Annals of Botany 96: 887–900, 2005

doi:10.1093/aob/mci241, available online at www.aob.oxfordjournals.org

A Morphological Study of the Petunia integrifolia Complex (Solanaceae)

T O S H I O A N D O 1,*, N O B U Y U K I I S H I K A W A 1, H I T O S H I W A T A N A B E 2, H I S A S H I K O K U B U N 3,
Y O S H I K I Y A N A G I S A W A 3, G O R O H A S H I M O T O 4, E D U A R D O M A R C H E S I 5 and
E N R I Q U E S U Á R E Z 6
1
Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo City, Chiba 271-8510, Japan, 2Center of Environment,
Health and Field Science, Chiba University, 6-2-1 Kashiwanoha, Kashiwa City, Chiba 277-0882, Japan, 3Graduate School of

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Science and Technology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba City, Chiba 263-8522, Japan, 4Centro de
Pesquisas de História Natural, 618 Rua Jaime Ribeiro Wright, Itaquera, São Paulo 08260-070, Brazil, 5Facultad de
Agronomia, Universidad de la República, Garzón 780, Montevideo, Uruguay and 6Centro de Investigation de Recursos
Naturales, INTA, Las Cabañas y Reseros s/n (1712), Castelar, Prov. de Buenos Aires, Argentina

Received: 23 January 2005 Returned for revision: 4 April 2005 Accepted: 20 June 2005 Published electronically: 15 August 2005

 Background and Aims Petunia inflata has been treated taxonomically in various ways: it has been described as an
independent species, treated as a synonym of P. integrifolia, and also regarded as a subspecies of P. integrifolia. The
present study was designed to resolve the ambiguity involving the P. integrifolia complex (P. integrifolia plus
P. inflata).
 Methods Tentative identification (either integrifolia group or inflata group) was carried out in the field based on the
observation of live specimens at the restricted type localities. The accuracy of the tentative identification was later
tested with principal component and cluster analyses of data obtained by measuring 21 morphological characters on
cultivated live specimens sourced from 113 natural populations of the P. integrifolia complex in Argentina, Brazil,
Paraguay and Uruguay.
 Key Results There was a clear, statistically significant gap between the morphological measurements of the two
groups, ensuring the accuracy of identification carried out in the field except for a probable hybrid swarm.
Previously, the condition of the pedicel in the fruiting state was considered an important character distinguishing
between these two groups; however, the condition of the pedicel was rather variable in the integrifolia group. The
two groups were found to have geographically distinct distributions: the integrifolia group occurred in southern
regions, whereas the inflata group occurred in northern regions.
 Conclusions Based on the available evidence, it is suggested that the two groups are allopatric species,
P. integrifolia and P. inflata, in agreement with the opinion of Fries (1911).

Key words: Distribution, floral morphology, Petunia, Serra Geral, Solanaceae, South America, speciation.

INTRODUCTION several European herbaria and reported inter-gradation in

pedicel condition and floral size between the three species
The first garden petunia (Petunia · hybrida Vilm.,
over a geographical course, moving west from southern
Solanaceae) was reported to be an interspecific hybrid
Brazil (P. integrifolia) through Paraguay and north-
of P. axillaris (Lam.) Britton, Sterns & Poggenb.
eastern Argentina (P. inflata) to north-western Argentina
(=P. nyctaginiflora Juss.) and P. integrifolia (Hook.) Schinz
(P. occidentalis). In that study, the deflexus characters
& Thell. (=P. violacea Lindl.) (Paxton, 1836). Since then,
were reported to change to inflexus, and to coincide with
Petunia inflata R. E. Fr. and P. parodii Steere [=P. axillaris
a reduction in flower size along the geographical cline.
subsp. parodii (Steere) Cabrera] have also been proposed as
Wijsman (1982) provided several subspecies names, such
the likely parents of modern garden petunias (Sink, 1984).
as subsp. integrifolia, subsp. inflata and subsp. occidentalis,
Within these proposed ancestors, the taxonomic position of
for the ‘extremes’ of P. integrifolia. However, a previous
P. inflata has not yet been clearly defined. Fries (1911)
study comparing morphology among his infraspecific taxa
originally described P. inflata as an independent species
of P. integrifolia, as above, failed to confirm the inter-
based on a comparison of morphological characters with
gradation reported by Wijsman (1982); instead, distinct
P. integrifolia. Fries (1911) noted that one of the main
morphological differences were found between the taxa
features distinguishing these two taxa is the pedicel condi-
concerned (Ando et al., 1995). Based on these studies and
tion in the fruiting state. In P. integrifolia, the pedicels are
other evidence obtained from a cross-compatibility study,
deflexed, whereas in P. inflata they are inflexed. Sub-
P. occidentalis was resurrected as an independent species
sequently, however, Smith and Downs (1966) regarded
(Tsukamoto et al., 1998). At that time, however, there was
P. inflata as a synonym of P. integrifolia without providing
a hesitancy to come to a conclusion regarding the taxonomic
any clear reason. Wijsman (1982) later studied specimens
positions of subsp. integrifolia and subsp. inflata because
of P. integrifolia, P. inflata and P. occidentalis R. E. Fr. in
earlier experiments were conducted on a small scale.
* For correspondence. E-mail andot@faculty.chiba-u.jp Accordingly, Wijsman’s P. integrifolia subsp. integrifolia

The Author 2005. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.
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888 Ando et al. — Petunia integrifolia Complex
and subsp. inflata, which the authors describe as the
‘P. integrifolia complex’, were left for subsequent detailed
studies.
Since Wijsman’s (1982) work, Ando and Hashimoto
(1993, 1994, 1995, 1996, 1998) described several new
species of Petunia from uplands of southern Brazil based
on the observation of native live plants. All the new species
are unique in floral and other morphology and are readily
distinguishable from previously known species includ-
ing ‘P. integrifolia complex’. Fries (1911) recognized a
subspecies of P. integrifolia (subsp. depauperata),
which occurred along the Atlantic coast of Brazil. Smith
and Downs (1966) treated this taxon as a variety of
P. integrifolia, while Wijsman (1982) regarded this as a
variant growing in nutrient-poor soil. However, this plant
produces extremely long prostrated stems that bear sparse
small flowers and linear leaves, and such features in mor-
phology are never found in any taxa of Petunia with the
exception of P. littoralis L. B. Sm. & Downs. A comparison
of the morphology and the distribution range of this taxon
with those of P. integrifolia will be reported elsewhere.
The objective of the present study was to evaluate
Wijsman’s Petunia integrifolia subsp. integrifolia and
subsp. inflata using multivariate analyses of morphological
characters. The first author has travelled >140 000 km in
temperate and subtropical South America, and observed
448 natural populations of the P. integrifolia complex
over a substantial part of the distribution range. In addition
to being a candidate for the ancestor of garden petunias,
as mentioned above, P. inflata is also an important experi-
mental model system for studies of solanaceous self-
incompatibility (Dowd et al., 2000). Therefore, there is a
clear need to resolve some of the taxonomic ambiguity
involving this taxon.
MATERIALS AND METHODS
Observation at the restricted type locality
Hooker (1831) described Petunia integrifolia (=Salpiglossis
integrifolia) based on two specimens. The source of one of
the specimens was uncertain, while J. Baird collected the
other near the Rı́o Negro, Uruguay. In order to observe live
specimens of P. integrifolia at this location, the first author
visited the Rı́o Negro, Uruguay (on 28 Nov. 1989), and
adjacent regions (Ando, 2003, 2004). For Petunia inflata,
Fries (1911) established several syntypes. The first author
also visited a number of locations in Argentina, including
Bonpland in Misiones Province (on 27 Nov. 1990), one of
the restricted type localities, and adjacent regions (Ando,
2003, 2004).
Plant materials
Seeds and herbarium specimens were collected between
1988 and 1999. Based on observations of live plants
at the above locations, tentative identification of the
material as either P. integrifolia or P. inflata was carried
out for the native populations. Seeds of P. integrifolia
(57 populations) and P. inflata (56) were collected over a
substantial distribution range of the P. integrifolia complex.
Additional samples were collected from Rio Grande do Sul
(RS) in Brazil, since the distributions of the two taxa are
very similar in that region.
The following notation was used for the accession codes
representing the source of the collected samples: ‘A’,
Argentina, ‘B’, Brazil, ‘P’, Paraguay, and ‘U’, Uruguay
(see the Appendix). The location of each population was
recorded using a global positioning system (GPS) receiver
[Sony IPS/360 (Sony Co., Ltd, Shinagawa, Tokyo, Japan),
Panasonic KX-G5550 (Matsushita Electric Industrial Co.,
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Ltd., Kadoma, Osaka, Japan) or GPS III Plus (Garmin,
Olathe, KS, USA)].
For all but 12 of the populations [B272, B522, B692,
B724 (nine individuals), B688, B841, U128 (8), B809,
U12 (7), B1206, U168 (6) and A166 (5)], ten individuals
were raised from seed in a greenhouse, following the stand-
ard procedure for cultivating garden petunias. The plants
were grown in 15-cm (diameter) pots, and the lateral stems
were held vertical using plastic supports.
Measuring live plants
In a previous study, 33 morphological characters encom-
passing reproductive and other traits (X1–X33) were com-
pared in each of the infraspecific taxa of Petunia integrifolia
sensu Wijsman (1982) (Ando et al., 1995). For the present
study, 21 of the morphological characters that had been
found at that time to be useful for distinguishing subsp.
integrifolia and subsp. inflata were chosen. The code
numbers used for the characters follow those of Ando
et al. (1995) (Table 1).
The morphological features were measured on all of the
greenhouse-grown specimens between May and July 2001.
One typical flower was chosen per plant and a mean value
was calculated for each population. Characters were
measured using digital calipers (Mitutoyo Co., Kawasaki,
Kanagawa, Japan) when the plants reached full bloom. To
measure the pedicel angle in the fruiting state and other
morphological characteristics related to the capsule, selec-
ted flowers for each plant were crossed with pollen from a
different individual belonging to the same population. The
angle of the pedicel was measured when the capsule reached
maturity.
Measuring herbarium specimens
Pedicel angles were also measured on voucher herbarium
specimens collected from each of the native populations. Up
to five pedicels with mature capsules were selected for the
measurement. A capsule was considered mature when it
dehisced or reached full size. The ratio of mature capsules
with folded calyx lobes to those with straight (not folded)
ones was also recorded for the voucher specimens. The first
author visited four European herbaria [Herbarium, Botany
Department, Natural History Museum, London (BM); Herb-
arium, Royal Botanic Gardens, Kew (K); Rijksherbarium,
Leiden (L); and Herbarium, Institute of Systematic Botany,
State of Utrecht, Utrecht (U)] to re-inspect the specimens
that Wijsman (1982) studied.
 Ando et al. — Petunia integrifolia Complex 889
T A B L E 1. Morphological and other characters measured in the study: the means and results of the principal component analysis

