Leaf morphology and venation of Psidium species from the Brazilian Savanna

Abstract

The Brazilian Savanna biome has the largest plant diversity among savannas worldwide and is the predominant biome in Goiás state, Brazil. Among plant species previously catalogued in the Brazilian Savanna, the Myrtaceae family has received attention, as these plants show great economic potential for its medicinal properties, food and ecological importance, highlighting in this context the Psidium genus. In order to contribute to the differentiation of problematic taxonomic groups, such as Myrtaceae, and to quality control of its plant material, morphological and venation leaf studies of four species of Psidium were performed. For this purpose, leaf samples of Psidium firmum O.Berg., Psidium myrsinites DC., Psidium laruotteanum Cambes., and Psidium guineense Sw. were collected from Goiás State University, Anápolis Air Base, and Serra de Caldas Novas State Park and submitted to classical techniques for morphological and leaf venation characterizations. The results showed that P. firmum presents brochidodromous secondary veins, marginal last venation of the fimbrial type, an abaxial surface with a grooved midrib, flat secondary veins on both sides, an apex obtuse to mucronate, and a rounded base. P. guineense presents trichomes on both surfaces, a grooved midrib on the adaxial surface and a prominent midrib on the abaxial surface, which distinguishes this species from all other Myrtaceae species examined in the present study. The species P. guineense and P. firmum presented a set of differential leaf characteristics among the others taxa of the genus, clearly separating these plants in the morphological identification key.

Full text

Revista
Brasileira
de
Farmacognosia
27
(2017)
407–413
ww
w.elsevier.com/locate/bjp
Original
Article
Leaf
morphology
and
venation
of
Psidium
species
from
the
Brazilian
Savanna
Elaine
F.
Oliveiraa,
Debborah
G.
Bezerraa,
Mirley
L.
Santosa,
Maria
H.
Rezendeb,
Joelma
A.M.
Paulaa,∗
aUniversidade
Estadual
de
Goiás,
Campus
Anápolis
de
Ciências
Exatas
e
Tecnológicas,
Anápolis,
GO,
Brazil
bDepartamento
de
Botânica,
Instituto
de
Ciências
Biológicas,
Universidade
Federal
de
Goiás,
Goiânia,
GO,
Brazil
a
r
t
i
c
l
e
i
n
f
o
Article
history:
Received
1
December
2016
Accepted
6
March
2017
Available
online
14
July
2017
Keywords:
“Arac¸
á”
Diaphanization
Leaf
venation
Taxonomy
Quality
control
a
b
s
t
r
a
c
t
The
Brazilian
Savanna
biome
has
the
largest
plant
diversity
among
savannas
worldwide
and
is
the
predominant
biome
in
Goiás
state,
Brazil.
Among
plant
species
previously
catalogued
in
the
Brazilian
Savanna,
the
Myrtaceae
family
has
received
attention,
as
these
plants
show
great
economic
potential
for
its
medicinal
properties,
food
and
ecological
importance,
highlighting
in
this
context
the
Psidium
genus.
In
order
to
contribute
to
the
differentiation
of
problematic
taxonomic
groups,
such
as
Myrtaceae,
and
to
quality
control
of
its
plant
material,
morphological
and
venation
leaf
studies
of
four
species
of
Psidium
were
performed.
For
this
purpose,
leaf
samples
of
Psidium
firmum
O.Berg.,
P.
myrsinites
DC.,
P.
laruot-
teanum
Cambes.,
and
P.
guineense
Sw.
were
collected
from
Goiás
State
University,
Anápolis
Air
Base,
and
Serra
de
Caldas
Novas
State
Park
and
submitted
to
classical
techniques
for
morphological
and
leaf
vena-
tion
characterizations.
The
results
showed
that
P.
firmum
presents
brochidodromous
secondary
veins,
marginal
last
venation
of
the
fimbrial
type,
an
abaxial
surface
with
a
grooved
midrib,
flat
secondary
veins
on
both
sides,
an
apex
obtuse
to
mucronate,
and
a
rounded
base.
