Genet Resour Crop Evol (2009) 56:263–275
DOI 10.1007/s10722-008-9363-5
RESEARCH ARTICLE
The origin of Datura metel (Solanaceae): genetic
and phylogenetic evidence
Mario Luna-Cavazos Æ Robert Bye Æ Meijun Jiao
Received: 2 October 2007 / Accepted: 15 July 2008 / Published online: 19 August 2008
Ó Springer Science+Business Media B.V. 2008
Abstract Using the analysis of nine isozyme sys-
tems and cladistic analysis of 32 morphological
characters of the Mexican species of the section Dutra
of the genus Datura, evidence was sought on the origin
of the cultivated D. metel L. The genetic similarity and
the phylogenetic relationship suggest that D. metel is
related more closely to D. inoxia Mill. than to the other
taxa of the section Dutra based upon the small genetic
distance between them. The cladistic analysis revealed
two main clades: the long-lived, tuberous rooted
perennials (D. inoxia, D. lanosa A.S. Barclay ex
Bye, D. metel, and D. wrightii Regel) and the tap-
rooted annuals (D. discolor Bernh., D. kymatocarpa
A.S. Barclay, D. leichhardtii F.V. Muell. ex Benth.
ssp. pruinosa (Greenm.) Barclay ex Hammer (syn.:
D. pruinosa Greenm.), D. reburra A.S. Barclay).
Datura inoxia is the sister taxon of D. metel next to
which is D. wrightii while D. lanosa is the basal taxon
of this group. The combination of genetic and cladistic
data indicates that D. inoxia is most likely the
progenitor of D. metel.
M. Luna-Cavazos
Programa de Botánica, Colegio de Postgraduados,
Montecillo, Texcoco, Estado de Mexico 56230, Mexico
R. Bye (&)
M. Jiao
Jardı́n Botánico, Instituto de Biologı́a, Universidad
Nacional Autónoma de México, Apdo. Post. 70-226,
04510 Mexico, DF, Mexico
e-mail: rbyeunam@ibiologia.unam.mx
Keywords Datura metel
Domesticated plant

Isozymes
Mexico
Phylogeny
Solanaceae
Introduction
Datura metel is a member of the section Dutra of the
genus Datura. Eight of the ten taxa of this section are
native to Mexico and have been introduced to other
parts of the world: D. discolor, D. inoxia, D. kymato-
carpa, D. lanosa, D. metel, D. pruinosa, D. reburra,
and D. wrightii. The other taxa are endemic: D. leich-
hardtii F.V. Muell. ex Benth. ssp. leichhardtii to
Australia and D. velutinosa Fuentes to Cuba. If one
accepts that D. leichhardtii and D. pruinosa are
conspecific (Haegi 1976; Hammer et al. 1983; Symon
and Haegi 1991), the latter taxon is D. leichhardtii
ssp. pruinosa (Greenm.) Barclay ex Hammer; for
convenience, this trinomial is cited as D. pruinosa in
the text, tables, and figures. Two other sections are
recognized: section Datura with D. ferox L.,
D. quercifolia Kunth, and D. stramonium L. and the
monotypic section Ceratocaulis with D. ceratocaula
Ortega. Brugmansia or the tree datura was once
treated as a section of Datura (Safford 1921) but is
now considered a separate, though closely related,
genus (Lockwood 1973; Persson et al. 1999). Various
species of Datura are known and widely employed for
their medicinal and toxic properties that are based
upon the more than 30 alkaloids (Hegnauer 1973,
1990; Evans 1979; Tétényi 1987; Bye et al. 1991;
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Dethier et al. 1993). Because of their funnel-form,
fragrant nocturnal blooms, such species as D. inoxia,
D. metel, D. stramonium and D. wrightii are culti-
vated as ornamental plants with all but D. metel
known from wild populations (DeWolf 1956).
Datura metel was described by Linneaus (1753) who
cited Asia as the geographic origin of this species.
Dunal’s treatment of Solanaceae in De Candolle’s
Prodromus (1852) indicated: ‘‘In regionibus calidis
Indiae or Americae merid. et Europae australis; in
Mexico, ad Victoria’’. Other authors (Satina and Avery
1959; Fuentes 1980) continued to cite Asia as the
geographic origin of D. metel and recognized its
distribution in Asia, Africa, and the tropical and
subtropical regions of America. Because it is a wide-
spread cultivated plant, its place of origin has been
difficult to determine (Satina and Avery 1959). In
reviewing the history and taxonomy of Datura in
Australia, Symon and Haegi (1991) concluded that the
genus Datura is native to the American continent and
proposed that D. metel is of American origin and was
introduced subsequently by Europeans to various parts
of the world. No wild populations of D. metel are
known. Symon and Haegi’s hypothesis is supported by
floristic research in China and Japan (An-ming 1986)
that states that D. metel is only found under cultivation.
The global distribution of D. metel is broad and
includes Asian, African and American continents
(Satina and Avery 1959; Fuentes 1980; Gentry and
D’Arcy 1986). It is common in warm areas of China
and India (Deb 1979; An-ming 1986; Jain and
Borthakur 1986). In Mexico, it is found in the states
of Veracruz, Tabasco, Campeche, Yucatan, Quintana
Roo, Chiapas and Oaxaca (Luna-Cavazozs et al.
2000). The lack of wild populations, its presence
only under cultivation, and the peculiar characteris-
tics of the plants suggest that D. metel is a product of
domestication. The flower size, the fruit position and
dehiscence pattern, the perennial herbaceous habit,
and the seeds with caruncles indicate that it is a
member of the section Dutra. The double, multicol-
ored corolla of the long-lived flower and the reduced
ornamentation on the normally spiny fruit suggest
that the plant has undergone radical changes which
can be seen as desirable features for an ornamental
plant. Such morphological, physiological, ecological,
and genetic changes differentiate the domesticated
plant from its wild ancestors (Reynolds and Tampio
1983; Zohary and Hopf 1994).
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Genet Resour Crop Evol (2009) 56:263–275
In addition, isozymatic analysis can suggest tax-
onomic and phylogenetic relationships among species
(Soltis and Soltis 1989; Avise 1994, Baverstock and
Moritz 1996). These analyses provide significant
information on the genetic variation found in domes-
ticated species and their wild ancestors in order to
establish genetic relationships between them and as
an approach of identifying progenitors of domesti-
cated plants (Doebley 1989; Zohary and Hopf 1994).
Examples of isozymatic studies in Solanaceae relat-
ing domesticated species and their wild proegenitors
include: Oliver and Martinez-Zapater (1984) in
Solanum L, and Mcleod et al (1983) and Hernán-
dez-Verdugo et al. 2001 in Capsicum L.
Because domestication is based upon the evolu-
tionary process (Darwin 1883; Harlan 1992), the
selection patterns can be deduced using phylogenetic
analysis (Stuessy 1990). Cladistic methods to estab-
lish evolutionary relationships between wild
progenitors and derived domesticated plants have
been employed in the studies of Helianthus (Riese-
berg et al. 1991), Sphenostylis stenocarpa (Potter and
Doyle 1992), Agave fourcroydes (Colunga-Garcı́a
Marin et al. 1999), and Vicia (Leht and Jaaska 2002)
among other taxa (Spooner and Lara-Cabrera 2001).
As part of our study of the diversification and
evolution of Datura, we apply these approaches to
the phenetic analysis of isozymatic data and the
cladistic analysis of morphological characters of the
Mexican species of Datura section Dutra in order to
identify the progenitor of D. metel as well as to reveal
the evolutionary tendencies of this species under the
domestication process. The objectives of this report
are: (1) analyze the genetic relationship of D. metel
with Mexican species of the section Dutra using
phenetic analysis of isozymes, (2) propose a phylo-
genetic relationship of the Mexican taxa of section
Dutra using cladistic analysis of morphological
characters, and (3) suggest the origin of D. metel
and the major evolutionary changes that occurred
under domestication.
Materials and methods
Plant material
Thirty populations of the following species were
sampled (29 throughout Mexico and one from
Genet Resour Crop Evol (2009) 56:263–275 265