PC analysis

Characters inflata integrifolia Factor loading

Code† Definition Mean CVz Mean CVz 1st PC 2nd PC 3rd PC 4th PC

X1 Distance between anthers of long and 3·50 8·2 4·00 6·7***x 0·792 0·045 0·100 0·060
medium stamens (mm)
X3 Distance between two anthers of 2·49 5·8 2·62 8·9*** 0·424 0·255 0·770 0·085
long stamens (mm)
X4 Distance between two anthers of 2·40 7·4 2·43 9·1 n.s. 0·261 0·553 0·656 0·127

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medium stamens (mm)
X5 Length of long stamens (mm) 14·87 3·9 18·22 5·2*** 0·947 0·193 0·059 0·031
X6 Length of basal part of filament 4·52 7·9 7·27 10·3*** 0·941 0·246 0·008 0·108
affixed to corolla-tube (mm)
X7 Proportion of X6 to X5: (X6/X5) · 100 (%) 30·44 7·5 39·84 6·7*** 0·871 0·291 0·065 0·148
X8 Length of pistil (including ovary) (mm) 12·94 3·9 16·00 5·8*** 0·940 0·205 0·020 0·046
X9 Length along vertical axis of stigma (mm) 0·91 11·4 1·39 10·0*** 0·918 0·195 0·073 0·030
X10 Length along horizontal axis of 1·21 13·3 1·48 9·6*** 0·806 0·293 0·090 0·003
stigma (mm)
X11 Ratio X9 : X10 0·77 17·2 0·95 9·3*** 0·527 0·559 0·181 0·045
X12 Length of upper calyx lobe (mm) 11·86 12·4 13·52 13·8*** 0·577 0·134 0·038 0·512
X14 Length of calyx tube (mm) 2·40 12·5 2·87 10·1*** 0·581 0·052 0·020 0·146
X16 Length along long axis of ovary (mm) 2·32 7·2 2·51 6·7*** 0·653 0·395 0·162 0·390
X17 Length along horizontal axis of ovary (mm) 1·39 5·7 1·49 5·4*** 0·679 0·343 0·161 0·325
X19 Ratio X6 : X16 1·98 9·9 2·93 10·6*** 0·833 0·424 0·059 0·256
X21 Length of corolla tube (mm) 21·55 5·1 24·90 7·0*** 0·919 0·028 0·115 0·001
X24 Diameter of corolla tube (mm) 7·70 6·1 10·00 5·7*** 0·910 0·093 0·012 0·037
X25 Length of corolla (mm) 31·36 6·1 35·96 8·1*** 0·874 0·117 0·149 0·061
X30 Length of capsule (mm) 5·99 11·1 6·34 9·7** 0·484 0·614 0·230 0·374
X31 Width of capsule (mm) 3·57 8·5 3·87 11·1*** 0·567 0·394 0·326 0·479
X33 Angle of pedicel to stem during fruiting ( ) 74·22 11·9 131·09 30·6*** – – – –
Eigenvalue 11·31 2·06 1·34 1·05
Variance (%) 56·6 10·3 6·7 5·2
Cumulative 56·6 66·9 73·6 78·8
variance (%)

†
After Ando et al. (1995).
z
Coefficient of variation (%).
x
Level of significance by t-test at: ***, 0·1 %; **, 1·0 %.

Multivariate analysis used squared Euclidian distances, and the amalgamation

Fries (1911) studied herbarium specimens and regarded
the pedicel condition in the fruiting stage as a con-
stantly reliable key morphological character distinguishing
P. inflata (inflexed) from P. integrifolia (deflexed). There-
fore, character X33 (angle of pedicel to stem during fruiting)
was not included in the multivariate analysis, in an attempt
to find other characters differentiating the taxa.
The resulting data matrix contained 20 variables (means
of the measurements for 20 morphological characters)
and 113 cases (populations). This dataset was used for a
principal component (PC) analysis to separate populations
based on measured morphological characters, and to
identify the morphological characters that differed between
the groups. The PC analysis was carried out using the
FACTOR procedure in the SPSS statistical package
(SPSS Inc., Chicago, IL, USA).
Cluster analysis was carried out using a similar data
matrix, except that it lacked the B812 population (discussed
below). The PROXIMITY and CLUSTER procedures in
SPSS were used to calculate a distance matrix and an
amalgamation schedule, respectively. The distance matrix
method used the un-weighted pair-group method on the
arithmetic mean (UPGMA). The FACTOR procedure
was also used on this data to obtain correlation coefficient
(CC) matrices among variables for the inflata and integri-
folia groups separately. The T-TEST procedure in SPSS
was used to compare means between the two groups and
between two subclusters of the integrifolia group.
Distribution map
A distribution map was produced using TNTlite
(MicroImages Inc., Lincoln, NB, USA). Political boundar-
ies and rivers were taken from the Digital Chart of the
World (ESRI Inc., Redlands, CA, USA). The Rio Icamaquã
and Rio Vacacaı́, which were added to the map manually,
were sourced from a 1:250 000 topographical map (Instituto
Brasileiro de Geografia e Estatı́stica, Rio de Janeiro, Rio de
Janeiro State, Brazil). Contour intervals (200 m) were gen-
erated from GTOPO30 DEM data (USGS, Reston, VA,
USA). Collection localities for each of the populations
were overlaid as a database pinmap, using GPS co-ordinates
taken during the field survey.
890 Ando et al. — Petunia integrifolia Complex
3 B560
B1270
PC2 B522
B1222
B377
2 inflata group B1163
B1202
B812 B574
B1219
B564
1 B581
B531
B1302
B956
B261
0 P1
PC1 B719
B723
B350
B439
–1 B628
B724

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B587
B954
–2 integrifolia group B1203
B1204
B1212
A249
A260
–3 B533
B698
–2 –1 0 1 2 3 B556
B1206
B1209
F I G . 1. Two-dimensional scatter diagram of the first and second PC scores B694
of floral morphology of the integrifolia group (circles), the inflata group B709
B549
(triangles), and an intermediate form (solid square). Character X33 was B697
B692
omitted from the calculation. B274
B538
B540
A7
B269
RESULTS B1200
A270
B1208
PC and cluster analyses B1210
B710
B1287
When the PC analysis was performed on the data matrix A119
B1288
consisting of 20 variables (morphological characters other B272
B1211 inflata group
than X33) and 113 cases (populations), four PCs with eigen- B544
B1205
values larger than 1·0 were obtained. These four groups B818
B848
explained 78·8 % of the total variance in the data matrix B817 integrifolia group
B1192
B1198
(Table 1). A two-dimensional scatter diagram for the first B314
B677
and second PC scores for the respective populations showed U128
B294
that two groups occupied separate regions, and one popu- B311
B687
lation (square, B812 population) was intermediate (Fig. 1). B1195
B301
The restricted type localities of P. integrifolia (U106, B1314
B300
U168 and U239) were included in a group that was distrib- A163
U12
uted mainly in the first and fourth quadrants of Fig. 1 (open A168
B704
and closed circles, 56 populations). The plants grown from B948
B802
all populations of this group were identified in the field as B673
B702
belonging to P. integrifolia. Hereafter, plants from these B688
B701
populations are referred to as the integrifolia group. The U168
B809
A171
restricted type localities of P. inflata (A7, A119 and P1) U106
A175
belonged to another group distributed in the second and U239
U263
third quadrants (triangles, 56 populations). Specimens U231
B705
from all populations of this group were identified in the A166
B680
field as belonging to P. inflata. Hereafter, these populations U46
U161
are referred to as the inflata group. B682
B868
Plants from the B812 population were identified in the B315
B867
field as P. integrifolia. When data obtained from ten indi- B651
U233
viduals from the B812 population were added to the original B280
B1190
data matrix and subjected to PC analysis, two and three B1304
B1193
individuals were distributed in the integrifolia and inflata B686
B708
B707
groups in the scatter diagram, respectively. The remaining •B835
•B841
five individuals fell into the region intermediate between the •B870
•B949
two major groups (data not shown). In an attempt to clarify •B971
the morphological differences between the inflata and
integrifolia groups, the B812 population was removed 0 30 60
from subsequent analyses because it was considered a Amalgamation distance
hybrid swarm. F I G . 2. Cluster analysis using 20 characters representative of reproductive
The dendrogram produced from the modified (no B812 organ morphology of the integrifolia and inflata groups. Character X33 and
population) data matrix showed two distinct clusters (Fig. 2). B812 population were omitted from the calculation.
 Ando et al. — Petunia integrifolia Complex 891

A X6 B X6

X16 X16

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F I G . 3. Longitudinal section of flowers indicating two measurements (X6, length of basal part of filament affixed to corolla-tube; X16, length along long axis
of ovary): (A) the integrifolia group (U106 population); (B) the inflata group (A7 population). Two calyx lobes were eliminated. Scale bar = 10 mm.

