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A Morphological Study of The Petunia Integrifolia Complex (Solanaceae)
A Morphological Study of The Petunia Integrifolia Complex (Solanaceae)
A Morphological Study of The Petunia Integrifolia Complex (Solanaceae)
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.
PC analysis
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
†
After Ando et al. (1995).
z
Coefficient of variation (%).
x
Level of significance by t-test at: ***, 0·1 %; **, 1·0 %.
A X6 B X6
X16 X16
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
†
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).
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
Pedicel condition
Frequency
10
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- 0
40 65 90 115 140 165 190 215
ded from herbarium specimens showed a narrower range
(48–150 ; mean 108 ) compared with cultivated plants. The Angle of pedicel to stem during fruiting (degrees)
typical features of the pedicel characters are shown in in herbarium specimen
Fig. 5A (integrifolia group) and B (inflata group). F I G . 4. Frequency distributions of mean pedicel angle: (A) measured from
cultivated live plants; (B) measured from herbarium specimens.
Another character observable in herbarium specimens
The calyx lobes located next to the maturing capsules of
specimens belonging to the integrifolia group were patent group was found in upland areas in northern RS and Santa
and out-curved, whereas those of the inflata group were Catarina (SC) states, whereas the integrifolia group was
closed and straight. This subtle difference changed the found in lowland areas, such as along the Rio Ibicuı́ flow-
appearance of the calyx lobes, and was easily observed ing west, and the Rio Jacuı́ and its upper stream (the Rio
in the herbarium specimens (Fig. 5A and B). In most Vacacaı́), which both flow east (Fig. 7).
cases, at least one of the five calyx lobes of the integrifolia Outside Brazil, the inflata group was the sole taxa found
group was folded (see arrows, Fig. 5A). By contrast, all the in the Chaco, Corrientes, Formosa and Misiones Provinces
calyx lobes of the inflata group were usually straight of Argentina, and Paraguay while the integrifolia group was
(Fig. 5B). The frequency distribution of the mean values found exclusively in the Entre Rı́os Province of Argentina,
is shown in Fig. 6. and Uruguay. Compared with Brazil, the range of the inflata
group extended much further north in Paraguay and Argen-
Geographic distribution tina, up to 25 S (Fig. 7). In Corrientes Province, the range of
the inflata group was restricted to the northern region, close
Figure 7 shows a topographical map (200-m intervals) of
to Paraguay. The range of the integrifolia group was con-
the study region, indicating the location of populations
centrated along major rivers, such as the Rı́o Uruguay and
of the integrifolia (open and solid circles) and inflata
Rı́o Negro (Fig. 7).
(triangles) groups, and population B812 (solid square).
The distribution range of the inflata group clearly differed
Sub-clusters of the integrifolia group
from that of the integrifolia group. The inflata group
was located in northern areas, whereas the integrifolia The dendrogram (Fig. 2) shows that amalgamation
group was located in southern areas. In Brazil, the inflata distances were smaller in the inflata group than in the
Ando et al. — Petunia integrifolia Complex 893
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
DISCUSSION
integrifolia group. In the integrifolia cluster, a minor sub-
Identification of live plants
cluster composed of five populations (B835, B841, B870,
B949 and B971) was recognized as an outlier (Fig. 2, solid Although the taxonomic treatment of the integrifolia and
circles). Compared with the major sub-cluster, members inflata groups is complicated as mentioned in the Introduc-
of the minor sub-cluster were characterized by significantly tion, it was possible to distinguish the two groups in their
larger flowers (X21 and X25), internal floral organs (X5, X6, natural habitats. The accuracy of the authors’ tentative
X8, X9 and X10) and capsules (X30 and X31). This trend to identification, based mainly on overall flower shape, colour
larger organs was also evident for some other characters, and venation patterns, was validated by the clear statistical
such as X7 and X19 (Table 3). In addition, members of the separation of two groups using PC and cluster analyses
minor sub-cluster had pedicel angles during fruiting (X34) (Figs 1 and 2), with the exception of a probable hybrid
of around 90 (Table 3). swarm (B812 population).
Although members of the major sub-cluster of the Aside from the character of pedicel condition in the
integrifolia group always occurred in lowland habitats, fruiting state, which was not included in the multivariate
894 Ando et al. — Petunia integrifolia Complex
s
ione
26°S 26°S
Mi s
PARAGUAY m
200 0m
40 0m
100
Chaco
tarina
0m
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
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- suggesting that characters X6 and X8 are stable in the inflata
ments distinguished members of the inflata and integrifolia group irrespective of the flower size, which can vary
groups. The inflata group was characterized by significantly between populations in different local environmental con-
shorter (or smaller) inner floral organs, such as stamens ditions. It is also noteworthy that two secondary characters,
(X5), basal part of the filament fixed to the corolla tube X7 [proportion of X6 to the length of long stamen (X5)] and
(X6), pistil (X8), ovary (X16), calyx lobe (X12), calyx tube X19 [ratio of X6 : the length of ovary (X16)], which are not
(X14) and stigma (X9 and X10) (Table 1). The ranges of three directly related to flower size, also differed significantly
characters (X5, X8, and X24) for the inflata group did not between the inflata and integrifolia groups (Table 1).