Table 1 Sources of plant specimens of Datura section Dutra used in isozyme analysis
Datura discolor
México: Nayarit, R. Bye 13369; Querétaro, Arroyo Seco, Puente de Concá, R. Bye et al. 19032. Veracruz, R. Bye 20016.
Datura inoxia
México: San Luis Potosı́, R. Bye 14384; Querétaro, Arroyo Seco, Puente de Concá, R. Bye et al. 19031; Chihuahua, Casas Grandes,
R. Bye 19371; Puebla, R. Bye 20901.
Datura kymatocarpa
México: Guerrero, R. Bye 19439; Guerrero, R. Bye 15819; Guerrero, Iguala, R. Bye 16607; Guerrero, R. Bye 19305; Guerrero, R. Bye
19371.
Datura lanosa
México: Sonora, Cd. Obregón, Rı́o Yaqui, R. Bye 20758; Sonora, R. Bye 13346; Sinaloa, Badiraguato, orilla del rı́o Badiraguato-
Humayo, R. Bye et al. 20643; Chihuahua, Batopilas, R. Bye 18376.
Datura metel
México: Yucatán, Izamal, Holcá, M. Luna C. 1001 V. Fuentes y F. Basurto; Quintana Roo, Cd. Chetumal, M. Luna C. 1006 V.
Fuentes y F. Basurto; Oaxaca, Salina Cruz, Santa Marı́a Huamelula, M. Luna C. 1029 V. Fuentes y F. Basurto; Oaxaca, San Mateo
del Mar, M. Luna C. 1026 V. Fuentes y F. Basurto; Yucatán, R. Bye s.n.
Datura pruinosa Greenm.
México: Puebla, Tehuacan, camino a Jalpan, R. Bye 13484; Puebla, R. Bye s.n.; Puebla, Tehuacan, R. Bye 20902.
Datura reburra
México: Sinaloa, Mocorito, 1 Km al S de la Estación de microondas El Pinto, R. Bye et al. 20658; Sinaloa, Culiacán, R. Bye 13332;
Sinaloa, R. Bye 19659.
Datura wrightii
USA: California, R. Bye 20882; México: Baja California, R. Bye 21867; Chihuahua, R. Bye 13190.
Herbarium specimens are deposited at MEXU