T A B L E 2. Correlation coefficients between the length of the corolla tube (X21)/total corolla (X25) and the length of the inner
floral organs

Length of corolla tube (X21) and total corolla (X25)

integrifolia group inflata group

Length of X21 X25 X21 X25

long stamen (X5) 0·8633***† 0·7489***† 0·5118***† 0·3925**

basal part of filament affixed to the corolla tube (X6) 0·7657*** 0·6257*** 0·1243 n.s. 0·1331 n.s.
pistil (X8) 0·8446*** 0·7688*** 0·3400* 0·2617 n.s.
along long axis of ovary (X16) 0·4092** 0·4213** 0·6546*** 0·6659***

†
Level of significance at: ***, 0·1 % level; **, 1·0 % level; *, 5·0 % level.

The upper cluster of the dendrogram consisted of members X6 < 5·23 mm, and length along vertical axis of stigma
of the inflata group and the lower cluster was made up of X9 < 1·12 mm) from members of the integrifolia group
members of the integrifolia group. (X6 > 5·78 mm, and X9 > 1·39 mm) (Fig. 3).

Correlation between flower size and length of the inner

Differences in the morphological characters
floral organs
As assessed by the larger absolute values of the factor
Table 2 shows the correlation coefficients (CCs) between
loading, characters related to the stamens (X5, X6 and their
two characters related to flower size (X21, length of corolla
proportion X7), pistil (X8), stigma (X9 and X10), corolla
tube; and X25, length of total corolla) and four characters
(X21, X24 and X25), and X19 (the ratio of X6 : X16) were
related to the size of the inner floral organs (X5, length of
selected as the morphological characters that were most
long stamen; X6, length of basal part of filament affixed to
differentiated between members of the integrifolia and
corolla tube; X8, length of pistil; and X16, length of ovary).
inflata groups (Table 1, shown in bold face). The mean
In the integrifolia group, the size of the inner floral organs
values obtained for the morphological characters of the
(X5, X6, X8 and X16) resulted in higher CCs (1·0 % signi-
inflata group were significantly smaller than those of the
ficant level) compared with characters related to flower size
integrifolia group, with the exception of character X4. This
(X21 and X25). For the inflata group, character X6 did not
result was mostly confirmed with a t-test at the 0·1 % level
correlate with characters X21 or X25, and character X8 did
of significance (Table 1), and was in agreement with a
not correlate with character X25.
previous study conducted on a smaller scale (Ando et al.,
1995). The ranges of measurements of the inflata group for
Identification in the field
three characters (length of long stamens X5 < 16·42 mm,
length of pistil X8 < 14·1 mm, and diameter of the corolla Although a subset of morphological characters of the
tube X24 < 8·5 mm) were clearly separate from those of inflata group differed from those of the integrifolia group,
the integrifolia group (X5 > 16·44 mm, X8 > 14·5 mm, and as described above, field identification was based mainly
X24 > 9·0 mm). Considering the 95 % range (61·96 · on other morphological characters. Slight differences in the
standard deviation), two more characters were also found colour of the flower were one of the characters used for
to be useful for separating members of the inflata group identification. The flowers of both taxa have been described
(length of basal part of filament affixed to corolla tube as reddish-purple, but in the authors’ experience, the tint of
892 Ando et al. — Petunia integrifolia Complex
the inflata group is slightly brighter than that of the A
integrifolia group. This difference is evident from a distance inflata group
of several metres, especially on cloudy days. integrifolia group
A significant difference in the diameter of the corolla
tube (X24) (Table 1) was also apparent on visual inspection
of the flower. The mouth of the corolla was larger in the

Frequency
integrifolia group than in the inflata group (see fig. 5 in 10
Ando et al., 1995).
The colour and venation pattern on the inner-surface
of the corolla tube was also a character that could reliably
distinguish the inflata group from the integrifolia group. The

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colour of the inside of the corolla tube was much paler than
the corolla limb in the inflata group, while in the integrifolia
group the corolla tube was darker than the corolla limb. In
0
the inflata group, darker coloured veins were restricted to a 40 65 90 115 140 165 190 215
small area on the upper side of the inner corolla tube
(Fig. 3B). The pattern of venation expanded widely inside X33: angle of pedicel to stem during
fruiting (degrees) in live plants
the corolla tube in the integrifolia group, although this was
not clearly visible, due to the darker ground colour (Fig. 3A).
Using these characters, it was possible to distinguish indi- B inflata group
viduals belonging to the inflata and integrifolia groups in integrifolia group
the field.

Pedicel condition
Frequency

Character X33 (angle of the pedicel relative to the stem
during fruiting) was measured on plants cultivated from
seed. For the inflata group, angles between 53 and 91
(mean 74 ) were obtained. For the integrifolia group, the
angle varied between 45 and 218 (mean 131 ) (Fig. 4A).
Among the herbarium specimens belonging to the inflata
group, the range was much smaller (31–77 ; mean 53 )
(Fig. 4B). The pedicel angle of the integrifolia group recor-
ded from herbarium specimens showed a narrower range
(48–150 ; mean 108 ) compared with cultivated plants. The
typical features of the pedicel characters are shown in
Fig. 5A (integrifolia group) and B (inflata group).
Another character observable in herbarium specimens
The calyx lobes located next to the maturing capsules of
specimens belonging to the integrifolia group were patent
and out-curved, whereas those of the inflata group were
closed and straight. This subtle difference changed the
appearance of the calyx lobes, and was easily observed
in the herbarium specimens (Fig. 5A and B). In most
cases, at least one of the five calyx lobes of the integrifolia
group was folded (see arrows, Fig. 5A). By contrast, all the
calyx lobes of the inflata group were usually straight
(Fig. 5B). The frequency distribution of the mean values
is shown in Fig. 6.
Geographic distribution
Figure 7 shows a topographical map (200-m intervals) of
the study region, indicating the location of populations
of the integrifolia (open and solid circles) and inflata
(triangles) groups, and population B812 (solid square).
The distribution range of the inflata group clearly differed
from that of the integrifolia group. The inflata group
was located in northern areas, whereas the integrifolia
group was located in southern areas. In Brazil, the inflata
10
0
40 65 90 115 140 165 190 215
Angle of pedicel to stem during fruiting (degrees)
in herbarium specimen
F I G . 4. Frequency distributions of mean pedicel angle: (A) measured from
cultivated live plants; (B) measured from herbarium specimens.
group was found in upland areas in northern RS and Santa
Catarina (SC) states, whereas the integrifolia group was
found in lowland areas, such as along the Rio Ibicuı́ flow-
ing west, and the Rio Jacuı́ and its upper stream (the Rio
Vacacaı́), which both flow east (Fig. 7).
Outside Brazil, the inflata group was the sole taxa found
in the Chaco, Corrientes, Formosa and Misiones Provinces
of Argentina, and Paraguay while the integrifolia group was
found exclusively in the Entre Rı́os Province of Argentina,
and Uruguay. Compared with Brazil, the range of the inflata
group extended much further north in Paraguay and Argen-
tina, up to 25 S (Fig. 7). In Corrientes Province, the range of
the inflata group was restricted to the northern region, close
to Paraguay. The range of the integrifolia group was con-
centrated along major rivers, such as the Rı́o Uruguay and
Rı́o Negro (Fig. 7).
Sub-clusters of the integrifolia group
The dendrogram (Fig. 2) shows that amalgamation
distances were smaller in the inflata group than in the
 Ando et al. — Petunia integrifolia Complex 893

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A B
F I G . 5. Typical features of the pedicel and calyx lobes observed in herbarium specimens: (A) integrifolia group (B688); (B) inflata group (B1206).
Scale bars = 50 mm.

50 this was not necessarily the case for members of the minor
inflata group sub-cluster (Fig. 7, solid circles). Populations belonging to
integrifolia group the minor sub-cluster were often located on steep sloping
areas and ascending uplands (B870 and B949) and adjacent
lowland (B835 and B971) in central RS and upland areas in
south-western RS (B841) (Fig. 7).
Frequency

Specimens inspected by Wijsman (1982)

10
The first author re-inspected two of the three specimens
of the P. integrifolia complex that exhibited a pedicel
condition described by Wijsman (1982) as ‘deflexed, but
some aberrant’. The first specimen was found to be neither
a member of the integrifolia nor inflata groups, but was a
recently described species, Petunia altiplana T. Ando &
Hashim. (Lindeman & de Haas 3621 deposited in U).
0 The second specimen was not a species of Petunia, but
0 25 50 75 100 was found to be a member of a sister genus, Calibrachoa
Rate of capsules with folded calyx linoides Sendtn. (Lindeman & de Haas 2649a in U). The
in herbarium specimen (%) third specimen, reported to have been collected from São
Joaquim in SC, Brazil (Reitz s. n. in U) could not be found.
F I G . 6. Frequency distribution of the percentage of capsules with folded
calyx lobes in herbarium specimens.

