporum ( J ) , respectively
Table 2 Tuber species described fromNorth America Species
with commercial interest are labeled with an asterix ( * )
Tuber species in North America
T J. anniae Colgan & Trappe
T J. argenteumGilkey
T J. besseyi Gilkey
T J. bellisporumBonito & Trappe nom . prov .
T J. californicumHarkness
* T ?. canaliculatumGilkey
T J. candidumHarkness
T J. castellanoi Bonito & Trappe nom . prov .
T J. citrinumHarkness
T J. clarei Gilkey
T J. gardneri Gilkey
* T ?. gibbosum= T Y. giganteumGilkey
T J. guzmanii Cazarez & Trappe
T J. harknessii Gilkey
T J. irradians Gilkey
T J. levissimumGilkey
T J. linsdalei Gilkey
T J. longisporumGilkey
T J. luomai Trappe nom . prov .
* T ?. lyonii Butters
T J. monticola Harkness
T J. murinum Hesse
* T ?. oregonenseTrappe & Bonito nom . prov .
T J. pacificumTrappe & Castellano
T J. phleboderma (Gilkey) Trappe comb . nov .
T J. quercicola Frank, Southworth, & Trappe
T J. rapaeodorum Tulasne & Tulasne
T J. regimontanumGuevara, Bonito & Rodríguez
T J. separans Gilkey
T J. shearii Harkness
T J. sphaerosporumGilkey
T J. spinoreticulatumUecker et Burdsall
* T ?. texense Heimsch
T J. unicolor Gilkey
T J. whetstonensis Frank, Southworth , & Trappe
Table 3 Tuber species described fromAsia Species with
commercial interest are labeled with an asterix ( * )
Tuber species in Asia
T ?. formosanumHu
T ?. furfuraceumHu & Wang
T ?. huidongenseWang & He
* T 7. indicumCooke & Massee
T ?. latisporumChen & Liu
T ?. liatongense Liu
T ?. liui Xu
T ?. pseudoexcavatumWang, Moreno, Riousset, Manjón & Riousset
T ?. taiyunense Wang
T ?. umbilicatumChen & Liu
T ?. zhongdianense Wang
Methods
Morphological : Tuber spp . formstereothecia with
a distinct, simple to layered peridiumenclosingagleba
of fertile tissue marbled with sterile, hypha-stuffed
14增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .
veins that tend to open through the peridium . The asci
are randomly embedded in the fertile tissue and within
a specimen may contain from one to 4 ( - 6) spores .
The followingcharacter states haveprovenuseful indif-
ferentiating species, in general order of importance .
Spores spiny, spiny with a reticulum of low lines, or
reticulate with tall reticular walls; ellipsoid to subglo-
boseor globose; spores in 1- or 2-spored asci up to45 ,
55 , 65 or 75 μm long . Asci astipitate, with a short
stipe, or a longstipewith a forked base; thin-walledor
withwalls up to 1 , 2 or 4μmthick . Ascomatal surface
smooth, pubescent with outgrowing hyphae, pubescent
with tapered cystidia, scabrous, verrucose with round-
ed warts, verrucose with angular warts, white, gray,
yellow, olive, brown, reddish brown, dark brown or
black . Peridiumasingle, undifferentiated layer or with
two or more layers differentiated by size, shape, wall
thickness, or pigmentation of the cells; greatly inflated
cells absent or present in one or more layers . Glebal
cells similar to those of inner peridium or differentiated
by cell diameter or presence of inflated cells . Species
can usually be differentiated by the large number of
combinations of thesevarious characters, but complexes
of closely related species can be difficult to separate
without supporting molecular data (Mello et al. , 2000;
Halász et al. , 2005; Frank et al. , 2006) .
Molecular: DNA was extracted with 24∶1 chloro-
form: isoamyl alcohol and precipitated in isopropanol .