adjacent USA; Table 1): D. discolor (3), D. inoxia
(4), D. kymatocarpa (5), D. lanosa (4), D. metel (5),
D. pruinosa (3), D. reburra (3), and D. wrightii (3).
In each population, a mature fruit from each of five
individuals was collected and the air-dried seeds were
pooled to form a population sample. After surface
sterilization with 2% sodium hypochlorite, the seeds
were planted in germination trays with sterilized
organic soil and grown in the greenhouse to obtain an
average of 10–20 seedlings for each population.
Isozymes
(b) Histidine-citrate (Stuber and Johnson 1977) for 6-
phosphogluconate dehydrogenase (Pgd, E.C. 1.1.1.44)
and Phosphohexose isomerase (Phi, E.C. 5.3.1.9); (c)
Citrate-histidine (Fildes and Harris 1966) for Diapho-
rase (Dia, E.C. 1.6.4.3), Malate dehydrogenase (Mdh,
E.C. 1.1.1.37) and Malic enzyme (Me, E.C. 1.1.1.40),
and (d) Tris-citrate-borate (Poulik 1957) for Shikimic
acid dehydrogenase (Sad, E.C. 1.1.1.25) and Superox-
ide dismutase (Sod, E.C. 1.15.1.1). The electrophoretic
analysis followed the procedures of Conkle et al.
(1982), Cheliak and Pitel (1984) and Stuber et al.
(1988).

The fresh, fully expanded second leaf of each seedling
was macerated in a solution of sucrose and sodium
ascorbate. The crude extract was placed on strips of
Whatman filter paper (2 9 8 mm), placed in plastic
tubes and stored in a freezer at -70°C until use. The
electrophoresis was carried out in horizontal chambers
on 12% starch gels. Using four different buffer
systems, we analyzed nine isozymes: (a) Lithium-
borate (Ashton and Braden 1961) for Gluta-
mate-oxaloacetate transaminase (Got, E.C. 2.6.1.1)
and Phosphoglucomutase (Pgm, E.C. 2.7.5.1);
Isozyme data analysis
The interpretation of the band patterns of the gels
was based upon Wendel and Weeden (1989). The
different zones of isozymatic activity were consid-
ered distinct loci that were numbered consecutively
from the anode. The bands within each locus were
scored as distinct alleles as described by Cheliak
and Pitel (1984) in which the numbering of alleles is
sequential with the most common allele being
number one.

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The allelic frequencies were estimated using the
procedures of Brown and Weir (1983). A locus was
considered polymorphic if the frequency of the most
common allele was less than 0.95. The genetic
variability between taxa was calculated as follows:
(a) the portion of polymorphic loci where the number
of polymorphic loci of a population and/or species
was divided by the total number of loci; (b) the
average number of alleles per locus in which the total
of alleles was divided by the total number of loci for
the populations and species; (c) the average number
of alleles per polymorphic locus that equals the total
number of alleles divided by total number of
polymorphic loci.
The intraspecific and interspecific genetic similar-
ity in the section Dutra was calculated using the
matrix of allelic frequencies per population and
species, respectively. In the latter case the averages
of allelic frequencies for n populations of each taxon
were used in calculating the Nei’s Coefficient of
Genetic Distance (Nei 1972). The dendrograms
illustrating the intraspecific and interspecific relation-
ships of the taxa were constructed based upon the
values of the Unweighted Pair Group Method of
Arithmetic Averages (UPGMA) using NTSYS-pc
version 2.1 (Rohlf 2000).
Cladistic analysis
In order to understand the evolutionary tendencies of
D. metel, the phylogenetic relationship among the
Mexican taxa in the section Dutra was established
using morphological characters and biochemical
features such as seed oil, starch, and protein for the
following taxa: D. discolor, D. inoxia, D. kymato-
carpa, D. lanosa, D. metel, D. pruinosa, D. reburra,
and D. wrightii. The 32 characters (Table 2) were
scored for the taxa of interest and submitted to
cladistic analysis. The character states were polarized
by an outgroup (Watrous and Wheeler 1981) which in
this case was Brugmansia sanguinea (Ruiz et Pav.)
D. Don. This genus was identified as the sister group
to Datura based upon molecular markers (Olmstead
and Palmer 1992) and has been recognized as
possessing such primitive characters as arborescent
habit, bilocular ovary and autoincompatible breeding
system (Lockwood 1973). Phylogenetic analysis of
pollen characters (Persson et al. 1999) confirms the
distinctiveness between these closely related genera.
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Genet Resour Crop Evol (2009) 56:263–275
Data for the morphological characters were mea-
sured on 5 to 10 samples per population each derived
from a field collection where 10 to 15 individuals per
population were collected; the vegetative portions
were carefully separated, pressed and dried while the
fruits and flowers were preserved in a solution of
alcohol and glycerin. Eight morphological and seed
characters were derived from a parallel study (Car-
mona-Jiménez 2003). Of the 32 characters compiled,
17 have two states, and 15 have multiple states. The
quantitative characters were codified based upon
variation intervals (Stevens 1991). The data matrix of
nine taxa, including the outgroup, with their 32
characters is presented in Table 3. The cladisitic
analysis used the program PAUP 4.0 (Swofford
1998). An exhaustive search was made for the
phylogenetic trees using the option ALLTREES,
and support for the clades was provided by BOOT-
STRAP (Felsenstein 1985).
Results
Isozymes
The nine isozymatic systems analyzed revealed 15
loci and detected 50 alleles (data available upon
request) with an average of 2.167 alleles per locus.
Only the locus Mdh3 was monomorphic for all
populations. Different levels of intraspecific and
interspecific polymorphisms were observed. The
portion of polymorphic loci per population ranged
from 0.645 in D. pruinosa to 0.933 in D. metel; the
average number of alleles per locus varied from 2.0 in
D. reburra in to 2.3 in D. discolor; the average
number of alleles per polymorphic locus ranged from
1.5 in D. pruinosa to 2.1 in D. inoxia (Table 4). The
isozymatic phenotypes observed in Datura sección
Dutra are as follows.
Malate dehydrogenase (Mdh). Three loci were
found in this isozymatic system. Mdh1 was
polymorphic in all species except D. discolor and
D. kymatocarpa. Mdh2 was polymorphic in all
taxa except D. lanosa, while Mdh3 was monomor-
phic in all taxa.
Phosphohexose isomerase (Phi). One locus with
considerable polymorphism is found in all species.
Five alleles appear in different frequencies in the
Genet Resour Crop Evol (2009) 56:263–275 267