integrifolia group. In the integrifolia cluster, a minor sub-
cluster composed of five populations (B835, B841, B870,
B949 and B971) was recognized as an outlier (Fig. 2, solid
circles). Compared with the major sub-cluster, members
of the minor sub-cluster were characterized by significantly
larger flowers (X21 and X25), internal floral organs (X5, X6,
X8, X9 and X10) and capsules (X30 and X31). This trend to
larger organs was also evident for some other characters,
such as X7 and X19 (Table 3). In addition, members of the
minor sub-cluster had pedicel angles during fruiting (X34)
of around 90 (Table 3).
Although members of the major sub-cluster of the
integrifolia group always occurred in lowland habitats,
DISCUSSION
Identification of live plants
Although the taxonomic treatment of the integrifolia and
inflata groups is complicated as mentioned in the Introduc-
tion, it was possible to distinguish the two groups in their
natural habitats. The accuracy of the authors’ tentative
identification, based mainly on overall flower shape, colour
and venation patterns, was validated by the clear statistical
separation of two groups using PC and cluster analyses
(Figs 1 and 2), with the exception of a probable hybrid
swarm (B812 population).
Aside from the character of pedicel condition in the
fruiting state, which was not included in the multivariate
894 Ando et al. — Petunia integrifolia Complex

58°W 56°W 54°W 52°W 50°W

60°W 48°W
Formosa
Paraná

s
ione
26°S 26°S

Mi s
PARAGUAY m
200 0m
40 0m
100
Chaco

tarina
0m

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20

Santa Ca
28°S 28°S

10
60

80
0m

00
0m
400 m

m
São Borja

40
Corrientes

0m
200 m
ARGENTINA
30°S 30°S

l
Su
do
0m

20

de
0m
20

BRAZIL

ran
32°S Entre Ríos oG 32°S
Ri
0m
20

Atlantic Ocean
URUGUAY
34°S 34°S

58°W 56°W 54°W 52°W 50°W

60°W 48°W

F I G . 7. Geographical distribution of the integrifolia group (circles), the inflata group (triangles), and a possible hybrid swarm (solid square, B812).

analyses (Figs 1 and 2), several morphological measure-
ments distinguished members of the inflata and integrifolia
groups. The inflata group was characterized by significantly
shorter (or smaller) inner floral organs, such as stamens
(X5), basal part of the filament fixed to the corolla tube
(X6), pistil (X8), ovary (X16), calyx lobe (X12), calyx tube
(X14) and stigma (X9 and X10) (Table 1). The ranges of three
characters (X5, X8, and X24) for the inflata group did not
overlap those of the integrifolia group.
In the integrifolia group, the size of the inner floral organs
(X5, X6, X8 and X16) produced higher CCs with flower size
[as assessed by the length of the corolla (X25) or corolla tube
(X21)]. In the inflata group, the length of the basal part
of the filament affixed to the corolla-tube (X6) and the length
of pistil (X8) did not exhibit these features (Table 2),
suggesting that characters X6 and X8 are stable in the inflata
group irrespective of the flower size, which can vary
between populations in different local environmental con-
ditions. It is also noteworthy that two secondary characters,
X7 [proportion of X6 to the length of long stamen (X5)] and
X19 [ratio of X6 : the length of ovary (X16)], which are not
directly related to flower size, also differed significantly
between the inflata and integrifolia groups (Table 1).
The difference in character X19 showed that the basal
part of the filament affixed to the corolla tube (X6) is
1·98 (approx. 2) and 2·93 (approx. 3) times longer than
the length of the ovary (X16) in the inflata and integrifolia
groups, respectively (Table 1 and Fig. 3). From experience,
it is suggested that character X19 is one of the most con-
venient floral characters for distinguishing the inflata and
 Ando et al. — Petunia integrifolia Complex 895
T A B L E 3. Difference in the characters between major and minor sub-clusters of integrifolia group defined by cluster analysis

Mean Mean

Characters Major Minor units Clusters Characters Major Minor units Clusters

X1 3·97 4·22 mm n.s.† X14 2·91 2·45 mm n.s.†

X3 2·61 2·78 mm n.s. X16 2·50 2·61 mm n.s.
X4 2·39 2·79 mm * X17 1·49 1·54 mm n.s.
X5 18·03 20·07 mm ** X19 2·89 3·33 **
X6 7·13 8·62 mm ** X21 24·54 28·61 mm **
X7 39·53 43·00 % * X24 9·98 10·20 mm n.s.
X8 15·82 17·80 mm * X25 35·39 41·67 mm **

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X9 1·38 1·57 mm ** X30 6·26 7·17 mm ***
X10 1·46 1·74 mm * X31 3·81 4·45 mm *
X11 0·95 0·91 n.s. X33 134·97 91·48 degrees *
X12 13·38 14·94 mm n.s.

†
Level of significance by t-test at: ***, 0·1 %; **, 1·0 %; *, 5·0 %.

integrifolia groups. Unfortunately, many of the characters
that differentiate the two groups are fragile and are not
conserved in herbarium specimens.
Identification in the herbaria
In previous studies, Fries (1911) and Wijsman (1982)
highlighted the pedicel condition in the fruiting state as
the primary feature distinguishing the inflata and integrifo-
lia groups. By contrast, it was found that this character (X33)
is variable in the integrifolia group, as indicated by the large
coefficient of variation (CV) value (Table 1). In fact, char-
acters related to pedicel condition measured from cultivated
members of the integrifolia group (Fig. 4A) differed mark-
edly from those measured on voucher herbarium specimens
(Fig. 4B). Lateral stems of both subspecies are usually
ascending under natural conditions, but in the present
study, lateral stems were maintained in a vertical position
with the aid of plastic supports. Perhaps the manipulation of
the lateral stems caused this discrepancy.
For herbarium specimens, the pedicel angle for the inflata
group fell into a rather narrow range of <80 (Fig. 4B).
Therefore, it is reasonable to conclude that the inflata
group has inflexed pedicels at the fruiting stage. The pedicel
angle for integrifolia was more variable and did not always
exceed 90 (Fig. 4B). As a result, these cannot be considered
as reliable taxonomic features. A member of the Petunia
integrifolia complex can be identified as belonging to the
integrifolia group when it has deflexed pedicels, since this
characteristic does not occur in the inflata group (Fig. 4B).
If a specimen has a slightly inflexed pedicel, however, other
characters must be considered carefully before a clear
identification can be made.
In this study, another morphological character was found
that is readily observable in herbarium specimens and can
be used to distinguish the two groups. For the integrifolia
group at least one of the five calyx lobes is usually folded
(Fig. 5A), whereas in the inflata group, all the calyx lobes
are straight (Fig. 5B). It is worth noting that this character
is stable in the integrifolia group, but more variable in the
inflata group (Fig. 6), contrary to pedicel condition, as
mentioned above (Fig. 4). A herbarium specimen of the
Petunia integrifolia complex can be identified as belonging
to the inflata group when most of the capsules have straight
calyx lobes without folding, since this feature is rarely
observed in the integrifolia group (Fig. 6).
The identification of the P. integrifolia complex, as
described above, can be applied to mature plants; however,
it is often much more difficult to identify younger plants and
plants grown in shade. For mature plants, another index
character can be used. The upper leaves of mature plants
belonging to the inflata group tend to be very small and
scale-like (see the stem on the right-hand side of Fig. 5B).
This feature does not occur in the integrifolia group
(Fig. 5A). Based on this morphological feature, the inflata
and integrifolia groups can be distinguished without requir-
ing the inspection of inner floral organs.
Distribution
In Brazil, the integrifolia and inflata groups were isolated
geographically (Fig. 7). The inflata group occurred in north-
ern upland areas, on a lava plateau called the Planalto. This
region is characterized by rather flat, gentle hilly ground.
The steep slope at the edge of Planalto is known as the Serra
Geral. The elevation of the Planalto exceeds 1000 m at the
eastern end close to the Atlantic coast, and the Serra Geral
facing the east forms a perpendicular cliff. To the west, the
elevation of the Planalto gradually decreases, and the Serra
Geral facing south is strongly eroded to form a complex
terrain. The limit between the Planalto and the lowland area,
i.e. the border between the territories of the inflata and
integrifolia groups, is evident in central RS, but becomes
obscure in western RS. No geographical border, limiting the
territories of the inflata and integrifolia groups seems to
exist near the town of São Borja (indicated with a star in
Fig. 7), although the territories of the two groups are close to
each other at that point.
The region surrounding São Borja was visited four times
(1993, 1994, 1996 and 1997) to study distribution (Ando,
2003, 2004). The smallest distance between populations of
the integrifolia (B701, Mun. São Borja) and inflata (B699,
Mun. São Borja, route BR285, 5 km west-south-west of Rio
Icamaquã to São Borja, 28 570 4700 S, 55 310 4400 W) groups
896 Ando et al. — Petunia integrifolia Complex
was 16 km. Population B812, which was considered to be a
possible hybrid swarm, was situated within the territory of
the integrifolia group (solid square in Fig. 7), but only 30 km
from a population of the inflata group (B698, Mun. Santo
Antônio das Missões). The Rio Icamaquã, which flows close
to São Borja, appears as another border on the map (Fig. 7),
although this river seems too small to function as an obs-
tacle capable of physically separating the two groups.
Most of the research in this study was conducted in
Brazil, and the number of populations assessed from other
inland locations, such as Argentina and Paraguay, was lim-
ited. However, the inflata and integrifolia groups seem to
have distinctly different distributions, and populations of
the two groups were always geographically separated. Out-
side Brazil, the wetland areas between latitude 29 and 30 S
seem likely to form another geographical barrier separating
the two groups, although further research is necessary
to better define the patterns of geographical distribution.
Petunia interior T. Ando & Hashim. is a species occur-
ring on the uplands in interior SC and north-western RS
close to SC (see fig. 4 in Ando and Hashimoto, 1996).
Therefore, the distribution range of the inflata group
(Fig. 7) is close to that of P. interior in the north-western
RS. The inflata group seems to have the ability to survive as
a weed. Several dozen hectares of harvested wheat fields,
totally covered with purple flowers of the inflata group, have
often been encountered there. However, P. interior has
never been seen as a weed. It seems to adhere to undisturbed
land. In the eastern upland regions of SC and RS, another
species, P. altiplana, occurs (see fig. 5 in Ando and
Hashimoto, 1993). Petunia altiplana seems to prefer
much higher lands compared with the inflata group.
There is no Petunia species whose distribution range is
close to that of the integrifolia group except the inflata
group. Petunia axillaris whose distribution range overlaps
with those of the integrifolia and inflata groups has been
excluded from the discussion thus far. This species has large
white flowers and is considered to be reproductively isolated
from the species with purple flowers like the P. integrifolia
complex by an exclusive pollinator (Ando et al., 2001).
Comparison with the work of Wijsman
Wijsman (1982) described Petunia inflata and
P. occidentalis as subspecies of P. integrifolia. He focused
on the inflexus/deflexus position of the pedicels, a character
observed in herbarium specimens. He also emphasized the
inter-gradation of two other characters over a geographical
course from east to west, where the deflexus character chan-
ged to inflexus. The classification system used in that study,
based on the measurement of 22 herbarium specimens
belonging to the P. integrifolia complex was as follows:
(1) deflexed, (2) deflexed, but some aberrant, (3) inflexed,
but some aberrant, and (4) inflexed (see table 2 in Wijsman,
1982). However, it is believed that this classification sys-
tem is inappropriate, due to the high variability of pedicel
condition among members of the integrifolia group, as
shown in the present study (Fig. 4A and B).
Of the three specimens Wijsman (1982) classified as
(2) ‘deflexed, but some aberrant’, it was found that two
belonged to neither the integrifolia nor inflata groups,
but were in fact Petunia altiplana and a species of
Calibrachoa. The third specimen in this category (Reitz
s. n. in U) was collected from São Joaquim, SC, but it
was not possible to locate the specimen. Special attention
has been given to this region of very rich vegetation and it
has been visited 11 times (annually from 1988 to 2000,
except 1994 and 1998; Ando, 2003, 2004). From the present
observations, it can safely be concluded that Petunia
altiplana is the sole species of the genus occurring in that
region.
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In Wijsman’s (1982) map showing the geographical vari-
ation in pedicel condition for the P. integrifolia complex,
three symbols representing inflexus condition (symbol, +)
were marked in the Entre Rı́os Province of Argentina, west
of Uruguay (see fig. 2 in Wijsman, 1982). However, this is
likely to have been a mistake, since no specimens were
reported from this province (see table 2 in Wijsman,
1982). When the three symbols representing ‘deflexed,
but some aberrant’ [symbol, (–)] and the three symbols
in Entre Rı́os are removed from consideration, his map
also implies that the deflexus and inflexus conditions are
distributed in southern and northern regions, respectively.
Influenced by the instability of pedicel condition characters,
Wijsman (1982) is likely to have drawn a conclusion that
counters the true features observed in the native habitat.
Sub-cluster of the integrifolia group
The morphology of the integrifolia group seems more
variable in comparison with the inflata group, as suggested
by the larger amalgamation distances in the cluster analysis
(Fig. 2). With respect to the morphological inter-gradation
between the inflata and integrifolia groups, the existence of
a minor sub-cluster within the integrifolia group should not
be ignored, because members of the minor sub-cluster occur
mostly along the border between territories of the inflata and
integrifolia groups (closed circles in Fig. 7). In the dendro-
gram obtained from the cluster analysis (Fig. 2), a minor
sub-cluster appears to be a bridge between the cluster of the
inflata group and the major sub-cluster of the integrifolia
group. However, this is not the case. The minor sub-cluster
cannot be considered a morphological intermediate between
the inflata group and the members of the major sub-cluster.
Many of the floral organs of the minor sub-cluster members
were larger than those of the major sub-cluster (Table 3)
and distinct from those of the inflata group. As a possible
explanation for the larger floral organs, hybrid effects can-
not be discounted. Nevertheless, a hybrid swarm (B812)
was situated in the intermediate region between the inflata
and integrifolia groups (a square in Fig. 1), but the mem-
bers of the minor sub-cluster appeared as outliers of the
integrifolia group in the scatter diagram of the PC analysis
(closed circles in Fig. 1). Analysis of the minor sub-cluster
of the integrifolia group is left for future studies.
Conclusions
Wijsman (1982) emphasized an ‘apparent primary inter-
gradation’ between Petunia integrifolia and P. inflata, and
the involvement of a cline that ‘in going west, the deflexus
 Ando et al. — Petunia integrifolia Complex 897
T A B L E 4. Representative morphological traits differentiated P. inflata are differentiated in a considerable number of
between Petunia integrifolia and P. inflata morphological traits as represented in Table 4.

