American Journal of Botany 86(9): 1325–1345. 1999.
SYSTEMATICS
CLADISTIC
AMARYLLIDACEAE BASED ON
ANALYSIS OF PLASTID RBCL AND TRNL-F
OF
SEQUENCE DATA1
ALAN W. MEEROW,2,3 MICHAEL F. FAY,4 CHARLES L. GUY,5
QIN-BAO LI,5 FARIDAH Q. ZAMAN,4 AND MARK W. CHASE4
3
University of Florida, Fort Lauderdale Research and Education Center, 3205 College Avenue,
Fort Lauderdale, Florida 33314;
4Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3DS, UK;
5University of Florida, Department of Environmental Horticulture, 1545 Fifield Hall, Gainesville, Florida 32611
Cladistic analyses of plastid DNA sequences rbcL and trnL-F are presented separately and combined for 48 genera of
Amaryllidaceae and 29 genera of related asparagalean families. The combined analysis is the most highly resolved of the
three and provides good support for the monophyly of Amaryllidaceae and indicates Agapanthaceae as its sister family.
Alliaceae are in turn sister to the Amaryllidaceae/Agapanthaceae clade. The origins of the family appear to be western
Gondwanaland (Africa), and infrafamilial relationships are resolved along biogeographic lines. Tribe Amaryllideae, primarily
South African, is sister to the rest of Amaryllidaceae; this tribe is supported by numerous morphological synapomorphies
as well. The remaining two African tribes of the family, Haemantheae and Cyrtantheae, are well supported, but their position
relative to the Australasian Calostemmateae and a large clade comprising the Eurasian and American genera, is not yet
clear. The Eurasian and American elements of the family are each monophyletic sister clades. Internal resolution of the
Eurasian clade only partially supports currently accepted tribal concepts, and few conclusions can be drawn on the relationships of the genera based on these data. A monophyletic Lycorideae (Central and East Asian) is weakly supported.
Galanthus and Leucojum (Galantheae pro parte) are supported as sister genera by the bootstrap. The American clade shows
a higher degree of internal resolution. Hippeastreae (minus Griffinia and Worsleya) are well supported, and Zephyranthinae
are resolved as a distinct subtribe. An Andean clade marked by a chromosome number of 2n 5 46 (and derivatives thereof)
is resolved with weak support. The plastid DNA phylogenies are discussed in the context of biogeography and character
evolution in the family.
Key words:
Amaryllidaceae; cladistic analysis; molecular systematics; moncotyledons; phylogeny; plastid DNA.
The Amaryllidaceae J. St.-Hil., a cosmopolitan (predominantly pantropical) family of petaloid monocots,
represent one of the elements of the Linnaean Hexandria
monogynia (Linnaeus, 1753), the 51 genera of which
have been variously classified since as liliaceous or amaryllidaceous. This basic dichotomy represents the generally uncertain phylogenetic placement of many petaloid
monocots until the past two decades. Seven of the 51
genera that Linneaus placed in Hexandria monogynia
have since been included within a common taxonomic
unit, as section Narcissi (Adanson, 1763; de Jussieu,
1789), family Amaryllideae (Jaume-St.-Hilaire, 1805;
Brown, 1810), order Amaryllidaceae (Lindley, 1836;
Herbert, 1837), tribe Amarylleae (Bentham and Hooker,
1883), suborder Amarylleae (Baker, 1888), subfamily
Amaryllidoideae (Pax, 1888); family Amaryllidaceae
1 Manuscript received 10 August 1998; revision accepted 14 January
1999.
Significant portions of this work were completed during the senior
author’s sabbatical leave at the Royal Botanic Gardens, Kew. Some of
the sequences were generated at the DNA Sequencing Core of the Interdisciplinary Center for Biotechnology Research at the University of
Florida. Financial support was provided by NSF grants DEB-968787
and IBN-9317450 to AWM and CLG, a Research Enhancement Award
from the Institute of Food and Agricultural Sciences of the University
of Florida, and the Royal Botanic Gardens, Kew. The authors thank
David L. Swofford for allowing the use of experimental versions of his
PAUP* software. Florida Agricultural Experiment Station Journal Series
Number R-03602.
2 Author for correspondence.
(Hutchinson, 1934, 1959), and subfamily Amarylloideae
(Traub, 1963). Brown (1810) was the first to propose that
the genera with superior ovaries be excluded from Amaryllidaceae, a restriction followed faithfully until Hutchinson (1934). Herbert (1837) recognized that the Taccaceae were not allied to Amaryllidaceae, and Pax (1888)
formally removed Velloziaceae as part of the family (Herbert’s suborder Xerophyteae). Hutchinson’s (1934, 1959)
classification was the first radical recircumscription of
Amaryllidaceae since Brown (1810). In defining the unifying character of the family to be ‘‘an umbellate inflorescence subtended by an involucre of one or more spathaceous bracts,’’ he segregated Agavaceae, Hypoxidaceae, and Alstroemeriaceae and added tribes Agapantheae, Allieae, and Gilliesieae (Alliaceae). Takhtajan
(1969) recognized Amaryllidaceae in the narrowest
sense, and maintained a distinct Alliaceae. Cronquist
(1988) and Thorne (1976) included Amaryllidaceae within broad concepts of Liliaceae.
Concepts of familial and ordinal limits of the monocotyledons were radically challenged by Huber (1969),
who emphasized less conspicuous characters, particularly
embryological characters, over gross floral or vegetative
morphology. Huber’s work highlighted the heterogeneity
present in many traditional monocot families, especially
Liliaceae. Much of this work was refined and placed into
phylogenetic context by Dahlgren and coworkers (Dahlgren and Clifford, 1982; Dahlgren and Rasmussen, 1983;
1325
1326
AMERICAN JOURNAL
Dahlgren, Clifford, and Yeo, 1985). In Dahlgren, Clifford, and Yeo’s (1985) synthesis, Amaryllidaceae and Alliaceae are both recognized as members of the order Asparagales, an order of 31 families that have evolved many
traits in parallel with Liliales. One of the most important
and consistent characters separating these two orders is
the presence of phytomelan in the seed coat of Asparagales (Huber, 1969). To date, phylogenetic analyses of
the monocotyledons, based on both morphological and
gene sequence matrices, have supported this classification
with some amendment (Duvall et al., 1993; Stevenson
and Loconte, 1995; Chase et al., 1995a, b), but the precise relationship of Amaryllidaceae to other Asparagales
remained elusive until Fay and Chase (1996) used molecular data to argue that Amaryllidaceae, Agapanthaceae, and Alliaceae form a monophyletic group and that
together they are related most closely to Hyacinthaceae
s.s. and the resurrected family Themidaceae (the former
tribe Brodiaeeae of Alliaceae).
Despite a lack of consensus on generic limits and tribal
delimitations within the Amaryllidaceae, cladistic analysis has only rarely been applied to problems in the family,
such as by Nordal and Duncan (1984) for Haemanthus
and Scadoxus, two closely related, baccate-fruited African genera, Meerow (1987a, 1989) for Eucrosia and Eucharis and Caliphruria, respectively, and Snijman (1994)
and Snijman and Linder (1996) for various taxa of tribe
Amaryllideae. Applying phylogenetic studies for the entire family is difficult due to homoplasy for many conspicuous characters within this highly canalized group
(Meerow, 1987a, 1989, 1995). This led Meerow (1995)
to conclude that ‘‘future reconstruction attempts will
greatly benefit from the inclusion of molecular data.’’
The four most recent infrafamilial classifications (Table
1) of Amaryllidaceae are those of Traub (1963), Dahlgren, Clifford, and Yeo (1985), Müller-Doblies and Müller-Doblies (1996), and Meerow and Snijman (1998).
Traub’s scheme included Alliaceae, Hemerocallidaceae,
and Ixioliriaceae as subfamilies, following Hutchinson
(1934, 1959) in part. Within his subfamily Amarylloideae, he erected two informal taxa, ‘‘infrafamilies’’
Amarylloidinae and Pancratioidinae, both of which were
polyphyletic (Meerow, 1995). Dahlgren, Clifford, and
Yeo (1985) dispensed with any subfamilial classification
above the level of tribe, recognizing eight, and treated as
Amaryllidaceae only those genera in Traub’s Amarylloideae. Stenomesseae and Eustephieae were combined.
Meerow (1995) resurrected Eustephieae from Stenomesseae and suggested that two new tribes may need to be
recognized, Calostemmateae and Hymenocallideae. Müller-Doblies and Müller-Doblies (1996) recognized ten
tribes (among them Calostemmateae) and 19 subtribes,
many of them monogeneric; Meerow and Snijman (1998)
recognize 14 tribes, with two subtribes only in one of
them (Table 1).
Fay and Chase (1996) recircumscribed Amaryllidaceae
to include Agapanthus, previously included in Alliaceae,
as subfamily Agapanthoideae. This recircumscription was
based on phylogenetic analysis of rbcL sequence data,
with only four genera of Amaryllidaceae s.s. included in
the analysis. All the epigynous genera were treated as
Amaryllidoideae. Bootstrap support for this treatment
was weak (63%). The sampling within Amaryllidaceae
OF
BOTANY
[Vol. 86
s.s. in Fay and Chase (1996) did not allow sufficient resolution of the generic relationships within the family, and
we present here phylogenetic analyses of three plastid
DNA sequence data sets for a much wider range of taxa.