Table 2 Characters and

character states used in the
cladistic analysis of Datura
section Dutra with
Brugmansia sanguinea as
outgroup
1. FOVI: life form
2. RAIZ: root
3. Foho: leaf form
4. MAHO: leaf margin
5. PUHO: leaf pubescence
6. LOCA: calyx length
7. VECA: calyx venation
8. LOCO: corolla length
9. LICO: corolla limb
10. COCO: corolla color
11. CGCO: corolla throat color
12. FOCO: corolla form
13. VECO: corolla verticils
14. LOAN: anther length
15. COFI: filament color
16. DEFR: fruit dehiscence
17. CAFR: calyx on fruit
18. PSFR: fruit pseudoseptum
19. ESFR: fruit spines
20. LOSE: seed length
21. COSE: seed color
22. LOTS: testa lobules
23. BATH: border of hilum cup
24. DLSE: lateral depression of seed
25. RETE: testa reticulation
26. ARSE: aril of seed
27. ANTH: angle of hilum
28. ANHM: angle between hilum and
micropyle
29. AASE: abundance of oil in seed
30. ALSE: abundance of oil in seed
31. PRSE: abundance of protein in seed
32. ESME: layers of mesotesta of seed
0 = woody, 1 = herbaceous perennial,
2 = herbaceous annual
0 = napiform, 1 = tap root
0 = ovate, 1 = trullate
0 = entire, 1 = dentate
0 = glabrous, 1 = puberulent, 2 = pilose, 3 = lanate
0 = 8–12 cm, 1 = \5 cm
0 = not pronounced, 1 = pronounced
0 = 25–30 cm, 1 = 15–20 cm, 2 = 10–15 cm,
3 = \10 cm
0 = acumen [ interacumen,
1 = acumen = interacumen
2 = acumen \ interacumen
0 = other than white, 1 = white
0 = other than white, 1 = white
0 = hypocrateriform, 1 = infundibuliform,
2 = campanulate
0 = simple, 1 = double, triple
0 = 30–35 mm, 1 = 10–16 mm, 2 = \10 mm
0 = other than green, 1 = pale green
0 = indehiscent, 1 = irregular, 2 = regular
0 = without calyx, 1 = normal, 2 = reflexed
0 = septum complete, 1 = with septum &
pseudoseptum
0 = absent, 1 = few, 2 = many
0 = [5 mm, 1 = 4–5 mm, 2 = \4 mm
0 = brown, 1 = black
0 = poorly defined, 1 = well defined, 2 = absent
0 = present, 1 = absent
0 = present, 1 = absent
0 = present, 1 = absent
0 = absent, 1 = present
0 = 110–200°, 1 = 210–225°, 2 = 245–250°
0 = 140°, 1 = 75–60°, 2 = 55–40°
0 = scarce, 1 = abundant
0 = scarce, 1 = abundant, 2 = very abundant
0 = scarce, 1 = abundant
0 = 10–20, 1 = 4, 2 = 1–3

species although allele 2 is not present in D. pru-
inosa and D. reburra, allele 3 and 5 are absent in
D. reburra. Only allele 6 is found in D. wrightii.
Glutamate-oxaloacetate transaminase (Got). Two
loci for this isozyme are revealed. Got1 is poly-
morphic for all species except D. reburra, while
Got2 is polymorphic in all the taxa.
Phosphoglucomutase (Pgm). Two loci are found.
Pgm1 with four alleles is polymorphic in all the
species; differences are found in allele Pgm1-3 that
exists in D. discolor, D. kymatocarpa, D. reburra
and D. metel as well as allele Pgm1-4 that is
present only in D. discolor. Locus Pgm2 is poly-
morphic in all species except D. reburra. In the