Morphological traits P. integrifolia P. inflata

Pedicel in the fruiting Mostly deflexed Inflexed ACKNOWLEDGEMENTS

state
Calyx lobes in the Patent and Closed and straight The authors thank Masao Udagawa of Montevideo,
fruiting state out-curved Uruguay; Sebastião T. Nagase, Nobuyuki Hiranaka,
Colour of inner Deeper than Paler than corolla-limb Tomio Koshizawa, Hideo Okubo and Roberto H. Okubo
corolla-tube corolla-limb
Darker reticulation Obscure, Restricted to upper side
of São Paulo, Brazil; and Tsuguyoshi Aoki of Buenos
Aires, Argentina, for surveying the Petunia habitats.

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on inside corolla-tube expanded widely
Long stamens Long, >16·44 mm Short, <16·42 mm
Diameter of corolla-tube Large, >9·0 mm Small, <8·5 mm
Basal part of filament Long, >5·78 mm Short, <5·23 mm
affixed to corolla-tube (95 % range) (95 % range)
approx. 3 times of approx. 2 times
ovary length of ovary length
Length along vertical Long, >1·39 mm Short, <1·12 mm
axis of stigma (95 % range) (95 % range)
character changes into inflexus’. However, no evidence of
such an inter-gradation or cline between the integrifolia
and inflata groups was found. Even omitting the pedicel
condition, a clear gap existed in the set of morphological
characteristics defining the two groups (Figs 1 and 2).
Wijsman (1982) also noted that ‘Separation of P. inflata
and P. integrifolia is very difficult’; however, based on the
characters observed here, the authors believe that it is rather
simple to distinguish the inflata and integrifolia groups
in the field. Wijsman (1982) regarded P. integrifolia and
P. inflata as ‘varieties of one biological species’, and stated,
‘The extremes can be given the following names, Petunia
integrifolia subsp. integrifolia and subsp. inflata’. The pre-
sent results disagree with these statements and suggest
that the inflata group is not merely a morphological extreme
of the integrifolia group, but rather an independent taxon
with a clearly defined morphology and distribution. Further-
more, the treatment of Petunia inflata as a synonym of
P. integrifolia (Smith and Downs, 1966) can also be safely
ruled out.
Of the three different classification systems used for
the inflata group (Fries, 1911; Smith and Downs, 1966;
Wijsman, 1982), the present results support the opinion of
Fries (1911) who referred to the inflata group as a natural
taxon (i.e. Petunia inflata) that was independent of
P. integrifolia. Of the various speciation mechanisms at
work in the plant kingdom, at least three operate in the
genus Petunia, including geographical separation (Ando
et al., 2001). The inflata and integrifolia groups are inter-
fertile (Tsukamoto et al., 1998), but interspecific genetic
exchange seems to be prevented by their different distribu-
tions (Fig. 7). The inflata group is distinguished from the
integrifolia group by at least eight site mutations in the
analysis of restriction fragment length polymorphism of
chloroplast DNA (Ando et al., 2005). Although it is neces-
sary to consult more molecular information before conclu-
sions can be drawn, the most plausible status of Petunia
inflata should be an allopatric species, or ‘geographische
Art’ (geographic species) in accordance with Fries (1911).
At this time, suffice it to say that Petunia integrifolia and
LITERATURE CITED
Ando T. 2003. Records of plant exploration for South America conducted by
the laboratory of ornamental plant science. Part I. From 1988 to 1994.
Technical Bulletin of Faculty of Horticulture, Chiba University 57:
121–135.
Ando T. 2004. Records of plant exploration for South America conducted by
the laboratory of ornamental plant science. Part II. From 1995 to 2001.
Technical Bulletin of Faculty of Horticulture, Chiba University 58:
75–91.
Ando T, Hashimoto G. 1993. Two new species of Petunia (Solanaceae)
from southern Brazil. Botanical Journal of Linnean Society 111:
265–280.
Ando T, Hashimoto G. 1994. A new Brazilian species of Petunia
(Solanaceae) from the Serra da Mantiqueira. Brittonia 46: 340–343.
Ando T, Hashimoto G. 1995. Petunia guarapuavensis (Solanaceae): a new
species from Planalto of Paraná and Santa Catarina, Brazil. Brittonia
47: 328–334.
Ando T, Hashimoto G. 1996. A new Brazilian species of Petunia
(Solanaceae) from interior Santa Catarina and Rio Grande do Sul,
Brazil. Brittonia 48: 217–223.
Ando T, Hashimoto G. 1998. Two new species of Petunia (Solanaceae)
from southern Rio Grande do Sul, Brazil. Brittonia 50: 483–492.
Ando T, Kurata M, Sasaki S, Ueda Y, Hashimoto G, Marchesi E. 1995.
Comparative morphological studies on infraspecific taxa of Petunia
integrifolia (Hook.) Schinz et Thell. (Solanaceae). Journal of Japanese
Botany 70: 205–217.
Ando T, Nomura M, Tsukahara J, Watanabe H, Kokubun H,
Tsukamoto T, et al. 2001. Reproductive isolation in a native
population of Petunia sensu Jussieu (Solanaceae). Annals of Botany
88: 403–413.
Ando T, Kokubun H, Watanabe H, Tanaka N, Yukawa T, Hashimoto G,
et al. 2005. Phylogenetic analysis of Petunia sensu Jussieu
(Solanaceae) using chloroplast DNA RFLP. Annals of Botany 96:
289–297.
Dowd PE, Mccubbin, AG, Wang X, Verica JA, Tsukamoto T, Ando T,
Kao TH. 2000. Use of Petunia inflata as a model for the study of
solanaceous type self-incompatibility. Annals of Botany (Suppl. A):
87–93.
Fries RE. 1911. Die Arten der Gattung Petunia. Kungliga Svenska
Vetenskapsakademiens Handlingar 46: 1–72.
Holmgren PK, Holmgren NH, Barnett LC. 1990. Index herbariorum.
Part I. The herbaria of the world. Bronx, NY: New York Botanical
Garden.
Hooker WJ. 1831. Salpiglossis integrifolia. Entire-leaved Salpiglossis.
Curtis’s Botanical Magazine 58: t. 3113.
Paxton J. 1836. Petunia nyctaginiflora violacea. Paxton’s Magazine of
Botany 2: 173.
Sink KC. 1984. Taxonomy. In: Sink KC, ed. Petunia. New York: Springer,
3–9.
Smith LB, Downs RJ. 1966. Petunia. In: Reitz PR, ed. Flora Illustrada
Catarinense. Solanaceas, Santa Catarina, Brazil: Herbário ‘Barbosa
Rodrigues’, Itajai, 261–291.
Tsukamoto T, Ando T, Kurata M, Watanabe H, Kokubun H,
Hashimoto G, et al. 1998. Resurrection of Petunia occidentalis