The phylogenetic application of sequences of rbcL is
well documented (e.g., Chase et al., 1993; Olmstead and
Palmer, 1994) and has been used to clarify relationships
between and within a number of asparagoid families, including Themidaceae (Fay and Chase, 1996), Asphodelaceae (de Bruijn et al., unpublished data), Alliaceae (Fay
et al., unpublished data), and Orchidaceae (Cameron et
al., 1999). Within Amaryllidaceae, however, levels of resolution obtained within some major clades, particularly
those from the Neotropics, were not sufficient to elucidate
tribal relationships fully (Fay et al., 1995). For this reason, we chose to combine our rbcL matrix with two for
the trnL intron/trnL-F spacer region of noncoding plastid
DNA, for which Taberlet et al. (1991) had developed
‘‘universal’’ primers for amplification. Sequences of this
region have been used in phylogenetic studies of Crassulaceae (Kim, t’Hart, and Mes, 1996; Mes, Van Brederode and t’Hart, 1996; Mes, Wijers, and t’Hart, 1997),
Gentianaceae (Gielly and Taberlet, 1996; Gielly et al.,
1996), Paeoniaceae (Sang, Crawford and Stuessy, 1997),
Proteaceae (Maguire et al., 1997), Ranunculaceae (Kita,
Ueda, and Kadota, 1995), among others, either alone or
in combination with other loci. This region of the plastid
genome evolves more than three times faster, on average,
than rbcL (Gielly and Taberlet, 1994) and can therefore
potentially add increased resolution to a phylogeny generated by rbcL sequences.
Combining independent character matrices, whether
both molecular or molecular and morphological, very often increases the resolution of the ingroup and the bootstrap support of the internal nodes of the phylogenetic
trees (Chase et al., 1995b; Olmstead and Sweere, 1994;
Rudall et al., 1998; Soltis et al., 1998). In this paper we
present the first family-wide phylogenetic analysis of
Amaryllidaceae using three plastid DNA sequences,
alone and in combination, and comment on the evolutionary and bigeographic implications of the results.
MATERIALS AND METHODS
Plant materials—The sources of plant material and vouchers/accessions used in this analysis are listed in Table 2, along with GenBank or
EMBL accession numbers for the sequences.
DNA extraction, gene amplification, and sequencing—Sequences
for rbcL were generated at both RBG Kew and the University of Florida
(Table 2); all trnL-F sequences were obtained at Kew.
RBG Kew—DNA was extracted from 1.0 g fresh, 0.2–0.25 g silica
gel-dried leaves, or ;0.1 g material from herbarium sheets using the
2X CTAB method of Doyle and Doyle (1987). All samples were then
purified on cesium chloride/ethidium bromide gradients (1.55 g/mL density). Gene amplification of the rbcL gene was carried out using forward
primers that match the first 20 or 26 base pairs (bp) of the coding region
and reverse primers that correspond to 20-bp sequences that begin at
position 1352 or 1367 in the coding region (Table 3; Chase and al.,
1995a). The trnL-trnF region was amplified using the c and f primers
of Taberlet et al. (1991). Amplified products were purified using Magic
mini columns (Promega, Madison, Wisconsin) or QIAquick (Qiagen,
Valencia, California) columns, following manufacturers protocols. Stan-
September 1999]
MEEROW
ET AL.—AMARYLLIDACEAE SYSTEMATICS
1327
TABLE 1. Four most recent intrafamilial classifications of Amaryllidaceae s.s. The lines indicate subsequent segregation or inclusion of genera.
a As Dahlgren, Clifford, and Yeo (1985) did not consistently list the component genera in their tribal concepts, their exact generic composition
is inferred. Most of their delimitations are presumed to have followed Traub (1963).
dard dideoxy methods or modified dideoxy cycle sequencing with dye
terminators run on an ABI 373A or 377 automated sequencer (according
to the manufacturer’s protocols; Applied Biosystems, Inc., Foster City,
California) were used to sequence the amplification products directly.
For rbcL, both strands were sequenced for 70–90% of the exon. We
have ;1320 bp of rbcL sequence data for most taxa. The trnL-F region
is length variable; we sequenced both strands for 70–90% of the region,
obtaining between 750 and 900 bp of sequence data for most taxa.
University of Florida—Genomic DNA was obtained from young
fresh leaf tissue using an extraction protocol developed for plant tissues
rich in soluble polysacchrides (Li, Cai, and Guy, 1994). Polymerase
chain reaction (PCR) protocols were those of Li and Guy (1996). The
PCR reaction product was fractionated by electrophoresis on a 0.8%
low melting point agarose gel, then the DNA was purified from gel
slices with phenol and chloroform and dissolved in 10 mL of 10 mmol/L
Tris (pH 8.0) buffer. A 10-mL ligation reaction was prepared containing
1328
TABLE 2. Taxa, voucher specimens, and GenBank accession numbers used in the plastid DNA sequence phylogeny analyses of Amaryllidaceae.
GenBank accession number
Taxon
Alliaceae
Allium siculum var. bulgaricum
A. subhirsutum L.
Gilliesia graminea Lindl.
Ipheion uniflorum (Graham) Raf.
Leucocoryne pauciflora R. Phil.
Milula spicata Prain
Nothoscordum bivalve Britton
Pabellonia incrassata (Phil.) Quezada and Martic.
Solaria atropurpurea (Phil.) Rav.
Stemmatium narcissoides Phil.
Voucher
rbcL
trnL gene
trnL-F spacer
GBAN-Z69206
GBAN-Z69203
GBAN-AF117023
GBAN-AF116999
GBAN-AF117051
GBAN-AF117030
Agapanthaceae
Agapanthus africanus Hoffm.
Agapanthus campanulatus F. M. Leighton
M. W. Chase 627 (K)
M. W. Chase 1008 (K)
GBAN-Z69221
GBAN-Z69220
GBAN-AF117028
GBAN-AF117029
GBAN-AF117060
GBAN-AF117059
Amaryllidaceae
Amaryllis belladonna L.
Apodolirion lanceolatum Benth. and Hook.
Boophone disticha (L. f.) Herb.
Brunsvigia comptonii W. F. Barker
Caliphruria korsakoffii (Traub) Meerow
Calostemma lutea Sims
Chlidanthus fragrans Herb.
Clivia nobilis Lindl.
Crinum yemense Deflers
Cryptostephanus vansonii Verdoom
Cyrtanthus elatus (Jacq.) Traub
Eucharis castelnaeana (Baill.) Macbr.
Eucrosia eucrosioides (Pax) Traub
Eustephia darwinii Vargas
Galanthus plicatus M. Bieb.
Gethyllis ciliaris (Thunb.) Thunb.
Griffinia hyacinthina Ker Gawler
Habranthus martinezii Ravenna
Haemanthus humilis Jacq.
Hannonia hesperidium Braun-Blanq. and Maire
Hessea zeyheri Baker
Hieronymiella marginata (Pax) A. T. Hunz.
Hippeastrum papilio (Rav.) Van Scheepen
Hymenocallis marginata (Pax.) A. T. Hunz.
Ismene longipetala (Lindl.) Meerow
Ismene narcissiflora Jacq.
Ismene vargasii (Velarde) Gereau and Meerow
Lapiedra martinezii Lag.
Leptochiton quitoensis (Herb.) Sealy
Leucojum autumnale L.
Lycoris squamigera Maxim.
Narcissus elegans (Haw.) Spach
Nerine bowdenii Will. Wats.
M. W. Chase 612 (K)
Kirstenbosch, NBG 714/88
M. W. Chase 2246 (K)
M. W. Chase 2240 (K)
M. W. Chase 962 (K)
M. W. Chase 1505 (K)
Meerow 2312 (FLAS)
M. W. Chase 3080 (K)
M. W. Chase 1595 (K)
Meerow 2310 (FLAS)
M. W. Chase 1572 (K)
Schunke 14156 (FLAS)
Meerow 1117 (FLAS)
M. W. Chase 559 (K)
M. W. Chase 741 (K)
Duncan 1123 (NBG)
Meerow 2106 (FLAS)
M. W. Chase 1023 (K)
M. W. Chase 2025 (K)
M. W. Chase 2023 (K)
M. W. Chase 2238 (K)
M. W. Chase 1901 (K)
Meerow 2307 (FLAS)
M. W. Chase 1901 (K)
M. W. Chase 3583 (K)
Meerow 2306 (FLAS)
Meerow 2308 (FLAS)
M. W. Chase 1528 (K)
Meerow 1116 (FLAS)
M. W. Chase 607 (K)
M. W. Chase 2014 (K)
M. W. Chase 617 (K)
M. W. Chase 616 (K)
GBAN-Z69219
GBAN-AF116944
GBAN-AF116945
GBAN-AF116946
GBAN-AF116947
GBAN-AF116948
GBAN-AF116949
GBAN-AF116950
GBAN-AF116951
GBAN-AF116952
GBAN-AF116953
GBAN-AF116954
GBAN-AF116955
GBAN-AF116956
GBAN-Z69218
GBAN-AF116957
GBAN-AF116958
GBAN-AF116959
GBAN-AF116960
GBAN-AF116961
GBAN-AF116962
GBAN-AF116963
GBAN-AF116964
GBAN-AF116965
GBAN-AF116966
GBAN-AF116967
GBAN-AF116968
GBAN-AF116969
GBAN-AF116970
GBAN-Z77256
GBAN-AF116971
GBAN-AF116972
GBAN-AF116973
GBAN-AF10479
GBAN-AF104789
GBAN-AF104801
GBAN-AF104813
GBAN-AF104810
GBAN-AF104790
GBAN-AF104770
GBAN-AF104776
GBAN-AF104784
GBAN-AF104804
GBAN-AF104818
GBAN-AF104798
GBAN-AF104788
GBAN-AF104794
GBAN-AF104799
GBAN-AF104816
GBAN-AF104771
GBAN-AF104772
GBAN-AF104781
GBAN-AF104812
GBAN-AF104813
GBAN-AF104807
GBAN-AF104775
GBAN-AF104796
N/A
GBAN-AF104787
GBAN-AF104802
GBAN-AF104806
GBAN-AF104779
GBAN-AF104773
GBAN-AF104780
GBAN-AF104791
GBAN-AF104769
GBAN-AF104744
GBAN-AF104767
GBAN-AF104726
GBAN-AF104722
GBAN-AF104731
GBAN-AF104740
GBAN-AF104723
GBAN-AF104763
GBAN-AF104756
GBAN-AF104743
GBAN-AF104753
GBAN-AF104766
GBAN-AF104742
GBAN-AF104727
GBAN-AF104730
GBAN-AF104745
GBAN-AF104736
GBAN-AF104738
GBAN-AF104721
GBAN-AF104734
GBAN-AF104741
GBAN-AF104757
N/A
GBAN-AF104719
GBAN-AF104768
GBAN-AF104725
GBAN-AF104732
GBAN-AF104750
GBAN-AF104755
GBAN-AF104758
GBAN-AF104733
GBAN-AF104746
GBAN-AF104751
Tristagma bivalve (Lindl.) Traub
Tulbaghia violacea Harv.