Both the internal transcribed spacer region ( ITS1 ,
5 .8S, and ITS2 ) and three divergent domains ( D1 ,
D2 , D3 ) of the ribosomal large subunit ( LSU) locus
were amplified using the universal fungal primer set
ITS5-LR5 (Bertini et al. , 1999; Vilgalys and Hester,
1990) . Other loci fromtheseextractionswere PCR am-
plified, including the ribosomal small subunit (SSU ) ,
elongation factor 1 alpha ( EF1α) , the second subunit
of RNA polymerase ( RPB2) . PCR conditions and the
handling of PCR products were as described in Healy
et al . ( 2009 ) . Sanger sequencing was performed on
an ABI3700 (Applied Biosystems, Foster City, CA )
using Big Dye chemistry version 3 .1 (Applied Biosys-
tems, Foster City, CA ) in both directions . DNA se-
quences were assembled and manually edited using Se-
quencher 4 .0 ( Gene Codes, Ann Arbor, MI ) and
queried against the NCBI public database GenBank
( http:??blast.ncbi .nlm.nih.gov?Blast. cgi ) with the
BLASTN algorithm to compare with other sequences
and to verify that sequences were of the target group .
Sequences were aligned manually using MacClade
( Maddison and Maddison, 2002 ) . Ambiguously
aligned regions were excluded from the analyses . Phy-
logenetic analyses were conducted using maximum par-
simony and maximum liklihood methods . Parsimony
analyses were carried out using a heuristic search in
PAUP 4.0b10 with 1000 random addition sequences
and 5000 bootstrap replicates (Swofford, 2001) . Two
independent maximum likelihood analyses based on a
general-time-reversible 6-parameter model of evolution
were run using the software program GARLI and in-
cluded 100 bootstrap replications (Zwickl , 2006) .
Results
Molecular analyses of LSU and SSU data support
the conclusions of O′Donnell et al . (1997) that there
aredistinct northern hemisphere lineages in theTuber-
aceae, and distinct southern hemisphere lineages
( Fig. 3 ) . Four sourthern hemisphere clades were re-
solved including ( Dingleya, Labyrinthomyces, Red-
dellomyces, and Leppia nom . prov .) ( Trappe and
Claridge, 2005 ) . Resolved and supported Northern
hemisphere clades include Choiromyces and Tuber,
Choiromyces is representedby C. aveolatus and C. ve-
nosus, while Tuber is much more speciose .
Multigene analyses of LSU , EF1a, 5 . 8S, and
RPB2 loci support 7 clades ( Aestivum, Melanospo-
rum, Rufum, Canaliculatum, Gibossum, Puberulum,
Maculatum) within Tuber ( Fig. 4 ) . Tuber asa and
Tuber excavatumwere not well resolved in these analy-
ses .Themost basal clade (Aestivum) was composedof
strictly European taxa ( T. mesentericum, T . aestivum,
T. magnatum) . The Melanosporum (Fig. 5 ) and Ru-
fum ( Fig. 6 ) clades were reprentated by European,
Asian, and North American taxa, and ( aside from
T. pseudoexcavatum) are composedof spiney and spin-
ey-reticulated spored species . In addition, we found
evidence from sequences derived from fruitbody and
24 云 南 植 物 研 究 增刊ⅩⅥ
Fig . 3 Phylogeny of Tuberaceae
The Tuberaceae is represented by four Southern Hemisphere genera ( Dinglyea , Reddelomyces, Labyrinthomyces, and Leppia
nom . prov .) and two NorthernHemispheregenera ( Tuber, Choiromyces) . This phylogenetic analysis is based on the small sub-
unit ( SSU) and large subunit (LSU) rDNA sequence data, and was analyzed using unweighted maximum parsimony
34增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .
Fig . 4 Phylogeny of Tuber
Seven clades of Tuber were resolvedwith unweighted parsimony analyze based on multiple loci ( rDNA LSU , 5 . 8S rD-
NA , RNA polymerase II , and elongation factor alpha 1 ) . These clades are the Aestivum, Melanosporum, Rufum,
Canaliculatum, Gibbosum, Maculatum, Puberulum ( Borchii) groups . The European species Tuber excavatum and
T. asa werenot resolved into any of these clades in this analysis . Nodes with significant support ( > 70 % parsimony
bootstrap) are signified by thickened branches
44 云 南 植 物 研 究 增刊ⅩⅥ
Fig . 5 The Melanosporum Clade
Representatives of the Melanosporum clade in North America include Tuber regimontanum, T. melanosporum, and T. indicum . Tuber mela-
nosporumhas been introduced in efforts to cultivate this species asa cash crop . It is most likely that T . indicum (clade II ) was introduced into
North America through forestry projects early in the century ( e. g . fruitbody collection) and more recently ( accidentally) through T. mela-
nosporum cultivation efforts . These three species are similar in morphology, though T. regimontanumhas larger spores than T. melanosporum
and T . indicum and short reticulations connecting the spines of its spores . Results are based on parsimony analysis of the ITS rDNA region .
Taxa names are followed by collection ( or Genbank accession) numbers, followed by point of origin ( when known)
mycorrhizas suggesting that the Asian species T. in-
dicumhas been introduced into the United States . Two
species were resoloved in the Canaliculatum clade;
T. macrosporum ( Europe ) and T. canaliculatum
(North America) , while the Gibbosumclade was com-
posed of soley of species restricted to thePacific North-
west regionof North America ( Fig. 7) . The Puberulum
(Fig. 8 ) and Maculatum ( Fig. 9 ) clades were repre-
sented by taxafromAsia, Europe, andNorth America,
and together with the Rufumcladeincludes themajori-
ty of Tuber species . They present a taxonomic chal-
lenge to mycologists, owing to a lack of apparent mor-
phological characters for distinguishingspecies fromone
another .
Conclusions
Europe appears to have the most phylogenetic di-
versity of Tuber, particularly represented by basal lin-
eages, suggesting that this continent may be the point
of origin for the genus . However Tuber diversity in
Asia is still not well known and the discovery and in-
clusion of new taxa in future analyses could change in-
ferences about the origin and diversification of Tuber .
Further work should focusonmolecular analysisof
type specimen ( when possible) and attempt a more
comprehensive coverage of the species diversity within
the family Tuberaceae . In addition, it is still undeter-
mined whether the genus Paradoxa and Loculotuber
fall within the Tuberaceae, and if so, whether these
genera aremono- , para- , or polyphyletic .
54增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .
Fig . 6 The Rufum Clade
NorthAmerica is well represented by spiney-spored members of the Rufum clade . As far as we are aware, T . rufum Vittadini
senso stricto does not exist in Asia or North America . Results arebased on parsimony analysis of the ITS rDNA region . Nodes
with significant support ( > 70 % parsimony bootstrap) are signified by thickened branches . Taxa names are followed by collec-
tion (or Genbank accession) numbers, followed by point of origin ( when known) . Names in quotations are from collection labels
and are most likely incorrect
64 云 南 植 物 研 究 增刊ⅩⅥ
Fig . 7 The Gibbosum Clade
To thebest of our knowledgemembers of the Gibbosum clade areonly represented in thePacific Northwest region of NorthAmeri-
ca . The four species include T. gibbosum, T. bellisporum prov . nom ., T . castellanoi prov . nom ., and T. oregonense prov .
nom . . Results are based on maximum liklihood analysis of the ITS and LSU rDNA regions . Nodes with significant support ( >
70 % maximum liklihood bootstrap) are signified by thickened branches . Taxa names are followed by collection numbers
74增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .
Fig . 8 The Puberulum Clade
Many undescribed and unidentified species ( resulting from mycorrhizal molecular studies) belong to the Puberulum
clade . Morphologically, thisgroup tends to haveglobose tosub-globosespores, and fruitbodies that arepale in color and
small in size . Species in this group are found in Europe, North America, and Asia . Results are based on parsimony
analysis of the ITS rDNA region . Nodes with significant support ( > 70 % parsimony bootstrap) are signified by thickened
branches . Taxa names are followed by collection ( or Genbank accession) numbers, followed by point of origin ( when
known) . Names in quotations are from collection labels and are most likely incorrect
84 云 南 植 物 研 究 增刊ⅩⅥ
Fig . 9 The Maculatum Clade
Many undescribed and unidentified species ( resulting from mycorrhizal molecular studies) belong to the Maculatum
clade . Morphologically, this group tends to have sub-globose to elliptical spores, and fruitbodies that are pale in color
and small in size . Species in thisgroup are found in Europe, North America, and Asia . Results arebased on parsimony
analysis of the ITS rDNA region . Nodes with significant support ( > 70 % parsimony bootstrap) are signified by thickened
branches . Taxa names are followed by collection ( or Genbank accession) numbers, followed by point of origin ( when
known) . Names in quotations are from collection labels and are most likely incorrect
94增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .
Acknowledgements : Thanks to Tim James, Jeri Parrent, Heath
O′Brien, Jason Jackson, Bernadette O′Reilly, Hannah Reynol-
ds , Paul Manos, Joey Spatafora, Kentaro Hosaka, Conrad
Schoch, and Doni McKay for providing research guidance .
Thanks to Andre Gryganski , Rosanne Healy, Matt Trappe, Matt
Smith, Gonzalo Guevara, Peter Kennedy, Efren Cazares,
Michael Castellano, Alassandra Zambonelli , and Lisa Bukovnik
for providingcollectionmaterial , photos, technical and labassis-
tance . This research was supported by NSF award # 0641297
‘REVSYS: Phylogenetic and Revisionary Systematics of North
American Truffles ( Tuber , Ascomycota)’.
References:
Berk ?eley MJ , Broome EC . 1846 . Notices of British Hypogeous Fungi
[ J ] . The Annals and Magazineof Natural History, 18 : 73—82
Bert ?ini L , Amicucci A , Agostini D et al. , 1999 . A new pair of primers
designed for amplification of the ITS region in Tuber species [ J ] .
FEMS Microbiology Letters, 173: 239—245
Butt ?ers FK , 1903 . A Minnesotaspecies of Tuber [ J ] . Botanical Gazette,
35 : 427—431
Caza ?res EJ , García J , Castillo et al. , 1992 . Hypogeous fungi from north-
ern Mexico [ J ] . Mycologia, 84 : 341—359
Ceru ?ti AA, Fontana, Nosenzo C , 2003 . Le Specie Europee del Genere
Tuber—Una Revisione Storica . Museo Regionale di Scienze Natura-
li , Torino, Italy .
Chen ?J , Liu PG, WangY , 2005 . Tuber umbilicatum, anew speciesfrom
China, with a key to the spinose-reticulate spored Tuber species
[ J ] . Mycotaxon, 94 : 1—6
Chen >J , Liu PG, 2007 . Tuber latisporum sp . nov . and related taxa,
based on morphology and DNA sequence data [ J ] . Mycologia, 99 :
475—481
Colg ?an III W, Trappe JM , 1997 . NATS truffle and truffle- like fungi 7 .
Tuber anniae sp . nov . ( Ascomycota) [ J ] . Mycotaxon, 64 : 437—
441
Cook e MC , Massee G, 1892 . Himalayan truffles [ J ] . Grevillea, 20 : 67
Cord ?a ACI , 1842 . Icones fungorum hucusque cognitorum . Vol . 5 . Pra-
gue .
Fran ?k JL , Southworth D, Trappe JM, 2006 . NATS truffle and truffle-like
fungi 13: Tuber quercicola and T. whetstonense, newspecies fromOre-
gon, and T. candidumredescribed [ J ] . Mycotaxon, 95 : 229—240
Gard #es M, Bruns TD, 1996 . Community structureof ectomycorrhizal fun-
gi in a Pinus muricata forest: Above- and below-ground views [ J ] .