R. E. Fr. (Solanaceae) inferred from a cross compatibility study.
Journal of Japanese Botany 73: 15–21.
898 Ando et al. — Petunia integrifolia Complex
Wijsman HJW. 1982. On the inter-relationship of certain species of
Petunia. I. Taxonomic notes on the parental species of Petunia
hybrida. Acta Botanica Neerlandica 31: 477–490.
APPENDIX
Voucher specimens of the materials used in the present study
Abbreviations of the herbaria followed Holmgren et al.
(1990), except GHSP (Goro Hashimoto, Centro de Pesquisas
de História Natural, São Paulo, Brazil) and Ando (temporary
collection of Toshio Ando). An application has been made
to the New York Botanical Garden to register GHSP in the
herbaria database. It may appear in the next edition of the
Index Herbariorum.
Petunia integrifolia (Hook.) Schinz et Thell.
Argentina. Entre Rı́os: Dept Colón: Rt. N14, Arroyo
Urquiza 32 200 2100 S 58 150 4400 W, 28 Nov. 1998, T. Ando &
J. Tsukahara A168 (BAB, Ando); Dept Colón: Rt. N14,
Arroyo Concepción 31 440 2700 S 58 180 1200 W, 28 Nov.
1998, T. Ando & J. Tsukahara A171 (BAB, Ando); Dept
Concordia: Colonia Ayuı́, beside Embalse Salto Grande
31 120 0200 S 57 580 4300 W, 29 Nov. 1998, T. Ando &
J. Tsukahara A175 (BAB, Ando); Dept Gualeguaychú:
Rt. N136, Rı́o Uruguay 33 040 0400 S 58 150 2000 W, 27 Nov.
1998, T. Ando & J. Tsukahara A163 (BAB, Ando); Dept
Gualeguaychú: Gualeguaychú, beside Rı́o Gualeguaychú
33 000 2700 S 58 290 1100 W, 27 Nov. 1998, T. Ando &
J. Tsukahara A166 (BAB, Ando).
Brazil. Rio Grande do Sul: Mun. Alegrete: Rt. RS566,
Rio Ibicuı́ 29 180 3600 S 56 020 5200 W, 27 Nov. 1996,
G. Hashimoto et al. B1198 (GHSP, Ando); Mun. Boqueirão
do Leão: Rt. RS422, 4 km north-west of Boqueirão Leão
to Barros Cassal 29 160 5300 S 52 260 0900 W, 26 Nov. 1994,
G. Hashimoto et al. B870 (GHSP, Ando); Mun. Caçapava
do Sul: Rt. BR290, 3·9 km north-east-east of the junc-
tion BR153/BR290 to Pântano Grande 30 210 0300 S
53 190 5100 W, 28 Nov. 1991, G. Hashimoto et al.
B314 (BM, MBM, GHSP, Ando); Mun. Cacequi: Rt.
RS640, Cacequi 29 510 5900 S 54 490 3300 W, 7 Dec. 1993,
G. Hashimoto et al. B680 (GHSP, Ando); Mun. Cachoeira
do Sul: Rt. BR153, 22·1 km north of the junction of route
BR290 and BR153 30 040 3700 S 52 520 3200 W, 28 Nov. 1991,
G. Hashimoto et al. B315 (GHSP, Ando); Mun. Cachoeira
do Sul: Rt. BR290, 30 km west of the entrance to Cachoeira
do Sul to Vila Nova 30 170 1500 S 53 080 3200 W, 3 Dec. 1993,
G. Hashimoto et al. B651 (GHSP, Ando); Mun. Dom Pedrito:
Rt. BR293, 12 km south-east-east of Rio Ibicuı́ da Armada
to Dom Pedrito 30 520 0600 S 54 550 0300 W, 16 Nov. 1994,
G. Hashimoto et al. B802 (BM, MBM, MVFA, US,
GHSP, Ando); Mun. Encantado: Rt. RS332, Encantado
29 130 1900 S 51 530 3300 W, 27 Nov. 1995, G. Hashimoto
et al. B971 (GHSP, Ando); Mun. Lavras do Sul: 29 km
north-east of Dom Pedrito to Ibaré 30 510 0100 S
54 280 1900 W, 25 Nov. 1996, G. Hashimoto et al. B1190
(GHSP, Ando); Mun. Mata: 3 km south-west of Mata
to BR287 29 330 5300 S 54 280 1600 W, 21 Nov. 1994,
G. Hashimoto et al. B835 (GHSP, Ando); Mun. Pântano
Grande: Rt. BR471, 10 km north of Pântano Grande to
Rio Pardo 30 100 0600 S 52 220 2500 W, 25 Nov. 1994,
G. Hashimoto et al. B867 (GHSP, Ando); Mun. Quaraı́:
Rt. BR293, 1·6 km north-west of Rio Cati to Quaraı́
30 290 0900 S 56 130 0600 W, 27 Nov. 1991, G. Hashimoto
et al. B300 (GHSP, Ando); Mun. Quaraı́: Rt. BR293,
28·0 km south-east of Quaraı́ to Santana do Livramento
30 270 4600 S 56 160 1100 W, 27 Nov. 1991, G. Hashimoto
et al. B301 (GHSP, Ando); Mun. Restinga Seca: Rt.
RS509, 12 km west of Rio Jacuı́ to Santa Maria
29 430 2600 S 53 240 0300 W, 26 Nov. 1995, G. Hashimoto
et al. B948 (GHSP, Ando); Mun. Rio Pardo: Rt. BR471,
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3 km south of Rio Pardo to Pântano Grande 30 000 2800 S
52 210 5900 W, 25 Nov. 1994, G. Hashimoto et al. B868
(GHSP, Ando); Mun. Rosário do Sul: Rt. BR158, 10 km
south-west of the junction BR158/BR290 to Santana
do Livramento 30 200 0600 S 55 000 3500 W, 6 Dec. 1993,
G. Hashimoto et al. B673 (GHSP, Ando); Mun. Rosário
do Sul: Rt. RS640, 15 km north of the junction BR290/
RS640 to Cacequi 30 070 1800 S 54 490 2800 W, 7 Dec. 1993,
G. Hashimoto et al. B677 (GHSP, Ando); Mun. Rosário
do Sul: 20 km north-west-west of Virador to São Leandro
30 220 4300 S 55 250 1900 W, 22 Nov. 1994, G. Hashimoto et al.
B841 (GHSP, Ando); Mun. Santa Maria: Rt. BR392, 21·6 km
south-east of the entrance of Santa Maria 29 530 2500 S
53 430 5300 W, 25 Nov. 1991, G. Hashimoto et al. B280
(GHSP, Ando); Mun. Santa Maria: Rt. BR287, 8 km east
of Rio Ibicuı́ to Santa Maria 29 400 1400 S 54 010 2800 W,
18 Nov. 1994, G. Hashimoto et al. B818 (GHSP, Ando);
Mun. Santana do Livramento: Rt. BR293, 15·5 km west
of the junction BR293/BR158 30 490 3300 S 55 200 1500 W,
27 Nov. 1991, G. Hashimoto et al. B294 (MVFA, GHSP,
Ando); Mun. Santiago: Rt. BR287, 22 km north-north-west
of Jaguari to Santiago 29 210 0500 S 54 450 2900 W, 7 Dec. 1993,
G. Hashimoto et al. B686 (GHSP, Ando); Mun. Santiago: Rt.
BR287, 18 km south-east of Santiago to Jaguari 29 160 3000 S
54 470 4200 W, 7 Dec. 1993, G. Hashimoto et al. B687 (GHSP,
Ando); Mun. Santiago: Rt. BR287, 2 km north of the south
entrance of Santiago 29 110 4900 S 54 500 5600 W, 7 Dec. 1993,
G. Hashimoto et al. B688 (GHSP, Ando); Mun. Santiago:
Rt. BR287, 30 km south-east of Encruzilhada to Santiago
29 020 2600 S 55 140 4900 W, 8 Dec. 1993, G. Hashimoto et al.
B705 (GHSP, Ando); Mun. Santiago: Rt. BR287, 42 km
south-east of Encruzilhada to Santiago 29 030 0500 S
55 070 3600 W, 8 Dec. 1993, G. Hashimoto et al. B707
(GHSP, Ando); Mun. Santiago: Rt. BR287, 9 km north-
west of the junction BR287/road to São Luı́z Gonzaga
29 030 2900 S 54 570 4200 W, 8 Dec. 1993, G. Hashimoto et al.
B708 (GHSP, Ando); Mun. Santiago: Rt. RS546, Santiago
29 120 1400 S 54 530 0200 W, 9 Nov. 1997, G. Hashimoto
et al. B1304 (GHSP, Ando); Mun. São Borja: Rt. BR287,
7 km south-east of Nhuporã to Encruzilhada 28 490 5300 S
55 430 4200 W, 8 Dec. 1993, G. Hashimoto et al. B701
(GHSP, Ando); Mun. São Borja: Rt. BR287, 19 km south-