[Vol. 86
GBAN-AF117057
GBAN-AF117058
GBAN-AF117045
GBAN-AF117049
GBAN-AF117053
GBAN-AF117056
GBAN-AF117052
GBAN-AF117054
GBAN-AF117050
GBAN-AF117048
BOTANY
GBAN-AF117001
GBAN-AF117000
GBAN-AF117018
GBAN-AF117021
GBAN-AF117025
GBAN-AF117002
GBAN-AF117024
GBAN-AF117026
GBAN-AF117022
GBAN-AF117020
OF
GBAN-Z69200a
GBAN-Z69205
GBAN-Z69208
GBAN-AF116992
GBAN-AF116998
GBAN-AF116991
GBAN-Z69202
GBAN-Z69209
GBAN-Z69207
N/A
AMERICAN JOURNAL
M. W. Chase 835 (K)
M. W. Chase 439 (K)
M. W. Chase 450 (K)
M. W. Chase 627 (K)
UC Irvine Arboretum 8182
Grey-Wilson & Phillips 752 (K)
M. W. Chase 247 (NCU)
UCI Arboretum 8247
M. W. Chase 693 (K)
Beckett, Cheese & Watson 4688
(NY)
M. W. Chase 692 (K)
M. W. Chase 248 (NCU)
GenBank accession number
Taxon
rbcL
trnL gene
trnL-F spacer
GBAN-AF116974
GBAN-AF116975
GBAN-AF116976
GBAN-AF116977
GBAN-AF116978
GBAN-AF116979
GBAN-AF116980
GBAN-AF116981
GBAN-AF116982
GBAN-Z69217
GBAN-AF116983
GBAN-AF116984
GBAN-AF116985
GBAN-AF116986
GBAN-AF116987
GBAN-AF116988
GBAN-AF116989
GBAN-AF116990
GBAN-AF104814
GBAN-AF104778
GBAN-AF104777
GBAN-AF104809
GBAN-AF104785
GBAN-AF104805
GBAN-AF104782
GBAN-AF104783
GBAN-AF104808
GBAN-AF104800
GBAN-AF104811
GBAN-AF104793
GBAN-AF104819
GBAN-AF104792
GBAN-AF104797
GBAN-AF104786
GBAN-AF104774
GBAN-AF104815
GBAN-AF104759
GBAN-AF104718
GBAN-AF104764
GBAN-AF104729
GBAN-AF104762
GBAN-AF104735
GBAN-AF104720
GBAN-AF104754
GBAN-AF104728
GBAN-AF104739
GBAN-AF104724
GBAN-AF104747
GBAN-AF104765
GBAN-AF104748
GBAN-AF104749
GBAN-AF104760
GBAN-AF104761
GBAN-AF104737
Anthericaceae
Anthericum liliago
Echeandia sp.
Leucocrinum montanum Nutt. ex A. Gray
M. W. Chase 515 (K)
M. W. Chase 602 (K)
M. W. Chase 795 (K)
GBAN-Z69225
GBAN-Z69225
GBAN-Z77252
GBAN-AF117005
GBAN-AF117014
GBAN-AF117003
GBAN-AF117033
GBAN-AF117039
GBAN-AF117031
Behniaceae
Behnia reticulata Didr.
Goldblatt 9273 (MO)
GBAN-Z69226
GBAN-AF117007
GBAN-AF117035
Convallariaceae
Aspidistra elatior Blume
Liriope platyphylla F. T. Wang & T. Tang
Peliosanthes sp.
Polygonatum hookeri Baker
M.
M.
M.
M.
GBAN-Z77269
GBAN-Z77271
GBAN-Z77272
GBAN-Z73695
GBAN-AF117016
GBAN-AF117009
GBAN-AF117006
GBAN-AF117010
GBAN-AF117044
GBAN-AF117038
GBAN-AF117034
GBAN-AF117036
Hemerocallidaceae
Geitonoplesium cymosum
Adelaide B. G. 880709
GBAN-AF116997
GBAN-AF117027
GBAN-AF117055
Hyacinthaceae
Albuca shawii Baker
Hyacinthus orientalis L.
Ornithogalum longebracteatum Jacq.
M. W. Chase 1012 (K)
M. W. Chase 1503 (K)
M. W. Chase 1507 (K)
GBAN-Z69223
GBAN-AF116995
GBAN-Z69224
GBAN-AF117012
GBAN-AF117013
GBAN-AF117008
GBAN-AF117042
GBAN-AF117043
GBAN-AF117037
Laxmanniaceae
Eustrephus latifolius R. Br.
Adelaide B. G. 880587
GBAN-AF116996
GBAN-AF117004
GBAN-AF117032
Themidaceae
Bessera elegans Schult.
Brodiaea jolonensis Eastw.
Milla magnifica E. Moore
Muilla maritima S. Wats.
M. W. Chase 626 (K)
M. W. Chase 1831 (K)
Meerow 2309 (FLAS)
M. W. Chase 779 (K)
GBAN-Z69215
GBAN-AF116993
GBAN-AF116994
GBAN-Z69213
GBAN-AF117015
GBAN-AF117017
GBAN-AF117011
GBAN-AF117019
GBAN-AF117040
GBAN-AF117046
GBAN-AF117041
GBAN-AF117047
a
W.
W.
W.
W.
Chase
Chase
Chase
Chase
833 (K)
1102 (K)
497 (K)
847 (K)
1329
The prefix GBAN has been added for linking the online version of American Journal of Botany to GenBank and is not part of the actual GenBank accession number.
ET AL.—AMARYLLIDACEAE SYSTEMATICS
Meerow 2304 (FLAS)
Meerow 1142 (FLAS)
Meerow 2303 (FLAS)
M. W. Chase 1834 (K)
Meerow 1118 (FLAS)
M. W. Chase 1573 (K)
M. W. Chase 1908 (K)
M. W. Chase 549 (K)
M. W. Chase 577 (K)
M. W. Chase 1591 (K)
Meerow 1159 (FLAS)
M. W. Chase 615 (K)
Snijman 281 (NBG)
Meerow 2301 (FLAS)
M. W. Chase 3640 (K)
M. W. Chase 1066 (K)
Meerow 2302 (FLAS)
M. W. Chase 1836 (K)
MEEROW
Pamianthe peruviana [Stapf]
Pancratium canariensis L.
Paramongaia weberbaueri Valarde
Phaedranassa dubia (HBK) Macbr.
Proiphys cunninghamii (Ait. ex Lindl.) Mabb.
Rauhia decora Ravenna
Rhodophiala moelleri (R. Phil.) Traub
Scadoxus cinnabaerinus (Decne.) I. Friis and I. Nordal
Sprekelia formosissima (L.) Herb.
Stenomesson pearcei Bak.
Stenomesson variegatum (R. and P.) Bak.
Sternbergia lutea (L.) Spreng.
Strumaria truncata Jacq.
Traubia modesta (R. A. Phil.) Ravenna
Ungernia flava Boiss. ex Haussk. ex Boiss.
Vagaria parviflora Herb.
Worsleya rayneri (Hook.) Traub and Moldenke
Zephyranthes filifolia Herb. ex Baker
Voucher
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TABLE 2. Continued.
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TABLE 3. PCR and sequencing primers for rbcL and trnL-F used in
this study.
Sequence
rbcL
Royal Botanic Gardens, Kew
59
ATGTCACCACAAACAGAAAC39
59
GCGTTGGAGAGAGATCGTTTTCT39
59
TCGCATGTACCYGCAGTTGC39
59
CTTTCCAAAATTTCACAAGCAGCA39
University of Florida
59
ATGTCACCACAAACAGAAACTAAAGCAAGT39
59
AATTTGATCTCCTTCCATATTTCGCA39
59
AAACTTTCCAAGGCCCGC39
59
GCGACTTCGGTCTTTTTC39
GGTAAACTGGAAGGGGAA39
59
GCGGGCCTTGGAAAGTTT39
59
trnL-F (Taberlet et al., 1991)
59
CGAAATCGGTAGACGCTACG39
59
ATTTGAACTGGTGACACGAG39
59
GGGGATAGAGGGACTTGAAC39
59
GGTTCAAGTCCCTCTATCCC39
Primer name
1F
636F
724R
(moncots)
1368R
Zurawski’s
Z-1
Zurawski’s
Z-1375R
ALM1
(5Z-427)
ALM2
ALM3
ALM4
trnL-c
trnL-f
trnL-d
trnL-e
25 ng of pGEM-Tt vector (Promega, Madison, Wisconsin), 50–100 ng
PCR products, 1 mL of 103 ligase buffer [300 mmol/L Tris-HCl, pH
7.8, 100 mmol/L MgCl2, 100 mmol/L dithiothreitol, 5 mmol/L ATP, and
1.5 U T4 DNA ligase (Promega, Madison, Wisconsin)]. The ligations
were incubated at 168C overnight. One hundred microlitres of competent E. coli XL-1 Blue cells were transformed with 2.5 mL of each
ligation mixture, and spread on a Luria-Bertani (LB) agar plate (100 3
15 mm) containing 50 mg/mL of ampicillin and 12.5 mL tetracycline.