Canadian J ournal of Botany, 74 : 1572—1583
Gilk ?ey HM, 1916 . A revision of theTuberales of California [ J ] . Univer-
sity of California Publications in Botany, 6 : 275—356
Gilk ?ey HM , 1939 . Tuberales of North America [ J ] . Oregon State Mono-
graphs in Botany, 1 : 1—63
Gilk ?ey HM , 1947 . New or otherwise noteworthy species of Tuberaceae
[ J ] . Mycologia, 39 : 441—452
Gilk ?ey HM , 1954 . Tuberales [ J ] . North American Flora Ii , 1 : 1—36
Greu #ter WJ , Mcneill FR , Barrie HM et al. , 1999 . International Codeof
Botanical Nomenclature ( St . Louis Code) . Sixteenth International
Botanical Congress . St Louis, Missouri , J uly-August 1999 . Con-
gress . RegnumVegetabile 138 . Koeltz Scientific Books, K?nigstein
HeX ?Y , Li HM , Wang Y , 2004 . Tuber zhongdianense sp . nov . from
China [ J ] . Mycotaxon, 90 : 213—216
Hu H ?T , 1992 . Tuber formosanum sp . nov . and its mycorrhizal associa-
tions [ J ] . Quart J Exp Forest, 6 : 79—86
Hu H ?T , Wang Y , 2005 . Tuber furfuraceumsp . nov . fromTaiwan [ J ] .
Mycotaxon, 93 : 155—157
Halá ?szKZ, Bratek D . Szeg, Rudnóy S et al. , 2005 . Tests of species
concepts of the small , white, European group of Tuber species based
on morphology and rDNA sequences with special reference to Tuber
rapaeodorum [ J ] . Mycological Progress, 4 : 291—298
Hark ?ness HW, 1899 . California hypogaeous Fungi [ J ] . Proceedings of
the California Academy of Science III , 1 : 241—292
Heal ?y RA , Bonito G, Guevara G, 2009 . The truffle genus Pachyphloeus
in the U. S . and Mexico: phylogenetic analysis and a new species
[ J ] . Mycotaxon, 107: 61—71
J ean ?droz S, Murat C, Wang Y et al. , 2008 . Molecular phylogeny and
historical biogeography of the genus Tuber [ J ] . J ournal of Biogeog-
raphy, 35 : 815—829
MaddisonDR , Maddison WP, 2002 . MacClade: Analysis of Phylogeny
andCharacter Evolution
Mello A , Vizzini A , Longato S et al., 2000 . Tuber borchii versus T. m-
aculatum: neotype studies and DNA analyses [ J ] . Mycologia, 92 :
326—331
Montecchi A , Sarasini M , 2000 . Funghi ipogei d′Europa . Trento, Italy:
Associazione Micologica Bresadola
O′D ?onnell K , Cigelnik E , Weber N et al. , 1997 . Phylogenetic relation-
ships among ascomycetous truffles and the true and false morels in-
ferred from 18s and 28s ribosomal DNA sequence analysis [ J ] . My-
cologia, 89 (1 ) : 48—65
Paolocci FA , Rubini R , Riccioni C et al. , 2004 . Tuber aestivum and
Tuber uncinatum: two morphotypesor twospecies [ J ] . FEMS Micro-
biology Letters, 235 ( 1) : 109—115
Swof ?ford DL , 2001 . PAUP* . Phylogenetic Analysis Using Parsimony ( *
and Other Methods ) . Version 4 . Sinauer Associates, Sunderland,
Massachusetts
Trappe JM , 1975 . Generic synonyms in the Tuberales [ J ] . Mycotaxon,
2 : 109—122
Trap ?pe JM , 1979 . The orders, families, and genera of hypogeous Asco-
mycotina ( truffles and their relatives ) [ J ] . Mycotaxon, 9 ( 1 ) :
297—340
Trap ?pe JM, 2001 . Taxonomic and nomenclatural problems in the genus
Tuber . In Science et Culturede laTruffe: ActesduVth Congrès In-
ternational, Aix-en-Provence, 4 au 6 Mars . 1999 . Fédération
Fran?aise des Trufficulteurs, Paris, 112—114
Trap ?pe JM, Castellano ML , 2000 . New hypogeous Ascomycota and Ba-
sidiomycota covered by the Northwest Forest Plan [ J ] . Mycotaxon,
75 : 153—180
Trappe JM , Cázares E , 2000 . Tuber maculatum around the world [ J ] .