overlap those of the integrifolia group. The difference in character X19 showed that the basal
In the integrifolia group, the size of the inner floral organs part of the filament affixed to the corolla tube (X6) is
(X5, X6, X8 and X16) produced higher CCs with flower size 1·98 (approx. 2) and 2·93 (approx. 3) times longer than
[as assessed by the length of the corolla (X25) or corolla tube the length of the ovary (X16) in the inflata and integrifolia
(X21)]. In the inflata group, the length of the basal part groups, respectively (Table 1 and Fig. 3). From experience,
of the filament affixed to the corolla-tube (X6) and the length it is suggested that character X19 is one of the most con-
of pistil (X8) did not exhibit these features (Table 2), 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
†
Level of significance by t-test at: ***, 0·1 %; **, 1·0 %; *, 5·0 %.
integrifolia groups. Unfortunately, many of the characters Petunia integrifolia complex can be identified as belonging
that differentiate the two groups are fragile and are not to the inflata group when most of the capsules have straight
conserved in herbarium specimens. calyx lobes without folding, since this feature is rarely
observed in the integrifolia group (Fig. 6).
The identification of the P. integrifolia complex, as
Identification in the herbaria
described above, can be applied to mature plants; however,
In previous studies, Fries (1911) and Wijsman (1982) it is often much more difficult to identify younger plants and
highlighted the pedicel condition in the fruiting state as plants grown in shade. For mature plants, another index
the primary feature distinguishing the inflata and integrifo- character can be used. The upper leaves of mature plants
lia groups. By contrast, it was found that this character (X33) belonging to the inflata group tend to be very small and
is variable in the integrifolia group, as indicated by the large scale-like (see the stem on the right-hand side of Fig. 5B).
coefficient of variation (CV) value (Table 1). In fact, char- This feature does not occur in the integrifolia group
acters related to pedicel condition measured from cultivated (Fig. 5A). Based on this morphological feature, the inflata
members of the integrifolia group (Fig. 4A) differed mark- and integrifolia groups can be distinguished without requir-
edly from those measured on voucher herbarium specimens ing the inspection of inner floral organs.
(Fig. 4B). Lateral stems of both subspecies are usually
ascending under natural conditions, but in the present
Distribution
study, lateral stems were maintained in a vertical position
with the aid of plastic supports. Perhaps the manipulation of In Brazil, the integrifolia and inflata groups were isolated
the lateral stems caused this discrepancy. geographically (Fig. 7). The inflata group occurred in north-
For herbarium specimens, the pedicel angle for the inflata ern upland areas, on a lava plateau called the Planalto. This
group fell into a rather narrow range of <80 (Fig. 4B). region is characterized by rather flat, gentle hilly ground.
Therefore, it is reasonable to conclude that the inflata The steep slope at the edge of Planalto is known as the Serra
group has inflexed pedicels at the fruiting stage. The pedicel Geral. The elevation of the Planalto exceeds 1000 m at the
angle for integrifolia was more variable and did not always eastern end close to the Atlantic coast, and the Serra Geral
exceed 90 (Fig. 4B). As a result, these cannot be considered facing the east forms a perpendicular cliff. To the west, the
as reliable taxonomic features. A member of the Petunia elevation of the Planalto gradually decreases, and the Serra
integrifolia complex can be identified as belonging to the Geral facing south is strongly eroded to form a complex
integrifolia group when it has deflexed pedicels, since this terrain. The limit between the Planalto and the lowland area,
characteristic does not occur in the inflata group (Fig. 4B). i.e. the border between the territories of the inflata and
If a specimen has a slightly inflexed pedicel, however, other integrifolia groups, is evident in central RS, but becomes
characters must be considered carefully before a clear obscure in western RS. No geographical border, limiting the
identification can be made. territories of the inflata and integrifolia groups seems to
In this study, another morphological character was found exist near the town of São Borja (indicated with a star in
that is readily observable in herbarium specimens and can Fig. 7), although the territories of the two groups are close to
be used to distinguish the two groups. For the integrifolia each other at that point.
group at least one of the five calyx lobes is usually folded The region surrounding São Borja was visited four times
(Fig. 5A), whereas in the inflata group, all the calyx lobes (1993, 1994, 1996 and 1997) to study distribution (Ando,
are straight (Fig. 5B). It is worth noting that this character 2003, 2004). The smallest distance between populations of
is stable in the integrifolia group, but more variable in the the integrifolia (B701, Mun. São Borja) and inflata (B699,
inflata group (Fig. 6), contrary to pedicel condition, as Mun. São Borja, route BR285, 5 km west-south-west of Rio
mentioned above (Fig. 4). A herbarium specimen of the 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 belonged to neither the integrifolia nor inflata groups,
possible hybrid swarm, was situated within the territory of but were in fact Petunia altiplana and a species of
the integrifolia group (solid square in Fig. 7), but only 30 km Calibrachoa. The third specimen in this category (Reitz
from a population of the inflata group (B698, Mun. Santo s. n. in U) was collected from São Joaquim, SC, but it
Antônio das Missões). The Rio Icamaquã, which flows close was not possible to locate the specimen. Special attention
to São Borja, appears as another border on the map (Fig. 7), has been given to this region of very rich vegetation and it
although this river seems too small to function as an obs- has been visited 11 times (annually from 1988 to 2000,
tacle capable of physically separating the two groups. except 1994 and 1998; Ando, 2003, 2004). From the present
Most of the research in this study was conducted in observations, it can safely be concluded that Petunia
Brazil, and the number of populations assessed from other altiplana is the sole species of the genus occurring in that
inland locations, such as Argentina and Paraguay, was lim- region.