The plate was spread with 50 mL of 2% X-gal and 50 mL of 100 mmol/
L isopropyl-beta-D-thiogalactopyranoside before using. The plate was
incubated at 378C overnight. Individual colonies were counted, and
white colonies were selected to grow overnight at 378C in LB media
containing 50 mg/mL ampicillin. Plasmid DNA was digested with Pst
I and Sst II, and the restricted DNAs were fractionated on a 0.8%
agarose gel to verify the presence of the cloned insert. Plasmid DNA
containing the cloned, amplified insert was purified, and DNA sequencing was accomplished using the Taq DyeDeoxyy Terminator Cycle
Sequencing Kit (Applied Biosystems Inc., Foster City, California) on
an automated sequencer (Applied Biosystems Inc., Foster City, California) by the DNA sequencing Core of the Interdisplinary Center for
Biotechnology Research at the University of Florida. Sequencing was
accomplished using vector T7 and Sp6 primers along with specific primers for the rbcL gene received from G. Zurawski (Table 3). The complete sequence of both strands was determined using sets of synthetic
primers (Table 3).
Sequence alignment—Sequences of rbcL were easily aligned manually because no length variation was detected. For trnL-F, two methods
were employed. Sequences of several taxa representing the range of
probable variation in the matrix were aligned using the Clustal option
in Sequence Navigator (Applied Biosystems, Inc.), followed by manual
optimisation and alignment of subsequent sequences. Alternatively, the
program Sequencher (Gene Codes, Inc., Ann Arbor, Michigan) was used
to align sequences of closely related taxa with subsequent builds of
these smaller alignments performed manually. Copies of the aligned
matrices are available from the senior author.
Analysis—Aligned matrices were analyzed using the parsimony al-
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gorithm of the software package PAUP* for Macintosh (v4.0 d59-64,
Swofford, 1998) with a successive weighting (SW; Farris, 1969) strategy. SW was employed to globally reduce the effect of highly homoplasious base positions on the resulting topologies (Lledó et al., 1998;
Wenzel, 1997). Whole category weights (codon or tranversion) exhibit
broad and overlapping ranges of consistency (Olmstead, 1997), whereas
SW independently assesses each base position of the multiple alignment
based on their consistency in the initial analysis. The initial tree search
was conducted under the Fitch (equal weights; Fitch, 1971) criterion
with 1000 random sequence additions and SPR (subtree pruning-regrafting) branch swapping but permitting only ten trees to be held at
each step to reduce the time spent searching trees at suboptimal levels.
All trees collected in the 1000 replicates were swapped on to either
completion or an upper limit of 5000 trees. The characters were then
reweighted by the rescaled consistency index, and a further 50 replications of random sequence additions were conducted with the weighted
matrix saving 15 trees per replication. These trees were then swapped
on to completion or an upper limit of 5000 trees. The resulting trees
were then used to reweight the matix a second time by the rescaled
consistency index, and another 50 replications of random sequence addition conducted, saving 15 trees per replication, with subsequent swapping on those trees. This cycle was repeated until two successive rounds
found trees of the same length. All analyses were run with the MULPARS option and ACCTRAN optimization. Branches with zero length
were collapsed if the maximum value 5 0 (‘‘amb 1’’). Internal support
was determined by bootstrapping (5000 replicates) with the final reweighted character matrix and with the jackknife program (5000 replicates) of Farris et al. (1996) without SW weights applied. The cut-off
bootstrap percentage is 50; minimum jackknife support percentage is
63 (Farris et al., 1996). The rbcL matrix consisted of 81 taxa, 51 Amaryllidaceae s.s. representing 48 genera, and 30 additional taxa representing 28 genera of Agapanthaceae, Alliaceae, Anthericaceae, Behniaceae, Convallariaceae, Hyacinthaceae, Laxmanniaceae, and Themidaceae, with Geitonoplesium sp. (Hemerocallidaceae) used as outgroup.
The trnL-F matrix includes these same with the addition of Stemmatium
narcissoides (Alliaceae).
The trnL-F region consists of an intron, a short exon, and an intergene spacer (Taberlet et al., 1991). We combined the components of
trnL-F because they are nearly all noncoding, but each of the two larger
regions was analyzed separately to determine whether they were congruent. Because they were congruent (results not shown), we lumped
them together as the ‘‘noncoding matrix’’ to compare directly with rbcL
before we combined all of them.
RESULTS
The rbcL matrix alone—Of 1340 included base positions in the analyses, 226 were parsimony informative.
More than 5000 equally most parsimonious Fitch trees
were found (tree length 5 974) with a consistency index
(CI) of 0.62 and a retention index (RI) of 0.71. SW produced at least 5000 equally parsimonious trees with a
length of 450819 (Fitch length 5 975), a CI 5 0.88 (Fitch
5 0.62), and RI 5 0.89 (Fitch 5 0.70). The large number
of equally parsimonious trees is largely the result of the
short branch lengths that occur within most of the internal
clades (Figs. 1–2) and the imposed constraints against
collapsing zero-length branches. However, the strict consensus of the weighted trees is more resolved than that
of the Fitch trees. The additional step of the SW trees is
essentially the ‘‘cost’’ of optimizing consistent characters
over highly homoplastic base positions (Lledó et al.,
1998). The Amaryllidaceae are not resolved as monophyletic in the strict consensus of all 5000 SW trees; in
these Agapanthus and Amaryllidaceae tribe Amaryllideae
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ET AL.—AMARYLLIDACEAE SYSTEMATICS
1331
Fig. 1. One of 5000 equally parsimonious trees generated by cladistic analysis of the successively weighted rbcL sequence matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface) percentages are
below branches supported by one or both. An asterisk below a branch signifies that both bootstrap and jacknife 5 100%. A white bar across a
branch signifies lack of resolution in the strict consensus tree of the 5000 trees. ‘‘Agapanthus afr.’’ 5 A. africanus, ‘‘Agapanthus cam.’’ 5 A.
campanulatus, ‘‘Allium sub.’’ 5 A. subhirsutum, ‘‘Allium sic.’’ 5 A. siculum var. bulgaricum. The tree is continued in Fig. 2.
form a polytomy with the rest of Amaryllidaceae sensu
stricto (s.s.). Moreover, these clades have no bootstrap or
jackknife support.
The rbcL matrix (Fig. 1) resolves Hyacinthaceae/
Themidaceae as sister to Anthericaceae/Behniaceae with
moderate bootstrap and jackknife support and positions
this clade as sister to Agapanthus/Amaryllidaceae but
with no jackknife or bootstrap support. The Alliaceae are
resolved as an unsupported paraphyletic grade.
In many of the trees (Fig. 1), the African tribe Amaryllideae is sister to the rest of Amaryllidaceae s.s. This
monophyletic group has high bootstrap and jackknife
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Fig. 2. One of 5000 equally parsimonious trees generated by cladistic analysis of the successively weighted rbcL sequence matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface) percentages are
below branches supported by one or both. A white bar across a branch signifies lack of resolution in the strict consensus tree of the 5000 trees.
The tree is continued in Fig. 1.
support (Fig. 1). The rest of the family forms a polytomy
(Fig. 2) that includes a baccate-fruited clade (Haemantheae, including Gethyllideae), the Cyrtantheae (confined
to Africa), Calostemmateae (Australasia), and a monophyletic Eurasian/American group. Of these latter, only
the Eurasian/American clade has any bootstrap (62) and
jackknife support (67). Calostemmateae have a bootstrap
percentage of 63 but no jackknife support. Within the
Haemantheae, Apodolirion and Gethyllis (Gethyllideae)
are resolved as sister taxa in the Fitch topologies, but not
in the SW trees (Fig. 2).
Within the Eurasian/American clade, the American
genera are monophyletic in all trees (Fig. 2) but lack
bootstrap and jackknife support. The Eurasian genera
form a polytomous grade within this clade. These sequences resolve the tribe Hippeastreae (excluding Griffinia and Worsleya 5 Griffineae Ravenna emend.) and
Hymenocallideae. Tribe Hippeastreae are the only clade
in Amaryllidaceae s.s. other than tribe Amaryllideae that
is supported by bootstrap and jackknife percentage greater than 90% (Fig. 2). Worsleya appears as sister to Chlidanthus (Eustephieae), and Griffinia is unresolved (Fig.
2). A subclade representing subtribe Zephyranthinae is
supported by low bootstrap percentage, but the remaining
relationships within this clade are unresolved. The rest of
the American tribes (Eucharideae, Eustephieae, and Stenomesseae) are not resolved by rbcL, but one unexpected
clade with weak bootstrap support (55) encompasses all
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ET AL.—AMARYLLIDACEAE SYSTEMATICS
1333
Fig. 3. One of 5000 equally parsimonious trees generated by cladistic analysis of successively weighted trnL-F sequence matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface) percentages are
below branches supported by one or both. An asterisk below a branch signifies that both bootstrap and jacknife 5 100%. ‘‘Agapanthus afr.’’ 5 A.
africanus, ‘‘Agapanthus cam.’’ 5 A. campanulatus, ‘‘Allium sub.’’ 5 A. subhirsutum, ‘‘Allium sic.’’ 5 A. siculum var. bulgaricum. The tree is
continued in Fig. 4.
the included petiolate-leafed Andean taxa with 2n 5 46
chromosomes.
The trnL-F matrix alone—Of the 1389 base positions
(including gaps) included in the analysis, 378 were parsimony informative. More than 5000 equally most parsimonious trees were found of length 5 1540 with CI 5
0.66 and RI 5 0.73. SW found more than 5000 equally
parsimonious trees of length 5 747723 (Fitch 5 1541)
with CI 5 0.89 (Fitch 5 0.66) and RI 5 0.91 (Fitch 0.73),
the strict consensus of which is more resolved than the
initial Fitch consensus. The trnL-F matrix (Fig. 3) resolves a monophyletic Amaryllidaceae s.s. (bootstrap and
jackknife support . 80%) as sister to Alliaceae with low
bootstrap (56%) and somewhat higher jackknife support
(71%). Agapanthus is sister to the Amaryllidaceae/Alliaceae clade with supporting bootstrap and jackknife percentages of 81 and 87%, respectively.