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Table 3 Character matrix used in cladistic analysis of the species of Datura section Dutra
FOVI RAIZ FOHO MAHO PUHO LOCA VECA LOCO LICO COCO CGCO FOCO VECO LOAN COFI DEFR

B. sanguinea 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
D. discolor 2 1 1 1 0 0 1 2 2 1 0 0 0 2 1 2
D. inoxia 1 0 0 0 1 0 0 1 0 1 1 1 0 1 1 1
D. kymatocarpa 2 1 1 1 0 1 0 3 2 1 0 0 0 2 1 2
D. lanosa 1 0 0 0 3 0 0 1 0 1 1 2 0 1 1 1
D. metel 1 0 0 0 0 0 0 1 0 0 1 1 1 1 0 1
D. pruinosa 2 1 1 1 0 1 0 3 1 1 0 0 0 2 1 2
D. reburra 2 1 0 1 0 1 1 3 1 1 1 0 0 2 1 2
D. wrightii 1 0 0 0 2 0 0 1 0 1 1 2 0 1 1 1
CAFR PSFR ESFR LOSE COSE LOTS BATH DLSE RETE ARSE ANTH ANHM AASE ALSE PRSE ESME

B. sanguinea 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
D. discolor 2 1 1 2 1 2 0 1 1 1 2 1 0 0 1 1
D. inoxia 1 1 1 2 1 0 1 1 1 0 1 1 1 1 1 1 2
D. kymatocarpa 2 1 1 1 0 0 0 0 0 1 2 2 0 2 0 2
D. lanosa 1 1 2 1 0 1 0 1 0 1 2 1 0 1 0 1
D. metel 1 1 0 1 0 1 1 1 0 1 1 2 1 1 0 2
D. pruinosa 1 1 1 2 0 2 1 1 1 1 2 2 1 2 0 2
D. reburra 1 0 1 0 1 1 0 1 0 1 0 1 0 1 0 2
D. wrightii 1 1 2 1 0 1 0 1 0 1 1 2 1 2 0 2
Genet Resour Crop Evol (2009) 56:263–275
Genet Resour Crop Evol (2009) 56:263–275
Table 4 Average genetic variability of 30 populations of eight
Mexican species of Datura section Dutra
Species PL PA
D. discolor 0.733 2.311
D. inoxia 0.883 2.267
D. kymatocarpa 0.747 2.120
D. lanosa 0.850 2.117
D. metel 0.933 2.187
D. pruinosa 0.645 2.089
D. reburra 0.689 2.000
D. wrightii 0.845 2.244
PL: portion of polymorphic loci; PA: average number of alleles
per locus; PAP: average number of alleles per polymorphic
locus
case of Pgm2-3, it is absent in D. pruinosa and
D. reburra while Pgm2-4, Pgm2-5 and Pgm2-6 are
found only in D. discolor.
6-phosphogluconate dehydrogenase (Pgd). This
isozyme system consists of two loci. Locus Pgd1
is polymorphic in all species except D. discolor
and D. pruinosa; allele Pgd1-3 is exclusive to
D. pruinosa, D. inoxia and D. lanosa. Locus Pgd2
has the lowest level of polymorphism of all the
loci. It is polymorphic in all perennial species
group: D. inoxia, D. lanosa, D. metel y D. wrightii
and in only three populations of the annual species
group: one in D. pruinosa and two in D. reburra
with high alleleic frequencies (0.912 in the former
and 0.945 and 0.936 in the latter).
Superoxide dismutase (Sod). One locus with four
alleles is recorded. It is polymorphic in all the
species. Allele 1 is absent in D. discolor while
allele 3 is present in the annual species group
except in D. pruinosa and in D. metel of the
perennial species group.
Diaphorase (Dia). Two polymorphic loci are
registered for this isozyme system. Allele Dia1-1
is common in all the species except D. kymato-
carpa and D. pruinosa. In the case of allele Dia1-2
it is absent in D. discolor. Allele Dia1-3 is
common in the annual taxa and in D. wrightii of
the perennial group. Dia2-2 is found in all taxa
except D. discolor and D. reburra.
Malic enzyme (Me). One locus with two alleles is
present in this isozyme system. It has low levels of
polymorphism with the most common allele vary-
ing from 0.904 to 0.944.
269
Shikimic acid dehydrogenase (Sad). One locus is
found in all the taxa except D. pruinosa. High
PAP allelic frequencies are registered for D. kymato-
carpa and D. reburra of the annual group and for
1.845 D. wrightii of the perennial group. The last species
2.117 is the only one with allele 3.
1.680
1.883
2.080
1.511 Genetic distance
1.534
2.022
The genetic similarity of D. metel compared to other
taxa in the section Dutra is seen in the allelic
frequencies and the genetic distance. Based upon the
allelic frequencies, D. inoxia is most similar to
D. metel and differs (in declining order) only in loci
Phi1, Pgm1 and Sad1. When comparing the values
for genetic distance between D. metel and other
species of the section Dutra (Table 5), the average
genetic distances between it and D. inoxia is 0.037.
Datura lanosa and D. wrightii have slightly higher
indices (0.043; 0.064; respectively). The remaining
species are more distant in the following order (of
increasing average): D. reburra, D. kymatocarpa,
D. discolor, and D. pruinosa.
The dendrogram of genetic distances for the 30
populations of the section Dutra (Fig. 1) indicate
that: (1) the populations of the same species form
compact groups, and (2) two major branches are
recognized in the dendrogram of the genetic dis-
tances. The first group consists of the D. discolor,
D. reburra, D. kymatocarpa, and D. pruinosa, the
last being the most dissimilar from D. metel. The
second group is more differentiated and includes
D. inoxia, D. metel, D. lanosa, and D. wrightii; and
two subgroups are composed of the following species
pairs: D. inoxia–D. metel and D. lanosa–D. wrightii.
Cladistic analysis
Cladistic analysis of the 32 morphological and
biochemical characters resulted in one parsimonious
cladogram, with 71 steps, a consistency index of 0.690
and a retention index of 0.627. Two clades are evident
in the cladogram (Fig. 2): one with bootstrap value of
67% that consists of the long-lived perennial herbs
D. inoxia, D. metel, D. wrightii, and D. lanosa, and
the other with bootstrap support of 60% that includes
the annual herbs D. discolor, D. kymatocarpa,
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270 Genet Resour Crop Evol (2009) 56:263–275