east of Nhuporã to Encruzilhada 28 540 1600 S 55 380 1600 W,
8 Dec. 1993, G. Hashimoto et al. B702 (GHSP, Ando);
Mun. São Borja: Rt. BR287, 11 km east of Encruzilhada
to Santiago 28 580 4600 S 55 230 4500 W, 8 Dec. 1993,
G. Hashimoto et al. B704 (GHSP, Ando); Mun.
São Francisco de Assis: Rt. RS241, 8 km north-west of
Rio Jaguarı́ to São Francisco de Assis 29 380 2600 S
 Ando et al. — Petunia integrifolia Complex
55 000 5900 W, 26 Nov. 1996, G. Hashimoto et al. B1192
(GHSP, Ando); Mun. São Francisco de Assis: Rt. RS241,
8 km south-east of São Francisco de Assis to Rio
Jaguarı́ 29 360 1500 S 55 040 3500 W, 26 Nov. 1996,
G. Hashimoto et al. B1193 (GHSP, Ando); Mun. São
Francisco de Assis: Rt. RS241, 16 km east of Manuel
Viana to São Francisco de Assis 29 340 2000 S 55 200 4000 W,
26 Nov. 1996, G. Hashimoto et al. B1195 (GHSP, Ando);
Mun. São Francisco de Assis: Rt. RS176, 31 km north of
Manuel Viana to Rio Itú 29 190 1300 S 55 260 4900 W, 9 Nov.
1997, G. Hashimoto et al. B1314 (GHSP, Ando); Mun.
São Gabriel: Rt. BR290, 28·8 km north-west of entrance
of São Gabriel to Rosário do Sul 30 150 0000 S 54 340 5900 W,
28 Nov. 1991, G. Hashimoto et al. B311 (GHSP, Ando);
Mun. São Gabriel: Rt. BR290, 9 km east of São Gabriel
to Vila Nova 30 210 5200 S 54 140 2900 W, 23 Nov. 1994,
G. Hashimoto et al. B848 (GHSP, Ando); Mun. São Pedro
do Sul: Rt. BR287, 7 km west of entrance of São Pedro do
Sul to São Vicente do Sul 29 380 0300 S 54 150 4200 W,
18 Nov. 1994, G. Hashimoto et al. B817 (GHSP, Ando);
Mun. São Vicente do Sul: Rt. BR287, 5 km north of
São Vicente do Sul to Jaguari 29 380 1900 S 54 400 4500 W,
7 Dec. 1993, G. Hashimoto et al. B682 (GHSP, Ando);
Mun. Sobradinho: 15 km south-east of Sobradinho
to Candelária 29 300 0600 S 52 550 4500 W, 26 Nov. 1995,
G. Hashimoto et al. B949 (GHSP, Ando); Mun. Uruguaiana:
Rt. BR472, 3 km south-west of João Arrengui to Uruguaiana
29 290 5500 S 56 410 4700 W, 17 Nov. 1994, G. Hashimoto et al.
B809 (GHSP, Ando).
Uruguay. Flores: Rt. 3, 1·9 km north-west of Andresito
33 090 1900 S 57 080 3100 W, 13 Nov. 1990, T. Ando & K. Buto
U168 (MVFA, SI, GHSP, Ando).
Rı́o Negro: Rt. 2, 1·6 km north-west of the bridge over Rı́o
Negro to Fray Bentos 33 130 2800 S 58 010 3100 W, 28 Nov. 1989,
T. Ando & H. Watanabe U106 (MVFA, S, SI, GHSP, Ando);
Parque Puerto Viejo, N of San Javier 32 300 1300 S
58 080 5000 W, 12 Nov. 1990, T. Ando & K. Buto U161
(BM, MVFA, S, SI, US, GHSP, Ando); Rt. 3, 1 km north-
west of the bridge over Rı́o Negro 33 070 4600 S 57 100 1900 W,
4 Nov. 1991, T. Ando & S. Iida U239 (MVFA, Ando);
Las Canas 33 100 1300 S 58 210 1800 W, 14 Dec. 1992,
T. Ando & K. Shibata U263 (BM, MVFA, S, US, Ando).
Rivera: Rt. 5, 7·2 km south of junction Rt. 5/30 to
Tacuarembó 31 180 2700 S 55 400 2200 W, 18 Nov. 1988, T.
Ando & H. Kokubun U12 (BM, MVFA, Ando).
Salto: Rt. 3, entrance of Termas de Arapey 30 540 2700 S
57 41 3700 W, 3 Nov. 1991, T. Ando & S. Iida U231 (MVFA,
 0
Ando); 5 km south-west of Salto 31 230 4900 S 57 590 0700 W,
4 Nov. 1991, T. Ando & S. Iida U233 (BM, MVFA, Ando).
Tacuarembó: San Gregorio de Polanco 32 370 2900 S
55 49 5400 W, 23 Nov. 1988, T. Ando & H. Kokubun
 0
U46 (MVFA, Ando); Gruta de los Cuelvos, north-west of
Tacuarembó, 30 Nov. 1989, T. Ando & H. Watanabe U128
(MVFA, Ando).
Petunia inflata R. E. Fr.
Argentina. Chaco: Dept Quitilipi: Rt. P4, 7·4 km south-
south-west of Pampa Verde to Quitilipi 26 310 1600 S
60 040 2900 W, 20 Nov. 1999, T. Ando et al. A260 (BAB,
Ando).
899
Corrientes: Dept Empedrado: Rt. N12, 8·2 km north-north-
west of Arroyo San Lorenzo to Empedrado 28 020 4300 S
58 470 5700 W, 21 Nov. 1999, T. Ando et al. A270 (BAB,
Ando).
Formosa: Dept Pirané: Rt. N81, 11·0 km north-west of
entrance of Pirané to Palo Santo 25 380 0700 S 59 090 0400 W,
17 Nov. 1999, T. Ando et al. A249 (BAB, Ando).
Misiones: Dept Candelaria: Rt. N4, Parque Central,
Bonpland 27 290 0500 S 55 280 4100 W, 27 Nov. 1990, T.
Ando & K. Buto A7 (SI, GHSP, Ando); Dept San Ignacio:
Rt. RN12, 6·2 km north-east of Santo Pipó to Jardin América
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27 060 5100 S 55 210 3200 W, 10 Nov. 1995, T. Ando &
T. Tsukamoto A119 (BAB, Ando).
Brazil. Rio Grande do Sul: Mun. Arvorezinha:
10 km west of Morangueira to Soledade 28 410 5400 S
52 140 4600 W, 21 Nov. 1996, G. Hashimoto et al. B1163
(GHSP, Ando); Mun. Barros Cassal: 17 km south-west
of Barros Cassal to Segredo 29 080 3300 S 52 420 3800 W,
26 Nov. 1995, G. Hashimoto et al. B956 (GHSP, Ando);
Mun. Casca: Rt. RS324, 11 km west of Casca to Marau
28 330 0400 S 52 040 1400 W, 28 Nov. 1993, G. Hashimoto
et al. B587 (GHSP, Ando); Mun. Cirı́aco: Rt. BR285,
27 km west of Lagoa Vermelha to Cruz Altinha
28 140 5700 S 51 440 3200 W, 28 Nov. 1993, G. Hashimoto
et al. B581 (GHSP, Ando); Mun. Cruz Alta: Rt. RS342,
Cruz Alta 28 380 3700 S 53 370 5100 W, 25 Nov. 1993,
G. Hashimoto et al. B540 (GHSP, Ando); Mun. Erexim:
Rt. BR153, 1 km south of the junction BR153/RS331
27 370 3000 S 52 140 0700 W, 1 Dec. 1996, G. Hashimoto et al.
B1222 (GHSP, Ando); Mun. Getúrio Vargas: Rt. RS135,
9·7 km north of north entrance of Getúlio Vargas to Erexim
27 490 2100 S 52 170 3200 W, 23 Nov. 1991, G. Hashimoto et al.
B261 (GHSP, Ando); Mun. Giruá: Rt. RS544, 13 km
south-south-west of Santa Rosa to Guarani das Missões
27 580 4900 S 54 320 3500 W, 29 Nov. 1996, G. Hashimoto
et al. B1211 (GHSP, Ando); Mun. Guarani das Missões:
Guarani das Missões 28 090 0100 S 54 330 0200 W, 29 Nov.
1996, G. Hashimoto et al. B1209 (GHSP, Ando); Mun.
Guarani das Missões: Rt. RS544, 3 km north of Guarani
das Missões to Santa Rosa 28 070 4300 S 54 330 4300 W,
29 Nov. 1996, G. Hashimoto et al. B1210 (GHSP, Ando);
Mun. Ibiaçá: Rt. RS126, entrance to Ibiaçá 28 010 3700 S
51 460 3300 W, 9 Dec. 1993, G. Hashimoto et al. B724
(GHSP, Ando); Mun. Ijuı́: Rt. BR285, 6·7 km west of
Rio Conceição to Santo Ângelo 28 220 4000 S 54 050 4600 W,
24 Nov. 1991, G. Hashimoto et al. B272 (GHSP, Ando);
Mun. Júlio de Castilhos: 39 km south-east-east of Júlio de
Castilhos to Pinhal Grande 29 210 0600 S 53 240 3000 W, 25 Nov.
1993, G. Hashimoto et al. B544 (MBM, GHSP, Ando);