In all trnL-F topologies the well-supported Amaryllideae is sister to the rest of Amaryllidaceae, with high
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Fig. 4. One of 5000 equally parsimonious trees generated by cladistic analysis of successively weighted trnL-F sequence matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface) percentages are
below branches supported by one or both. A white bar across a branch signifies lack of resolution in the strict consensus tree of the 5000 trees.
The tree is continued in Fig. 3.
bootstrap and jackknife support. As with rbcL, the remaining African tribes (Haemantheae, Gethyllideae, Cyrtantheae) and Australasian Calostemmateae (itself, a
well-supported clade) form an unresolved polytomy with
the American/Eurasian taxa in the strict consensus (Fig
4). Unlike the rbcL topology, Gethyllideae resolves as a
well-supported monophyletic subclade of Haemantheae
that is sister to Haemanthus (Fig. 4).
Hannonia and Lycorideae (Lycoris and Ungernia) are
outside of an otherwise monophyletic Eurasian clade in
which Galantheae (Galanthus and Leucojum) are resolved with high bootstrap (93%) and jackknife (92%)
percentages (Fig. 4). The monophyletic Lycorideae form
a weak clade with Griffinia and Hannonia as sister genera.
Compared to the rbcL topology, the American genera
are less resolved by trnL-F; Griffinia and Worsleya appear outside the clade comprising all other American taxa
September 1999]
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ET AL.—AMARYLLIDACEAE SYSTEMATICS
1335
Fig. 5. One of 5000 equally parsimonious trees generated by cladistic analysis of successively weighted combined rbcL and trnL-F sequence
matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface)
percentages are below branches supported by one or both. An asterisk below a branch signifies that both bootstrap and jacknife 5 100%. ‘‘Agapanthus afr.’’ 5 A. africanus, ‘‘Agapanthus cam.’’ 5 A. campanulatus, ‘‘Allium sub.’’ 5 A. subhirsutum, ‘‘Allium sic.’’ 5 A. siculum var. bulgaricum.
The tree is continued in Fig. 6.
(Fig. 4). The petiolate Andean clade, which appears in the
rbcL consensus, loses two members, Eucharis and Rauhia. Hymenocallideae are not resolved, and Leptochiton
and Pamianthe are resolved as sister genera with moderate bootstrap and strong jackknife support. Hippeastreae (less Griffineae) appear with low bootstrap support
(61) but with different internal resolution than with rbcL.
Again, short branch lengths are characteristic of most of
the internal nodes of the American clade (Fig. 4).
The combined matrix—More than 5000 equally most
parsimonious trees were found of length 5 2546 with CI
5 0.64 and RI 5 0.71. SW found more than 5000 equally
parsimonious trees of length 5 1194297 (Fitch 5 2546)
with CI 5 0.89 (Fitch 5 0.64) and RI 5 0.89 (Fitch 5
0.71). The strict consensus of the weighted trees is more
resolved than the initial Fitch consensus. Agapanthus is
sister to Amaryllidaceae in the combined topologies (Fig.
5), albeit with low bootstrap support (60%). A monophy-
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Fig. 6. One of 5000 equally parsimonious trees generated by cladistic analysis of successively weighted combined rbcL and trnL-F sequence
matrix for Amaryllidaceae and other Asparagalean genera. Numbers above branches are branch lengths. Bootstrap (plain) and jackknife (boldface)
percentages are below branches supported by one or both. The tree is continued in Fig. 5.
letic Alliaceae is sister to the former clade, with a bootstrap of 79% and jackknife of 77%. Both Amaryllideae
and Haemantheae are well-supported tribal clades (Figs.
5–6), with higher bootstrap and jackknife percentages
than in either of the separate analyses, and the former
resolves as sister to the rest of Amaryllidaceae s.s.
Within Amaryllideae, most of the included genera resolve in a grade with Amaryllis and then Crinum as the
successive sister taxa to the rest (Fig. 5). Within Haemantheae, a well-supported, monophyletic Gethyllideae
is again sister to Haemanthus (Fig. 6). As in both the
individual analyses, Calostemmateae and Cyrtantheae remain as part of the polytomy inclusive of Haemantheae
and the large Eurasian/American clade.
In the combined analysis, the Neotropical (American)
and Eurasian genera are sister groups with strong boot-
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ET AL.—AMARYLLIDACEAE SYSTEMATICS
strap and jackknife support (95, 98%), but of the two,
only the American clade has weak bootstrap support (Fig.
6). Galanthus/Leucojum, Hannonia/Vagaria, and Pancratium/Sternbergia are supported sister genera, but the
remaining relationships, consistent in all trees, have no
support.
Within the American clade (Fig. 6), a distinct Andean
subclade has weak bootstrap support (68). Eustephieae
have no consensus, bootstrap, or jackknife support. A
well-supported Hippeastreae are in a polytomy with Griffinia, Worsleya, and the Andean clade. In Hippeastreae,
a distinct Zephyranthinae and Rhodophiala/Traubia are
well supported. Within the Andean clade, the resolution
of Hymenocallideae, observed in the rbcL trees (Fig. 2),
is lost, with Ismene, however, remaining monophyletic.
Leptochiton and Pamianthe are weakly supported sister
taxa. Eucharis and Rauhia fail to join the rest of a weakly
supported petiolate-leafed subclade that is sister group to
Hymenocallis (marked by a single synapomorphy).
DISCUSSION
Suprafamilial relationships of Amaryllidaceae—Fay
and Chase (1996) presented a smaller rbcL analysis and
argued for the inclusion of Agapanthus as a monotypic
subfamily within Amaryllidaceae. The sister-group status
of Agapanthus to Amaryllidaceae s.s. is only weakly supported by our combined matrix (bootstrap 5 60%). Based
on these data, it would be possible to argue for recognizing Amaryllidaceae in a modified Hutchinsonian
(1934) sense, i.e., with three subfamilies, Allioideae,
Agapanthoideae, and Amarylloideae.
Backlund and Bremer (1998) discussed the issue of
monogeneric families and how best to treat them. They
generated a set of guiding principles for classification:
(1) primary principle of monophyly and (2) a set of secondary principles: (a) maximizing stability, (b) maximizing phylogenetic information (5 minimizing redundancy), (c) maximizing support for monophyly, and (d) maximizing ease of identification. Principle 1 is considered
most important by Backlund and Bremer (1998), as it is
by most modern systematists. However, the secondary
principles will vary in importance among different taxa.
Monophyly is maximized by either treating Agapanthus as a monogeneric family or accepting Amaryllidaceae in the Hutchinsonian sense. However, the support
for a broad concept of Amaryllidaceae (including Alliaceae and Agapanthaceae) is only moderate (bootstrap 5
79%, jackknife 5 77%). The only morphological character that unites all three families is the pseudo-umbellate
inflorescence (homoplasious with Themidaceae), whereas
the Alliaceae are readily marked by their solid styles and
sulfonated compounds, and the Amaryllidaceae have inferior ovaries and unique alkaloid chemistry. Maximizing
stability in this case seems rather moot, given that Agapanthus has been maintained for years as part of Alliaceae, a classification that violates the primary principle
of monophyly, and Amaryllidaceae and Alliaceae have
been united on and off again over the last two centuries.
However, maximizing phylogenetic information and ease
of identification are best served by treating Agapanthus
as the sole genus of a separate family (Agapanthaceae
1337
Voight), while maintaining the independent status of Alliaceae.
The combined analysis supports most of the other relationships hypothesized by Fay and Chase (1996). The
sister-group status of Themidaceae and Hyacinthaceae is
confirmed with good support, and there is bootstrap support in the combined analysis for this clade as sister
group to Amaryllidaceae/Alliaceae/Agapanthaceae, although Antheridaceae/Behniaceae resolves outside of this
clade. It should be noted that, with this level of sampling
of the broader Asparagales, these relationships are largely
a matter of outgroup selection.
Relationships within Amaryllidaceae—Within Amaryllidaceae s.s., several groups are well supported within
all of the analyses, some of which correspond to traditionally accepted tribes of the family. The most unexpected resolution concerns the sister status of the Eurasian/American clades. This is only supported in the
combined analysis, and resolution of this group in relation to the remaining African and Australasian clades is
still elusive because of short branch lengths in this portion of the trees (Figs. 2, 4, 6).
In a survey of internal morphology of American and
African Amaryllidaceae, Arroyo and Cutler (1984) noted
several characters that separated American genera from
African. All American species surveyed have scapes with
collenchyma, a one-layered rhizodermis, and obvolute
bracts. All Amaryllideae (entirely African with the exception of pantropical Crinum) have schlerenchyma in
the scape, a multilayered rhizodermis, and equitant
bracts. Haemanthus and Cyrtanthus exhibit scape and
root anatomy of the American species but the equitant
bracts of Amaryllideae (Arroyo and Cutler, 1984). Calostemmateae (Calostemma and Proiphys), which were
not discussed by Arroyo and Cutler (1984), have equitant
bracts. Many of the Eurasian genera have fused spathe
bracts, which obscures the pattern of their coherence, but
both Lycoris and Pancratium species with free bracts
show the equitant condition. Obvolute bracts may thus
be a synapomorphy of the American clade.
Two American subclades are found in the consensus
of the combined analysis (Fig. 5), with both Griffinia and
Worsleya forming a polytomy with them. The more
weakly supported Andean subclade (tribes Eucharideae,
Stenomesseae and Eustephieae) is characterized by 2n 5
46 chromosomes, which has been interpreted as a tetraploid derivation from an ancestral 2n 5 22 (Meerow,
1985, 1987a, c, 1989). The strongly supported Hippeastreae is characterized for the most part by x 5 6 or 11,
with diploid chromosome numbers of 22, 24 or less. The
short branch lengths and numerous polytomies in the Andean group (Fig. 6) may indicate that they are a relatively
young clade with an evolutionary history tied closely to
the geologically recent Andean uplift (Meerow, 1987c).