D. pruinosa, and D. reburra. In the first major clade

(with 54% support), D. inoxia shares a common

(0.011–0.015)
D. wrightii

ancestor with D. metel both of which are subsequently

0.013
related to D. wrightii while D. lanosa is located at the
base. In the second major clade (with 77% support),
D. discolor and D. kymatocarpa share a common
ancestor and are related to D. pruinosa while D. re-

(0.005–0.014)

(0.190–0.281)
burra is basal. The perennial species are united by
D. reburra

various synapomorphies defined by the perennial habit

0.010

0.238
(1), a corolla length between 15 and 20 cm (8), the
corolla forms (12): infundibular in the case of
D. inoxia and D. metel and campanulate in the case
of D. wrightii and D. lanosa, the anther length
(0.018–0.037)

(0.160–0.290)

(0.199–0.273)
D. pruinosa

between 10 and 16 mm (14), the irregular fruit

0.026

0.212

0.245
dehiscence (16) and the seed length of 4–5 mm (20).
The clade D. inoxia–D. metel shares the character of
the infundibular corolla (12).
Table 5 Minimum, maximum and average values of the genetic distance between species of Datura section Dutra

(0.008–0.022)

(0.262–0.362)

(0.247–0.332)

(0.052–0.079)

Discussion
D. metel

0.015

0.304

0.282

0.064

Even though recent studies (Symon and Haegi 1991)

suggested that D. metel originated in the American
continent, no probable progenitor was proposed nor
(0.028–0.063)
(0.009–0.026)

(0.201–0.284)

(0.216–0.331)

(0.007–0.037)

was the relationship of D. metel to other members of

D. lanosa

0.016

0.043

0.240

0.268

0.023

the section Dutra clarified.

The first isozyme study (using ten species, six of
which are members of Dutra, and peroxidase enzymes;
Conklin and Smith 1971) concluded that there are five
D. kymatocarpa

(0.015–0.029)

(0.354–0.186)

(0.097–0.246)

(0.066–0.144)

(0.176–0.300)
(0.234–0.371)

groups. Datura metel was derived from a D. discolor

ancestor and, in turn, D. inoxia and D. wrightii
0.022

0.262

0.291

0.156

0.105

0.240

(reported as D. meteloides) were derived from D. metel

ancestors. A study (Fuentes and Lima 1983) using the
same isozyme system and nine herbaceous species and
three woody species (Brugmansia) concluded that it
(0.027–0.051)
(0.016–0.034)

(0.165–0.297)

(0.068–0.018)

(103–0.209) 0.150 (0.195–0.316)

(0.189–0.283)

(0.081–0.042)

was not possible to determine the relationship among

D. inoxia

0.024

0.246

0.038

0.037

0.238

0.242

0.061

the herbaceous species. Because of the limited number

of systems used and the problematic nature of perox-
idase, these studies provide little information (Soltis
and Soltis 1989).
(0.259–0.340)
(0.011–0.033)

(0.205–0.325)

D. kymatocarpa (0.090–0.160)

(0.384–0.244)

(0.068–0.125)

(0.199–0.305)

The AFLP technique applied to eight species of

D. discolor

Datura and six species of Brugmansia using Atropa

0.023

0.266

0.132

0.294

0.293

0.096

0.257

as an outgroup (Mace et al. 1999) identified five

clusters. Members of section Dutra were divided
among Clusters 2 and 3. Datura metel was basal to
Cluster 2 that included D. inoxia and D. wrightii.
D. pruinosa
D. discolor