P.
guineense
presents
trichomes
on
both
surfaces,
a
grooved
midrib
on
the
adaxial
surface
and
a
prominent
midrib
on
the
abaxial
surface,
which
distinguishes
this
species
from
all
other
Myrtaceae
species
examined
in
the
present
study.
The
species
P.
guineense
and
P.
firmum
presented
a
set
of
differential
leaf
characteristics
among
the
others
taxa
of
the
genus,
clearly
separating
these
plants
in
the
morphological
identification
key.
©
2017
Sociedade
Brasileira
de
Farmacognosia.
Published
by
Elsevier
Editora
Ltda.
This
is
an
open
access
article
under
the
CC
BY-NC-ND
license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Introduction
The
Brazilian
Savanna
is
considered
a
hotspot
(priority
area)
for
global
biodiversity
conservation
(Myers
et
al.,
2000);
for
hosting
an
estimated
number
of
837
bird
species,
161
mammal
species,
150
amphibian
species,
and
120
reptile
species
(Silva
and
Bates,
2002);
and
for
high
concentrations
of
endemic
plants
(Mittermeier
et
al.,
2005).
Among
the
great
diversity
of
the
Brazilian
Savanna
endemic
fruits,
we
highlight
those
belonging
to
the
Myrtaceae
family.
The
ecological
importance
of
this
family
in
different
biomes
of
Brazil
has
been
confirmed
in
several
floristic,
phylogenetic,
and
phytoso-
ciological
studies.
According
to
Oliveira-Filho
and
Fontes
(2000),
in
both
the
Brazilian
Savanna
and
Atlantic
Forest,
the
Myrtaceae
family
has
a
great
diversity
of
species,
representing
10–15%
of
the
vegetal
cover
in
these
biomes.
∗Corresponding
author.
E-mail:
joelma.paula@ueg.br
(J.A.
Paula).
One
of
most
comprehensive
surveys
of
the
Brazilian
Myr-
taceae
listed
1726
species
(Berg,
1859).
According
to
Landrum
and
Kawasaki
(1997),
this
family
is
represented
by
23
genera
and
1000
species.
Among
these
plants,
696
species
are
exclusive
to
Brazil
(Arantes,
1997).
In
a
more
recent
study
in
Brazil,
Sobral
et
al.
(2013)
recorded
23
genera
and
approximately
997
species.
In
the
Brazilian
Savanna,
Mendonc¸
a
et
al.
(2007)
listed
14
genera
and
211
species.
Among
the
plant
species
of
the
Myrtaceae
family
previously
cat-
alogued
in
open
areas
of
the
Brazilian
Savanna,
the
Psidium
genus
is
distinguished
by
its
great
economic
potential,
reflecting
its
food
and
pharmacological
uses
(Souza
and
Lorenzi,
2008).
In
Brazil
Psidium
guajava
L.
stands
out
as
the
species
of
greatest
economic
interest
of
the
Psidium
genus.
However,
there
has
been
growing
interest
in
the
“arac¸
azeiros”
(Medina
et
al.,
2011),
which
encompass
several
species
of
Psidium.
In
the
Central
West
region
of
Brazil,
in
addition
to
Psidium
guineense,
the
following
“arac¸
ás”
species
have
also
been
observed:
Psidium
laruotteanum
Cambess.,
Psidium
firmum
O.Berg.,
Psidium
myrsinites
DC.,
Psidium
sartorianum,
and
Psidium
salutare
(Frazon
et
al.,
2009).
With
regard
to
the
medicinal
potential
of
Savanna
“arac¸
ás”,
a
recent
ethnobotanical
survey
has
indicated
the
use
of
leaves
and
http://dx.doi.org/10.1016/j.bjp.2017.03.005
0102-695X/©
2017
Sociedade
Brasileira
de
Farmacognosia.