Four recognized tribes of Amaryllidaceae are consistently resolved by the plastid DNA sequences, and all
receive strong bootstrap and jackknife support in at least
the combined analysis. These are the Amaryllideae, Haemantheae, Calostemmateae, and Hippeastreae.
Amaryllideae—This tribe, with much of its generic diversity confined to South Africa is sister to the rest of
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the Amaryllidaceae and has high bootstrap and jackknife
support. Compared to other tribes in Amaryllidaceae,
Amaryllideae are marked by a large number of synapomorphies (Snijman and Linder, 1996): extensible fibers
in the leaf tissue, bisulculate pollen with spinulose exines,
scapes with a sclerenchymatous sheath, unitegmic or
ategmic ovules, and nondormant, water-rich, nonphytomelanous seeds with chlorophyllous embryos. A few of
the genera extend outside of South Africa proper, but
only Crinum, with seeds well adapted for oceanic dispersal (Koshimizu, 1930), ranges through Asia, Australia,
and America. Snijman and Linder’s (1996) phylogenetic
analysis of the tribe based on morphological, seed anatomical, and cytological data resulted in recognition of
two monophyletic subtribes: Crininae (Boophone, Crinum, Ammocharis, and Cybistetes) and Amaryllidinae
(Amaryllis, Nerine, Brunsvigia, Crossyne, Hessea, Strumaria, and Carpolyza). Müller-Doblies and Müller-Doblies (1996) recognized four subtribes with little discussion and no phylogenetic analysis: Crininae, Boophoninae, Amaryllidinae, and Strumariinae, the latter two containing several segregate genera from Hessea and
Strumaria (Table 1). Our sampling of this tribe is incomplete, and therefore we feel it is premature to attach a
great deal of confidence to the generic sister relationships
seen here. Four genera of Snijman and Linder’s (1996)
Amaryllidinae do form a weakly supported clade (Fig. 5)
with Amaryllis as sister to the rest of the tribe in the rbcL
and combined analyses. Identical positioning of Amaryllis occurred in Snijman’s (1992) cladistic analyses if tribe
Hippeastreae was used as the outgroup, with Haemantheae as outgroup (Snijman and Linder, 1996), and also
both outgroups used (Snijman, 1992). Amaryllis resolves
as sister to a clade containing the other genera they ultimately placed, with Amaryllis, in subtribe Amaryllidinae. Müller-Doblies and Müller-Doblies’ (1996) concept
of Amaryllidinae [Amaryllis, Nerine, and Namaquanula
(5 Hessea)] would make their subtribe Strumariinae paraphyletic and Amaryllidinae polyphyletic.
Haemantheae—This baccate-fruited tribe is another
morphologically well-marked group with strong molecular support. The limits of the tribe, however, have been
controversial. Müller-Doblies and Müller-Doblies (1996)
insisted on retaining Cyrtanthus in the tribe, albeit as a
monotypic subtribe, Cyrtanthinae. The basis for uniting
Cyrtanthus with the Haemantheae has always been weak,
chiefly the shared chromosome number with Haemanthus
(2n 5 16; Ising, 1970; Vosa and Snijman, 1984) and its
strictly African range. This diploid number also occurs
in some Hippeastreae (Flory, 1977; Grau and Bayer,
1991). Uniting Cyrtanthus with Haemantheae has no molecular support in our analyses, and we believe that Cyrtanthus, the only solely African genus with the flattened,
winged, phytomelanous seed so common in the American
clade, should be recognized as a monotypic tribe (Traub,
1963; Dahlgren, Clifford, and Yeo, 1985; Meerow and
Snijman, 1998). Recognition of Gethyllideae as a distinct
tribe (Müller-Doblies and Müller-Doblies, 1996; Meerow
and Snijman, 1998), however, is not supported by the
molecular data. Although the large, elongate, baccate
fruits and small hard seeds of Apodolirion and Gethyllis
are a departure from the berries and large succulent seeds
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of the rest of Haemantheae, the two genera, though resolved as sister taxa (Figs. 4, 6), are firmly embedded
within Haemantheae. Recognizing them as a distinct taxon would render the rest of Haemantheae paraphyletic.
Haemantheae are the only tribe of Amaryllidaceae that
contain rhizomatous genera (Cryptostephanus and Scadoxus in part), a condition that occurs in the sister family
Agapanthaceae. This has generally been conceived as a
plesiomorphy within the family (Nordal and Duncan,
1981; Meerow, 1995, 1997; Müller-Doblies and MüllerDoblies, 1996). In the rbcL and combined consensus
trees, the three ‘‘bulbless’’ genera form a grade at the
base of the Haemantheae, which would support this hypothesis, although Cryptostephanus, the only member of
the tribe with the ancestral state of a phytomelanous testa,
is not the first branch in the grade. Haemanthus and Scadoxus, which have been treated as one genus in the past
(e.g., Hutchinson, 1934, 1959; Traub, 1963), are sister
genera only in the rbcL topologies (Fig. 2). The position
of Scadoxus, the only genus of the tribe polymorphic for
the rhizomatous state, as the final terminal taxon in the
‘‘bulbless’’ grade seems reasonable. In any event, all
three matrices render recognition of a subtribe Cliviinae
for Clivia and Cryptostephanus by Müller-Doblies and
Müller-Doblies (1996) as paraphyletic. Any further insight on the internal relationships within Haemantheae
requires additional sampling.
Calostemmateae—Calostemmateae, treated as part of a
polyphyletic Eucharideae by Hutchinson (1934, 1959),
Traub (1963), and Dahlgren, Clifford, and Yeo (1985),
were first suggested as a distinct lineage by Meerow
(1989) and formally recognized by Müller-Doblies and
Müller-Doblies (1996). The tribe consists of two Australasian genera (Proiphys, forest understory herbs of Malaysia, Indonesia, the Philippines and tropical Australia,
and Calostemma, endemic to Australia). A few species
of Crinum, with the broadest distribution of any genus in
the family, are the only other members of Amaryllidaceae
present in Australia. The indehiscent capsules of both
genera are similar in appearance to the unripe berry-fruits
of Scadoxus and Haemanthus (Haemantheae), but early
in the development of the seed, the embryo germinates
precociously, and a bulbil forms within the capsule and
functions as the mature propagule (Rendle, 1901). The
two genera exhibit the equitant bract condition of the
African and Eurasian genera.
Hippeastreae—All but two of the genera treated by
Meerow and Snijman (1998) as part of Hippeastreae are
resolved as a well-supported monophyletic clade in all
the analyses (Figs. 2, 4, 6). The two genera that lie outside of this clade are Worsleya and Griffinia, both Brazilian endemics, exhibiting the rare character of bluerange pigmentation in the flowers. The variable positioning of these two in the various analyses is interesting in
itself. In the trnL-F topologies (Fig. 4), Worsleya is part
of the basal polytomy within the Eurasian/American
clade, whereas Griffinia weakly resolves as sister to the
Mediterranean Hannonia (the latter on a long terminal
branch). In the rbcL consensus (Fig. 2), Worsleya resolves as sister to Chlidanthus (Eustephieae), whereas
Griffinia remains unresolved along with the rest of Eus-
September 1999]
MEEROW
ET AL.—AMARYLLIDACEAE SYSTEMATICS
tephieae. In the combined analysis (Fig. 6), both are positioned within the American clade, but unresolved with
either Hippeastreae s.s. or the weakly supported tetraploid
Andean clade. The failure of Worsleya or Griffinia to
resolve as part of Hippeastreae in any of the analyses
casts doubt on Müller-Doblies and Müller-Doblies’
(1996) submergence of Worsleya in Hippeastrum and
weakens Meerow and Snijman’s (1998) retention of both
genera in tribe Hippeastreae.
Another unexpected indication of relationship occurs
within the tetraploid Andean clade, where a distinct petiolate-leafed subclade is resolved in the rbcL topologies
(Fig. 2). This resolution is not retained by the trnL-F and
combined analyses in which Eucharis and Rauhia are
pulled from this group. Nonetheless, a core of petiolate
genera remain monophyletic, with weak support in the
combined analysis. Despite the fact that petiolate leaves
have evolved independently several times elsewhere in
the Amaryllidaceae (Amaryllideae, Calostemmataceae,
Haemantheae, Hymenocallideae, and Hippeastreae), the
molecular data begin to indicate that it may be a synapomorphy for this group. Hymenocallideae as a distinct
tribe receive weak support (55%, one synapomorphy) in
the rbcL matrix only, and the rest of Stenomesseae is
poorly resolved by all matrices.
Within the Eurasian clade of the combined analysis
(Fig. 6), Lycorideae appears as sister to the rest, although
without support. This tribe represents the more or less temperate Asian component of the family, with Lycoris ranging from Korea, through China, Myanamar, and Japan, and
Ungernia restricted to the mountains of central Asia. Müller-Doblies and Müller-Doblies (1978) described similarities in the bulb anatomy of Ungernia and Sternbergia, a
possible synapomorphy between Lycorideae and the rest
of this clade. One genus of the Eurasian clade, Pancratium, is represented throughout Africa, tropical Asia, as well
as Mediterranean Europe and the Middle East, a distribution that could signify a more ancestral position within
the Eurasian clade. The current data, however, do not support this resolution for Pancratium, with only a single species from the Canary Islands represented in the analyses.
The presence of Pancratium in Africa may thus be secondary, though it is the only genus outside of the African
tribes Amaryllideae and Haemantheae with external trichomes (Björnstad, 1973).
Sister relationships of Hannonia and Vagaria receive
good bootstrap and jackknife support, as does the traditional alliance of Galanthus and Leucojum. Crespo et al.
(1995), using ITS sequences, refuted Müller-Doblies and
Müller-Doblies’ placement of Lapiedra in Pancratieae,
and our data support a closer relationship with Galantheae or Narcisseae for this genus. However, concepts of
Galantheae, Narcisseae, and Pancratieae presented in
Müller-Doblies and Müller-Doblies (1996) or Meerow
and Snijman (1998) are not resolved in any of the three
analyses, and we believe that caution should be used before categorical statements are made about tribal lineages
within this group.