Bulletin Semestriel de la Fédération des Associations Mycologiques
05 云 南 植 物 研 究 增刊ⅩⅥ
Méditerranénnes n. s. , 18 : 107—112
Trap ?pe JM , Cázares E , 2006 . Tuber guzmanii , a new truffle fromsouth-
ern Mexico [ J ] . International J ournal of Medicinal Mushrooms . ( in
press)
Trap ?pe JM, Claridge AW, 2005 . Hypogeous fungi : evolution of repro-
ductive and dispersal strategies through interactions with animals and
mycorrhizal plants [ A ] . In: Dighton et al. ( eds .) , The Fungal
Community— Its Organization and Role in the Ecosystem [ M ] . 3rd
Ed . Boca Raton: Taylor & Francis, 613—623
Trap ?pe JM , Claridge A , 2006 . Leppia: A missing link betweenNorthern
and Southern hemisphere truffles ? [ J ] . Inoculum: Supplement to
Mycologia, 57 (1) : 14—15
Trap ?pe JM , Guzmán G, 1971 . Notes on somehypogeousfungi fromMexi-
co [ J ] . Mycologia, 63 : 317—332
Trap ?pe JM , Jumpponen A , Cázares E , 1996 . NATS truffle and truffle-
like fungi 5: Tuber lyonii ( = T. texense) , with a key to the spiny-
spored Tuber species groups [ J ] . Mycotaxon, 60 : 365—372
Ueck +er FA , Burdsall HH J r, 1977 . Tuber spinoreticularum, anew truffle
fromMaryland [ J ] . Mycologia, 69 : 626—630
Vilg ?alys R, Hester M, 1990 . Rapid genetic identification and mapping of
enzymatically amplified ribosomal DNA fromseveral speciesof Crypto-
coccus [ J ] . J ournal of Bacteriology, 172: 4238—4246
Vitt ?adini C , 1831 . Monographia Tuberaceum . Felicis Rusconi , Milan
WangY , He XY , 2002 . Tuber huidongense sp . nov . from China [ J ] .
Mycotaxon, 83 : 191—194
Wang ?Y , Moreno G, Riousset LJ et al. , 1998 . Tuber pseudoexcavatum
sp . nov . a new species from China commercialised in Spain,
France and I taly with additional comments on Chinese truffles [ J ] .
Cryptogamie, 19 : 113—120
Wigg ?ers FH , 1780 . Primitiae Florae Holsaticae . M . F . Bartsch, Kiel
Xu A ?S, 1999 . Notes on the genus of Tuber from Tibet [ J ] . Mycosys-
tema, 18 : 361—365
Zhan ?g L , Yang ZL , Song DS, 2005 . A phylogenetic study of commercial
Chinesetruffles and their allies: Taxonomic implications [ J ] . FEMS
Microbiology Letters, 245 : 85—92
Zwic ?kl D, 2006 . Genetic algorithmapproachesfor the phylogenetic analy-
sis of large biological sequence datasetsunder themaximum likelihood
criterion . Ph . D . dissertation . University of Texas . Austin
15增刊ⅩⅥ Bonito Gregory et al. : North American Truffles in theTuberaceae: Molecular and Morphological . . .