Published
by
Elsevier
Editora
Ltda.
This
is
an
open
access
article
under
the
CC
BY-NC-ND
license
(http://
creativecommons.org/licenses/by-nc-nd/4.0/).

408
E.F.
Oliveira
et
al.
/
Revista
Brasileira
de
Farmacognosia
27
(2017)
407–413
Legend:
Coordinates
Anápolis
Caldas Novas
Goiás State
Brazil
Anápolis
N
Military
air base
UEG
Caldas Novas
Caldas
Novas
State
Park
20 Km
20 Km
10
1050
50
Fig.
1.
Map
of
Brazil
showing
the
locations
of
Goiás
State,
Anápolis
and
Caldas
Novas
municipalities
and
Psidium
spp.
collection
sites.
Table
1
Geographic
location,
date
of
collection,
geographic
coordinates,
and
voucher
number
of
Psidium
species
collected
in
the
Savanna
of
the
Goiás
state.
Species
Local
Collection
date
Geographic
coordinates
Longitude/latitude/altitude
(m)
Voucher
number
Psidium
firmum
O.Berg.
GSU
08/2013
02/2014
08/2014
16◦2253.977/48◦5641.779 /1098
9211a
Psidium
myrsinites
DC. GSU
06/2013
02/2014
08/2014
16◦2242.058/48◦5640.488 /1099
9212a
AAB
11/2013
06/2014
08/2014
16◦1252.991/48◦5755.868 /1104 9213a
Psidium
laruotteanum
Cambess.
GSU
06/2013
02/2014
08/2014
16◦2246.885/48◦5639.774 /1100
9214a
AAB
11/2013
06/2014
08/2014
16◦1256.088/48◦5756.486 /1114
9215a
SCNSP
08/2013
11/2013
02/2014
06/2014
17◦2812.396/48◦243.604 /982
9218a
Psidium
guineense
Sw.
AAB
11/2013
06/2014
08/2014
16◦1245.507/48◦5723.651 /1104
9217a
GSU,
Campus
Anápolis
of
the
Goiás
State
University;
SCNSP,
Serra
de
Caldas
Novas
State
Park;
AAB,
Anápolis
Air
Base.
aHerbarium
of
the
Goiás
State
University
(HGSU).
shoots
by
residents
of
urban
areas,
settlements
and
“quilombola”
communities
of
the
Goiás
Savanna
for
the
treatment
of
dysenter-
ies
(Campos,
2010).
Data
from
the
scientific
literature
reinforce
the
pharmacological
potential
of
“arac¸
ás”,
since
relevant
biologi-
cal
activities
have
been
attributed
to
the
fruits
and
leaves
of
these
species,
such
as:
antioxidant,
antimicrobial,
antiproliferative
on
human
tumor
cells
and
increased
sleep
induced
by
ketamine
in
mice
(Fauth
et
al.,
2002;
Corrêa
et
al.,
2011;
Medina
et
al.,
2011;
Voss-Rech
et
al.,
2011;
Oliveira
et
al.,
2012;
Patel,
2012).
Several
researchers,
including
Mc
Vaugh
(1968)
and
Barroso
et
al.
(1991),
have
described
the
taxonomic
problems
involv-
ing
Myrtaceae,
stating
that
this
family
is
a
complex
taxonomic
group.
According
to
Costa
(2004),
the
difficulty
of
identifying
Brazilian
Myrtaceae
can
be
attributed
to
the
speciation,
resulting
from
hybridization
and
polyploidy,
and
the
lack
of
morphological
and
anatomical
studies
for
a
better
delimitation
of
the
taxa.
The
Psidium
species,
in
particular,
have
a
great
phenotypic
plasticity
due
to
the
different
environmental
pressures
to
which
they
are
subject,
causing
difficulties
in
the
identification
and
delimitation
of
the
species
(Costa,
2009).