The low internal branch lengths throughout the Amaryllidaceae, except in some of the deepest branches, are
a striking contrast to the other asparagalean families included in the analysis (Figs. 1, 3, 5). The significance of
this is not clear. It could mean that a great deal of the
1339
modern diversity in the family is of relatively recent occurrence (as is likely, for example, within the Andean
clade), or else base substitution rates in the chloroplast
genome are lower within the family than for other Asparagales.
Character state evolution in the Amaryllidaceae—By
optimizing morphological or other ‘‘traditional’’ characters onto a gene tree, one is able to gain insight about
putative transformation series or state polarities that have
characterized the evolution of the group under study. This
can be useful for constructing a character state matrix for
an ingroup in which rampant homoplasy in such characters confounds the endeavor. Certain characters that
have been used to justify older intrafamilial classifications of Amaryllidaceae do show stability within some
of the clades resolved by the combined analysis, while
others appear extremely homoplasious (Fig. 7).
The most notable correlation between evolutionary
depth as resolved by plastid DNA sequences and morphological synapomorphies is found in the Amaryllideae
(Fig. 7). All of the characters listed are synapomorphous
for the tribe, which terminates the longest internal branch
within Amaryllidaceae on our gene trees (Figs. 1, 3, 5).
Presence or absence of bulbs—The bulbless condition
occurs in the sister group to Amaryllidaceae, the monogeneric Agapanthaceae. It is also the character state for
the only South African subfamily of Alliaceae, Tulbaghoideae (Fay and Chase, 1996). In Amaryllidaceae, the
absence of bulbs characterizes only three genera, Clivia,
Cryptostephanus, and Scadoxus, but the latter also includes species that form a true bulb. If this is a symplesiomorphy as most have interpreted it (Nordal and Duncan,
1984; Müller-Doblies and Müller-Doblies, 1996; Meerow
and Snijman, 1998), the bulbous state has evolved at least
three times in the family, in Amaryllideae, Haemantheae,
and within the ancestral stock for the rest of the family.
Petiolate leaves—Petiolate or, more accurately, pseudopetiolate leaves are widepread throughout the Asparagales, and this character exhibits a great deal of homoplasy within Amaryllidaceae (Meerow and Snijman,
1998). At the extreme, one-to-few petiolate species occur
in otherwise lorate-leafed genera (e.g., Crinum, Hymenocallis). The state may occur throughout a genus, but
renders a tribe polymorphic (Calostemmateae, Haemantheae, Griffineae). In the tetraploid Andean clade, a subclade is defined by the synapomorphy of a petiolate leaf
in the rbcL trees, but Eucharis and Rauhia pull away with
trnL-F and in the combined analyses.
Mesophyll palisade—It has been suggested that the
presence or absence of a distinct palisade layer in the leaf
mesophyll may have systematic significance (Arroyo and
Cutler, 1984; Artyushenko, 1989). Petiolate-leafed taxa
never have palisade chlorenchyma (Meerow and Snijman, 1998). It is characteristic of Amaryllideae (Crinum
is polymorphic), but absent in Haemantheae (the state in
unknown for Gethyllis and Apodolirion). In Calostemmateae, it is present in Calostemma but absent in the
petiolate Proiphys (Meerow, unpublished data). Palisade
almost universally occurs in the Eurasian clade. It is ab-
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AMERICAN JOURNAL
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[Vol. 86
Fig. 7. Amaryllidaceae/Agapanthaceae clade from one of 5000 equally parsimonious trees generated by cladistic analysis of the successively
weighted combined rbcL and trnL-F sequence matrix for Amaryllidaceae and other Asparagalean genera with selected morphological and karyological states optimized on the tree. A ‘‘P’’ to the right of a character bar or box refers to the state being polymorphic in the adjacent taxon; if
superimposed on the character bar itself, polymorphy is widespread among all adjacent taxa. ‘‘Agapanthus afr.’’ 5 A. africanus, ‘‘Agapanthus
cam.’’ 5 A. campanulatus, Ismene E 5 I. subg. Elisena, Ismene I 5 I. subg. Ismene, Ismene P 5 I. subg. Pseudostenomesson, Stenomesson p. 5
S. pearcei, Stenomesson var. 5 S. variegatum.
sent in Leucojum (Artyushenko, 1989), and Galanthus is
polymorphic (Davis and Barnett, 1997). Within the
American clade, it is wholly characteristic of the Eustephieae (Arroyo and Cutler, 1984; Meerow and Snijman,
1998) but occurs only sporadically within the Hippeas-
treae (Arroyo and Cutler, 1994). The state of Worsleya is
not known. Outside of Eustephieae, a distinct palisade is
absent from the Andean tetraploid clade (Meerow, 1987a,
1989). The inference based on the distribution of this
character state on our topology (Fig. 7) is that a distinct
September 1999]
MEEROW
ET AL.—AMARYLLIDACEAE SYSTEMATICS
palisade is plesiomorphic within the family, though the
state within Agapanthaceae, sister to Amaryllidaceae, has
not to our knowledge been reported.
Pubescence—The presence of trichomes on the external parts of Amaryllidaceae is common only in some
Amaryllideae and Haemantheae and one African species
of Pancratium (Arroyo and Cutler, 1984; Meerow and
Snijman, 1998). It is completely unknown in the American clade, Cyrtantheae, and Calostemmateae. It may
have evolved independently in the three clades within
which it occurs.
Scape characters—Solid scapes are the predominant
condition in Amaryllidaceae as occurs in Agapanthaceae
as well. Hollow scapes are almost universally characteristic of Hippeastreae, and thus appears to be a synapomorphy for that tribe. The only other genera within which
hollow scapes occur are Leucojum and Cyrtanthus both
of which are polymorphic for the character (Traub, 1963;
Reid and Dyer, 1984). As discussed previously, obvolute
spathe bracts seem to be apomorphic for the American
clade, and the presence of schlerenchyma in the scape is
an autapomorphy for Amaryllideae.
Floral symmetry—Zygomorphic and actinomorphic
flowers occur in the Amaryllidaceae, and several genera
(Crinum, Cyrtanthus, Phycella) are polymorphic. Snijman and Linder (1996) consider actinomorphy the apomorphic condition in Amaryllideae. The flowers of Agapanthaceae are zygomorphic. Within Haemantheae, only
Clivia is zygomorphic. In the American clade, zygomorphy is the rule in the ‘‘hippeastroid’’ subclade. Pyrolirion
and Zephyranthes (including Haylockia) are the only genera characterized exclusively by actinomorphic flowers,
while Phycella (not included in the sequence analyses) is
polymorphic. In the Andean subclade, only Eucrosia,
Plagiolirion, and Ismene subgenus Elisena are exclusively zygomorphic; Rauhia is polymorphic. The Eurasian
clade is on the whole actinomorphic; only Lycoris is
characterized by zygomorphic flowers. The mosaic occurrence of actinomorphy throughout the family (Fig. 7)
and the occurrence of polymorphic genera suggest that
transformations between the two states of floral symmetry may be easily modified by pollinator-mediated selection, and perhaps controlled by one or few genes.
Paraperigone—The ‘‘paraperigone’’ is an anomalous
secondary outgrowth of the perianthal meristem with
ramifying vasculature (Arber, 1939; Singh, 1972), not to
be confused with a similar-looking structure formed by
staminal connation (see below). It is most well developed
(and typified) by the corona of Narcissus. Such a welldeveloped paraperigone occurs in only one other genus,
the Chilean endemic Placea (Hippeastreae). However, a
homologous series of fimbrae, scales, or a continuous callose ring occurs in Cryptostephanus (Haemantheae), one
or two species of Cyrtanthus, and variably thoughout Lycorideae and Hippeastreae. It has thus probably evolved
at least three times (Haemantheae, Cyrtantheae, and the
Eurasian/American clade), but from a meristematic potential that is deep rooted in the family. Polymorphism
1341
for this character within genera may suggest that it is
easily lost.
Staminal connation—The fusion of the staminal filaments was the single most important character with
which Traub (1957, 1963) justified recognizing his ‘‘infrafamily’’ Pancratioidinae, a subfamilial taxon that in
fact was glaringly polyphyletic. Though staminal connation is a widespread character state within the Andean
tetraploid clade (Fig. 7), it is paralleled elsewhere in the
family, particularly in Amaryllideae subtribe Amaryllidinae (Snijman and Linder, 1996), the Calostemmateae,
in Gethyllis, and some species of Cyrtanthus (Reid and
Dyer, 1984). In the Eurasian clade it occurs in Pancratium, the flower morphology in general of which bears
striking resemblance to several Andean genera (Hymenocallideae pro parte, Paramongaia, and Pamianthe).
Meerow and Dehgan (1985) attemepted to link these socalled ‘‘pancratioid’’ genera by pollen morphology, but a
more parsimonious explanation may be convergence for
pollinator specificity (Morton, 1965; Bauml, 1979; Grant,
1983). However, Pancratium and these Andean genera
are monophyletic in a larger sense (as part of the Eurasian/American clade), and the exact position of Pancratium within the Eurasian subclade is still not strongly
resolved (Fig. 6).
Fruit and seed characters—Fruit and seed morphology
have been an important focus of experimentation within
the family. Baccate fruits have apparently evolved only
once, despite the difference in gross morphology between
the long, aromatic fruit of Gethyllis and Apodolirion and
the berries of the rest of Haemantheae. Phytomelan [the
ancestral state for all Asparagales (Huber, 1969)] has
been lost from the testa as many as five times in the
Amaryllidaceae: in Amaryllideae, Griffineae, Hymenocallideae, Haemantheae, and Calostemmateae [in Calostemmateae a true seed never forms, but an integumentary
rudiment is present (Rendle, 1901)]. In both Haemantheae and Hymenocallideae, phytomelan is found around
the seeds of one genus each (Cryptostephanus and Leptochiton, respectively). The loss of one integument [or
both, as been controversially reported for some Crinum
(Prillieux, 1858; von Schlimbach, 1924; Tomita, 1931;
Markötter, 1936, but see Snijman and Linder, 1996)] is
synapomorphic for Amaryllideae.