D. wrightii
D. reburra
D. lanosa
D. inoxia

Cluster 3 included other members of section Dutra

D. metel

(D. inoxia and D. discolor) as well as D. ceratocaula

(of section Ceratocaulis). Even though D. metel is

123
Genet Resour Crop Evol (2009) 56:263–275 271

Fig. 1 Dendrogram D.discolor

illustrating the intraspecific D.discolor
genetic relationship among D.discolor
D.reburra
the 30 populations of D.reburra
Datura section Dutra based D.reburra
upon the Coefficient of D.kymatocarpa
D.kymatocarpa
Genetic Distance of Nei D.kymatocarpa
(1972) D.kymatocarpa
D.kymatocarpa
D.pruinosa
D.pruinosa
D.pruinosa
D.inoxia
D.inoxia
D.inoxia
D.inoxia
D.metel
D.metel
D.metel
D.metel
D.metel
D.lanosa
D.lanosa
D.lanosa
D.lanosa
D.wrightii
D.wrightii
D.wrightii
0.00 0.08 0.15 0.22 0.30
Genetic distance

clustered together and related to D. inoxia and
D. wrightii, D. inoxia is divided between Clusters 2
and 3, and, within Cluster 2, it is further divided
among different branches. The weak relationship
among the populations labeled D. inoxia suggests
that this taxon is polyphyletic or that the identity of
the specimens is erroneous.
Our study used different isozymes and recognized
the polymorphism of loci in order to establish the
relationship among the taxa of section Dutra using
genetic distance. All taxa cluster among themselves
and the section is divided into two groups consisting
of D. inoxia, D. metel, D. wrightii, and D. lanosa in
one and D. discolor, D. kymatocarpa, D. pruinosa,
and D. reburra in the other. These groups were
suggested partially in the previous peroxidase and
AFLP studies.
In the phylogenetic analysis, the cladogram
revealed that the annuals form one clade while the
long-lived perennials are located in a separate clade.
In this latter clade, D. inoxia and D. metel are paired
and are related subsequently to D. wrightii and
D. lanosa. The D. inoxia–D. metel subclade is con-
sistent with the results of the previous phenetic
analyses of morphological and isozymatic characters
(Luna-Cavazos et al. 2000; Jiao et al. 2002). This
grouping also coincides with analyses using
cytogenetic, chemical, and hybridization data (Satina
1959; Palomino et al. 1988; Bye et al. 1991).
All eight species of the section Dutra grow
naturally in Mexico with four of these occurring as
Mexican endemics (Fig. 3). Datura metel is culti-
vated in various parts of the country but grows
commonly in the tropical lowlands where it is found
in gardens and escaped along the coasts of the Gulf of
Mexico and Caribbean Sea and across the Isthmus of
Tehuantepec to the Pacific Ocean. Datura inoxia has
a wide distribution in the tropical and temperate
uplands of Mexico. A phenetic analysis demonstrated
the greatest similarity between populations of D. me-
tel and those of D. inoxia in the southern Yucatan
Peninsula (Luna-Cavazos et al. 2000). At the oppo-
site end of the geographic range, D. wrightii is
common in southwestern United States with a few
populations extending into northwestern Mexico
where the other northern species, D. lanosa, is
restricted.
Based upon the phylogenetic analysis and its
corroboration in previous studies, we propose that
D. metel was derived from a common ancestor with
D. inoxia. Further evidence comes from the biogeo-
graphic distribution of D. metel in southern Mexico
and adjacent Caribbean area and Central America
where it is sympatric with that of D. inoxia in the

123
272 Genet Resour Crop Evol (2009) 56:263–275

from the other long-lived perennial members and

Fig. 2 Cladogram illustrating the phylogenetic relationship
among the species of Datura section Dutra and Brugmansia
sanguinea. The black rectangle indicates synapomorphy, the
parallel lines represent homoplasy, and the ‘‘X’’ is reversal.
The number on the left refers to the characters listed in Table 2
and that on the right indicates the changed character state. The
Consistency Index = 0.690. The support value is shown to the
side of its respective clade or subclade
southern part of its range. The northern allopatric
ranges of D. wrightii and D. lanosa do not place
them in geographic proximity of contemporary
D. metel. The D. inoxia–D. metel subclade diverged
underwent sympatric speciation.
Considering the evidence from morphological,
genetic, and eco-phytogeographical studies, one can
explain the continuum between a wild progenitor and
its cultivated domesticate (Hancock 1992; Harlan
1992). From the taxonomic view, the ancestral
species has characteristics similar to those of the
domesticate (Ohnishi 1998) taking into account the
distortion of certain desirable characters of the
human-selected plants. Such similarity between these
taxa led to misidentifications in earlier studies of
Datura (Satina and Avery 1959). The most obvious
differences between D. inoxia and D. metel are the
purple and white coloration patterns and the addi-
tional verticillate corollas of the ornamental D. metel
as well as its less threatening fruit with reduced
spines on the exocarp.
Genetically, the domesticated species should have
more alleles of common loci with its progenitor than
with other similar species; hence the multivariate
analysis of isozymatic data would group the domes-
ticated species closer to its wild progenitor (Doebley
1989). In the case of Datura, D. metel shares more
alleles with D. inoxia and D. lanosa than the other
species in the section Dutra. In a similar study of
Solanum tuberosum with 13 isozymatic loci of wild
and cultivated species of the Andean taxa (Oliver and
Martinez-Zapater 1984), S. stenotomum Juz. et Buka-
sov and S. sparsipilum (Bitter) Juz. et Bukasov were
identified as probable progenitors of the common
potato. The analysis of isozymatic data transformed
to genetic distance (McLeod et al. 1983)