Mun. Júlio de Castilhos: 25 km north-east of Pinhal Grande
to Barragem de Itaúba 29 150 3400 S 53 140 5200 W, 25 Nov.
1993, G. Hashimoto et al. B549 (GHSP, Ando); Mun.
Machadinho: 2 km north of Machadinho to Capinzal (SC)
27 320 5500 S 51 390 5200 W, 27 Nov. 1993, G. Hashimoto et al.
B574 (GHSP, Ando); Mun. Nonoai: Rt. RS406, 14 km south-
west of Nonoai to Trinidade 27 260 3600 S 52 530 0600 W,
26 Nov. 1993, G. Hashimoto et al. B564 (GHSP, Ando);
Mun. Palmeira das Missões: Rt. BR468, 15 km north-west
of Palmeira das Missões to Coronel Bicaco 27 490 1400 S
900 Ando et al. — Petunia integrifolia Complex
53 250 0200 W, 24 Nov. 1993, G. Hashimoto et al. B522
(GHSP, Ando); Mun. Palmeira das Missões: Rt. RS569,
8 km east of Palmeira das Missões to Sarandi 27 530 3700 S
53 140 2000 W, 7 Nov. 1997, G. Hashimoto et al. B1270
(GHSP, Ando); Mun. Panambi: Rt. BR285, 6·1 km west of
the junction BR285/RS158 to Ijuı́ 28 210 4300 S 53 360 5800 W,
24 Nov. 1991, G. Hashimoto et al. B269 (MVFA, GHSP,
Ando); Mun. Santa Rosa: Rt. RS544, 4 km south-west
of Santa Rosa to Guarani das Missões 27 540 4400 S
54 300 2900 W, 29 Nov. 1996, G. Hashimoto et al. B1212
(GHSP, Ando); Mun. Santo Ângelo: São Miguel
das Missões 28 320 5500 S 54 330 3600 W, 24 Nov. 1991,
G. Hashimoto et al. B274 (GHSP, Ando); Mun. Santo
Ângelo: Rt. RS344, 14 km north-north-west of Santo
Ângelo to Giruá 28 110 4500 S 54 190 2100 W, 24 Nov. 1993,
G. Hashimoto et al. B538 (GHSP, Ando); Mun. Santo
Antônio das Missões: Rt. BR285, 6 km west of the entrance
of Santo Antônio das Missões to Itaroquém 28 290 3300 S
55 160 5300 W, 8 Dec. 1993, G. Hashimoto et al. B692
(GHSP, Ando); Mun. Santo Antônio das Missões: Rt.
BR285, 7 km west of Santo Antônio das Missões to São
Borja 28 290 3600 S 55 170 4200 W, 8 Dec. 1993, G. Hashimoto
et al. B694 (GHSP, Ando); Mun. Santo Antônio das Missões:
Rt. BR285, 19 km NEE of Rio Icamaquã to Itaroquém
28 330 0300 S 55 340 0000 W, 8 Dec. 1993, G. Hashimoto et al.
B697 (GHSP, Ando); Mun. Santo Antônio das Missões: Rt.
BR285, 10 km north-east-east of Rio Icamaquã to Itaroquém
28 350 3600 S 55 380 4100 W, 8 Dec. 1993, G. Hashimoto et al.
B698 (GHSP, Ando); Mun. Santo Antônio das Missões: Rio
Piratinim, between São Nicolau and Garruchos 28 120 2900 S
55 190 1000 W, 28 Nov. 1996, G. Hashimoto et al. B1205
(GHSP, Ando); Mun. Sarandi: Rt. RS404, 7 km north of
Sarandi to Rondinha 27 530 2000 S 52 540 5000 W, 26 Nov.
1993, G. Hashimoto et al. B560 (GHSP, Ando); Mun. São
Borja: 8 km north-east of Rincão do Meio to Garruchos
28 260 4600 S 55 310 2200 W, 28 Nov. 1996, G. Hashimoto
et al. B1200 (GHSP, Ando); Mun. São Borja: 24 km
south-east of Garruchos to Rincão do Meio 28 170 5300 S
55 310 4900 W, 28 Nov. 1996, G. Hashimoto et al. B1202
(GHSP, Ando); Mun. São Borja: Garruchos 28 100 5600 S
55 380 3100 W, 28 Nov. 1996, G. Hashimoto et al. B1203
(GHSP, Ando); Mun. São Borja: 24 km south-east of
Garruchos to Santo Antônio das Missões 28 150 1800 S
55 260 2500 W, 28 Nov. 1996, G. Hashimoto et al. B1204
(GHSP, Ando); Mun. São Luı́z Gonzaga: Rt. BR285, 3 km
east of São Luı́z Gonzaga to Santo Ângelo 28 250 0100 S
54 550 5300 W, 8 Dec. 1993, G. Hashimoto et al. B709
(GHSP, Ando); Mun. São Luı́z Gonzaga: Rt. BR285,
28 km E of São Luı́z Gonzaga to Santo Ângelo
28 240 1400 S 54 400 5400 W, 8 Dec. 1993, G. Hashimoto et al.
B710 (GHSP, Ando); Mun. São Luı́z Gonzaga: Rt.
RS544, 3 km north-east of Rolador to Cerro Largo
28 140 3400 S 54 470 3600 W, 29 Nov. 1996, G. Hashimoto
et al. B1208 (GHSP, Ando); Mun. São Nicolau: 8 km
south-east of São Nicolau to São Luı́z Gonzaga
28 130 0700 S 55 120 1900 W, 28 Nov. 1996, G. Hashimoto
et al. B1206 (GHSP, Ando); Mun. São Valentim: Rt.
RS480, 5 km north-west of São Valentim to Erval Grande
27 310 4500 S 52 320 2000 W, 1 Dec. 1996, G. Hashimoto et al.
B1219 (GHSP, Ando); Mun. Segredo: Lagoão 29 140 0900 S
52 470 4600 W, 26 Nov. 1995, G. Hashimoto et al. B954
(GHSP, Ando); Mun. Selbach: Rt. RS153, 12 km east of
Ibirubá to Tapera 28 380 3900 S 52 580 1000 W, 9 Dec. 1993,
G. Hashimoto et al. B719 (GHSP, Ando); Mun. Tapejara:
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8 km north-east of Tapejara to Ibiaçá 28 010 2800 S
51 570 2700 W, 9 Dec. 1993, G. Hashimoto et al. B723
(GHSP, Ando); Mun. Três de Maio: Rt. RS210, 11 km
west of Três de Maio to Santa Rosa 27 460 2300 S
54 200 0500 W, 24 Nov. 1993, G. Hashimoto et al. B531
(GHSP, Ando); Mun. Tupanciretã: Rt. BR158, 10 km
north of entrance of Tupanciretã to Cruz Alta 28 560 4400 S
53 380 4900 W, 7 Nov. 1997, G. Hashimoto et al. B1287
(GHSP, Ando); Mun. Tupanciretã: 8 km east of Tupanciretã
to Rt. BR158 29 030 1600 S 53 460 0100 W, 8 Nov. 1997,
G. Hashimoto et al. B1288 (GHSP, Ando); Mun.
Tupanciretã: Rt. BR377, 78 km west of Tupanciretã to
Santiago 29 010 0100 S 54 230 3500 W, 8 Nov. 1997,
G. Hashimoto et al. B1302 (GHSP, Ando); Mun. Tuparendi:
Rt. RS344, 4 km north-west of Tuparendi to Porto Mauá
27 440 3200 S 54 300 3100 W, 24 Nov. 1993, G. Hashimoto
et al. B533 (GHSP, Ando); Mun. Victor Graeff: Rt.
BR386, 12 km north-north-west of Rio Ijuı́ to Carazinho
28 300 3500 S 52 380 2200 W, 26 Nov. 1993, G. Hashimoto
et al. B556 (GHSP, Ando).
Santa Catarina: Mun. Anita Garibaldi: Rt. SC458, 16 km
north-west of Cerro Negro to Anita Garibaldi 27 450 1600 S
51 000 2500 W, 30 Nov. 1993, G. Hashimoto et al. B628
(GHSP, Ando); Mun. Campos Novos: Rt. SC456,
5·8 km north of junction BR470/SC456 to Monte Carlo
27 170 3100 S 50 580 3300 W, 3 Dec. 1991, G. Hashimoto et al.
B350 (GHSP, Ando); Mun. Itapiranga: Itapiranga
27 100 5200 S 53 430 1000 W, 25 Nov. 1992, G. Hashimoto
et al. B377 (GHSP, Ando); Mun. São José do Cerrito: Rt.
BR282, 8 km south-east of Rio Canoas to São José do Cerrito
27 340 2900 S 50 480 1500 W, 29 Nov. 1992, G. Hashimoto et al.
B439 (GHSP, Ando).
Paraguay. Central: Rt. 1, Itá, 28 Jul 1990, T. Ikemizu &
T. Ando P1 (Ando).
Intermediate form between P. integrifolia and
P. inflata.
Brazil. Rio Grande do Sul: Mun. São Borja: 10 km north of
Capitão Porto Alegre to São José 28 510 2900 S 55 320 1200 W,
18 Nov. 1994, G. Hashimoto et al. B812 (GHSP, Ando).