A flattened, winged seed, which occurs in Agapanthaceae, is very common in the American clade, but otherwise occurs only in Ungernia (Lycorideae) and Cyrtantheae. The most similar type of seed to this is the Dshaped seed of Worsleya and some Pancratium. A dry,
hard, wedge-shaped or irregularly round seed is characteristic of most of the Eurasian clade (except Lycorideae),
frequently with an elaiosome at the chalazal end. Among
all genera of the family, Pancratium is the most polymorphic for seed type (Werker and Fahn, 1975).
Characterization of certain seeds of Amaryllidaceae as
fleshy (regardless of whether phytomelan is present) has
led, in the past, to false homologies (see discussion in
Meerow, 1989). Truly fleshy seeds occur in Amaryllideae
(in which case the bulk of the seed volume is endosperm;
Rendle, 1901), Hymenocallideae (the fleshy portion is integumentary; Whitehead and Brown, 1940), and some
1342
AMERICAN JOURNAL
water-rich Haemantheae. But an ‘‘intermediate’’ state occurs in a number of genera in which the seed is round,
turgid, but not really fleshy (the seed will burst under
pressure rather than give way), and contains copious, oily
endosperm. This type of morphology is found in Cryptostephanus (Haemantheae), Lycoris (Lycorideae), Eucharideae sensu Meerow (1989), Griffinia, and a single
species of Hippeastrum.
Chromosome number—A chromosome number of 2n
5 22 is considered plesiomorphic in Amaryllidaceae due
to the broad occurrence in many of the tribes of the family (Goldblatt, 1976; Flory, 1977; Meerow, 1984, 1987b).
Andean-centered genera in the tribes Eucharideae, Eustephieae, Hymenocallideae, and Stenomesseae are characterized by a somatic chromosome number of 2n 5 46
or presumptive derivations thereof (Di Fulvio, 1973, Flory, 1977; Williams, 1981; Meerow, 1987a, b). Resolution
of these genera as a clade in our plastid DNA trees supports the interpretation of a monophyletic polyploid origin for these tribes from an ancestor with 2n 5 22 via
chromosome fragmentation or duplication and subsequent doubling or vice versa (Satô, 1938; Lakshmi, 1978;
Meerow, 1987b).
Biogeographic implications—Raven and Axelrod
(1974) postulated a western Gondwanaland origin for
Amaryllidaceae sensu Huber (1969), and this is supported by the plastid DNA phylogeny. The deepest branches
of the topology originate in Africa, including the sister
group of the family Agapanthus. Africa has also been the
site of considerable innovation in the family’s history as
well, as typified by the Afrocentric tribes Amaryllideae,
Haemantheae, and Cyrtantheae. Most of the diversity
within those three tribes is, however, centered in South
Africa, and thus may reflect radiation engendered by the
more recent paleoclimatic and geological history of Africa encompassing Neogene and later times (Axelrod,
1972; Raven and Axelrod, 1974). The increased aridity
of the African climate and the uplift of the continental
mass beginning near the end of the Oligocene, further
abetted by Quaternary climatic fluctuations, were catastrophic to many elements of the African flora, but it may
have been a selective pressure for diversity among groups
of geophytes capable of adapting to increasing drought.
The geophyte richness of South Africa is well documented (Goldblatt, 1978), and the Cape region has been
suggested as a possible refuge for certain African plant
and animal groups as the tropical flora of the continent
was impoverished (Raven and Axelrod, 1974). However,
the three basal genera of the baccate-fruited Haemantheae
according to our combined analysis (Fig. 6), Clivia,
Cryptostephanus, and Scadoxus, are all forest understory
taxa, do not form bulbs, and are at least in part (Scadoxus,
Cryptostephanus) elements of tropical vegetation farther
north. Cryptostephanus does not occur in South Africa at
all, and this is the only genus of Haemantheae in which
the plesiomorphic state of a phytomelanous testa occurs.
The Calostemmateae, the only exclusively Australasian
element of the family, may have been isolated from the
African lineages as Australia separated from western
Gondwanaland (Raven and Axelrod, 1974). Direct migration between Africa and Australia may have persisted
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[Vol. 86
up through the close of the early Cretaceous, although
India and Madagascar may have provided a less direct
corridor up until the late Cretaceous (Raven and Axelrod,
1974). That the Calosternmateae remains within the unresolved grade of otherwise African tribes would suggest
relative antiquity for the lineage. Crinum is the only
amaryllid that is known to occur on Madagascar, despite
the island’s probable role as a refuge for taxa decimated
by the Neogene African extinctions, whereas indigenous
Indian amaryllids are restricted to Crinum and two to
three species of Pancratium. The adaptations of Crinum
for long-distance dispersal have been demonstrated (Koshimizu, 1930), and Pancratium may have been able to
directly enter India from either Africa or Eurasia during
the late Cretaceous or early Eocene (Raven and Axelrod,
1974).
The sister relationship of the Eurasian/Mediterranean
clade to the American genera raises the interesting question of when and where the Amaryllidaceae, in the main,
entered the New World. It should be noted that this probably occurred at least twice, as the arrival of Crinum in
the Americas via oceanic dispersal was undoubtedly an
unrelated event (Arroyo and Cutler, 1984). Although migration between Eurasia and North America has been
possible throughout most of angiosperm history (Raven
and Axelrod, 1974), the hypothesized pathways have
been for plants of temperate forest biota and not considered to be important for plants of subhumid or semiarid
vegetation (Raven, 1971, 1973). However, eastern North
America and western Europe may have shared a warm,
seasonally dry climate from the late Cretaceous to the
early Eocene (Axelrod, 1973, 1975), which might have
allowed east/west movement of species, with island
chains of the Mid-Atlantic ridge providing stepping
stones. Such a Madrean-Tethyan hypothesis would have
the initial entry of the Amaryllidaceae into the New
World through North America. Although there are members of the family in Mexico and the southern United
States, they are, with the exception of the ubiquitous Crinum, components of terminal subclades (Zephyranthes,
Habranthus, Hymenocallis) in an overall American phylogeny based on nuclear DNA ITS sequences (Meerow,
Guy, and Li, 1998), all of which are linked to more basal
taxa endemic to South America. The validity of the Madrean-Tethan hypothesis has more recently been questioned by various studies using isozyme, plastid DNA
restriction fragment length polymorphisms (RFLPs), or
cladistic analyses of taxa considered emblematic of the
disjunction: Buxus (Köhler and Brückner, 1989), Datisca
(Liston, Rieseberg, and Hanson, 1992), Lavatera (Ray,
1994), Quercus (Manos, 1992; Nixon, 1993), Pinus (Little and Crutchfield, 1969; Miller, 1993), and Styrax
(Fritsch, 1996). In these cases, the hypothesized Madrean-Tethyan linkage is not resolved as monophyletic, the
taxon itself is not monophyletic, or the estimated time of
divergence does not fit the Madrean-Tethyan hyothesis.
Given the extant distribution of Amaryllidaceae in
North America and the generic richness south of the
equator, a northern latitude entry into the New World for
the family would necessitate massive extinction in North
America sometime after migration to South America took
place. Glaciation would be the likely factor involved. Little migration of plants from North America to South
September 1999]
MEEROW
ET AL.—AMARYLLIDACEAE SYSTEMATICS
America probably took place before the Eocene (Raven
and Axelrod, 1974). All indications are that the movement of extant Amaryllidaceae has been northward from
South America (e.g., Meerow, 1987b, 1989). This does
not necessarily preclude an earlier, initial arrival in North
America, migration to South America, and a more recent,
but secondary, return of some elements of the family to
North America long after glaciation extirpated the founder populations. However, if a North American entry is
hypothesized, this begs the question of why the Eurasian
sister clade has been so successful in adapting to temperate habitats, which constitute the majority of the species in tribes Galantheae, Narcisseae, and Lycorideae,
whereas the American clade is relatively depauperate of
temperate climate adaptation. There is nothing in our data
to prove or disprove an initial New World entry of the
Amaryllidaceae into North America, and the issue is for
the present unresolved.
In conclusion, our combined analysis of plastid DNA
sequences rbcL and trnL-F provide good support for the
monophyly of the Amaryllidaceae and indicate Agapanthaceae as its likely sister family. The Alliaceae are in
turn sister to the Amaryllidaceae/Agapanthus clade. The
origins of the family are African. The phylogenetic relationships with Amaryllidaceae s.s. resolve strongly
along biogeographic lines. The tribe Amaryllideae, primarily South African and well supported by numerous
morphological synapomorphies, is sister to the rest of
Amaryllidaceae. The remaining two African tribes of the
family, Haemantheae and Cyrtantheae, are well supported, but their position relative to the Australasian Calostemmateae and a large clade comprising the Eurasian/
American genera, is not yet clear. The Eurasian elements
of the family and the American genera are monophyletic
sister clades. Internal resolution of the Eurasian clade
only partially supports currently accepted tribal concepts,
and few conclusions can be drawn on the relationships
of the genera based on these data. A monophyletic Lycorideae (Central and East Asian) is weakly supported.
Galanthus and Leucojum (Galantheae pro parte) are supported as sister genera by the Bootstrap. The American
clade shows a higher degree of internal resolution. A
monophyletic Hippeastreae (less Griffinia and Worsleya)
is well supported, and a distinct subtribe, Zephyranthinae,
is resolved as well. A distinct Andean clade marked by
a chromosome number of 2n 5 46 and derivations thereof is resolved with weak support, and a distinct petiolate
Andean subclade composed of elements of the tribes Eucharideae and Stenomesseae is partially resolved with
weak support. The lack of resolution of Griffinia and
Worsleya in the overall American clade, and of Eustephieae in the Andean subclade, may indicate that these
genera represent more isolated elements of the American
lineage.
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