Fig. 3 Geographic
distribution of the species of
Datura section Dutra. ,
Datura discolor; ,
D. inoxia; ,
D. kymatocarpa; ,
D. lanosa; , D. metel; ,
D. pruinosa; ,
D. reburra; ,
D. wrightii

123
Genet Resour Crop Evol (2009) 56:263–275
demonstrated that C. annuum L. var. baccatum is the
probable progenitor of the cultivated C. annuum var.
pendulum with the lowest genetic distance between
them (D = 0.02). In the multivariate analysis of
isozymatic data of Datura, the D. inoxia was most
similar to D. metel (D = 0.037) than the other
species.
Our cladistic analysis indicates that D. metel and
D. inoxia share the same ancestor but differ in external
floral characters, pubescence, and fruit ornamentation.
These divergences probably arose due to preferences for
desirable characteristics of interest to humans. Because
of its importance as an ornamental, the showier flower
of D. metel is the result of artificial selection of
ancestral D. inoxia-like populations for altered corollas
and diversified color patterns. In addition the longevity
of the attractive flower (up to one week) contrasts with
the ephemeral flowers of all the wild Datura species
which open at dusk and wilt the following morning.
Arresting flower senescence places an additional burden
on the plant in terms of floral maintenance and
construction cost (Ashman and Schoen 1994) that
increases its dependence upon humans for its survival.
Rare mutations that increase the number of corolla
verticils from one to two or three produce more
attractive flowers (Reynolds and Tampio 1983). Gla-
brous leaves and spineless fruits allow easier handling
of the plants rather than hairy and prickly herbs.
The phytogeographic distribution also supports the
phylogenetic pattern. Datura metel and D. inoxia
overlap in the southern range of the latter taxon. Such
sympatry is evidence used to identify progenitors of
Fagopyrum (Ohnishi and Matsuoka 1996; Ohnishi
1998). Parapatric and sympatric speciation modes are
important in plant domestication (Hancock 1992).
The cultivation of desirable plants with D. metel-like
characteristics in home gardens isolates the preferred
plants from its relatives in naturally disturbed habits
and provides an ecogeographic reproductive isolation
barrier. In addition the suppression of flower senes-
cence and reduction of reproductive parts in the
D. metel-like flowers could promote direct selection
on mating traits that are known to facilitate sympatric
speciation (Bolnick and Fitzpatrick 2007).
In conclusion, D. metel is most similar to D. inoxia
based upon morphological and isozymatic characters.
Cladistic analysis shows that D. metel and D. inoxia
are not only in the same subclade of the section Dutra
but are derived from an immediate common ancestor.
273
The phytogeography of section Dutra places D. in-
oxia as the only sympatric species with the geographic
distribution D. metel. The domesticated D. metel
arose most likely as a product of artificial selection
from wild D. inoxia-like populations in southeastern
Mexico, where they were favored for their attractive,
long-lasting flowers and less noxious fruits.
Acknowledgements We thank Alfredo Cervantes, Fernando
Chiang, Patricia Dávila, Alfonso Delgado, Patricia Escalante,
Victor Fuentes, Les Landrum, Gary Nabhan, Juan Nuñez-
Farfán, Porfirio Ramı́rez, David Spooner, Oswaldo Tellez, and
John Turrell for their suggestions and helpful reviews during
the development of this work. We acknowledge the
bibliographic, field, and technical assistance of José Arellano,
Jennifer Bain, Francisco Basurto, Bruce Bartholomew, Germán
Bojórquez, Lourdes Carmona, Rafael Corral, Carlos Dı́az,
Francisco Felix, Oscar Ferrera, Raymundo Garcı́a, Martı́n
Hilerio, Elia Herrera, Ma. Antonieta Isidro, Edelmira Linares,
Rigoberto López, Gilberto Márquez, Miguel Ángel Martı́nez,
Myrna Mendoza, Gustavo Morales, Eduardo Palacios, Isaac
Reyes, Lourdes Rico, Joel Rodrı́guez, Victoria Sosa, Richard
Spellenberg, Miguel Trejo, and Hugh Wilson. The main
electrophoresis works were carried out at the Laboratory of
Electrophoresis of Instituto de Biologı́a de la UNAM; special
thanks go to Fernando Cervantes and members of his group.
Nidia Pérez from the Instituto de Ecologı́a de la UNAM
provided initial guidance for electrophoresis techniques. Partial
financial support for this work was provided by Comisión
Nacional para el Conocimiento y Uso de la Biodiversidad
(Project 088), International Cooperative Biodiversity Groups
(‘‘Bioactive agents from dryland biodiversity of Latin
America’’ grant U01 TW 00316 from the National Institutes
of Health, National Science Foundation, and USAID), and
Universidad Nacional Autónoma de México. CONACYT
provided partial scholarship support to Mario Luna-Cavazos.
Dirección General de Asuntos del Personal Académico de la
Universidad Nacional Autónoma de México provided a partial
scholarship to Meijun Jiao.
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