They
are
found
in
diverse
environments
such
as,
semi-desert
and
restingas,
with
calcareous
and
sandy
soils,
among
others
(Brandão
et
al.,
2002).
According
to
Frazon
et
al.
(2009)
in
the
Brazilian
Savanna
there
are
approximately
thirteen
native
or
introduced
Psidium
species,
known
as
“arac¸
azeiros”.
In
relation
for
habit,
these
species
are
char-
acterized
as
sub-shrub
or
tree,
reaching
up
to
five
meters
in
height.

E.F.
Oliveira
et
al.
/
Revista
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de
Farmacognosia
27
(2017)
407–413
409
Fig.
2.
Psidium
spp.
leaves:
(A)
P.
firmum,
adaxial
and
abaxial
surfaces;
(a1)
P.
firmum
diaphanized
leaf
–
brochidodromous
secondary
venation
pattern;
(B)
P.
myrsinites,
adaxial
and
abaxial
surfaces;
(b1)
P.
myrsinites
diaphanized
leaf
–
camptodromous–brochidodromous
secondary
venation
pattern;
(C)
P.
laruotteanum,
adaxial
and
abaxial
surfaces;
(c1)
P.
laruotteanum
diaphanized
leaf
–
camptodromous–brochidodromous
secondary
venation
pattern;
(D)
P.
guineense,
adaxial
and
abaxial
surfaces;
(d1)
P.
guineense
diaphanized
leaf
–
camptodromous–brochidodromous
secondary
venation
pattern.
Among
the
foliar
morphological
characteristics
common
to
these
species,
we
can
highlight
the
simple,
opposite,
usually
crossed,
amphi
or
hypoestomatic
leaves,
with
abundant
tector
trichomes
on
the
abaxial
surface
and
rare
in
the
adaxial
surface,
and
camptodro-
mous
or
brochidodromous
secondary
vein
framework
(Soares-Silva
and
Proenc¸
a,
2008;
Gomes
et
al.,
2009;
Campos,
2010).
Research
concerning
the
aspects
of
leaf
architecture
can
be
as
fundamental
as
the
morphology
of
the
reproductive
organs
used
in
systematic
studies
(Hickey
and
Taylor,
1991).
According
to
Ellis
et
al.
(2009),
the
study
of
leaf
architecture
is
a
relatively
new
approach
that
can
provide
important
information
on
taxonomically
complex
families,
such
as
Myrtaceae.
According
to
Arantes
and
Monteiro
(2002),
despite
the
high
representation
of
Myrtaceae
in
different
biomes,
few
taxonomic
studies
have
been
conducted,
reflecting
the
diversity
of
species
and
taxonomic
complexity
of
these
plants.
The
morphological
studies
of
this
family,
and
especially
of
the
Psidium
genus,
can
contribute
to
ecological
and
phytosociological
studies,
providing
subsidies
to
validate
the
taxonomic
classification
and
recognition
of
new
species
(De-Carvalho,
2008).
Morphological
references
are
extremely
important,
particularly
for
medicinal
plants,
so
that
the
samples
can
be
confirmed
and
to
enable
tests
of
authenticity.
These
tests
are
needed,
because
confu-
sion
between
morphologically
similar
species
is
very
common
and
can
lead
to
the
improper
use
of
a
particular
species.
Thus,
the
leaf
architecture
adds
to
the
set
of
information
that
allows
the
accurate
identification
of
plant
species.
The
aim
of
the
present
study
was
to
describe
the
morphology
and
foliar
venation
of
four
species
of
Psid-
ium
collected
from
a
Savanna
biome
in
Goiás
State,
Brazil,
to
contribute
to
the
identification
and
delineation
of
these
taxa,
and
to
the
quality
control
of
the
plant
material.
Material
and
methods
Leaves
of
Psidium
spp.
were
collected
from
the
Brazilian
Savanna
areas
in
Goiás
State,
in
2013
and
2014,
in
the
cities
of
Anápolis
and
Caldas
Novas
(Fig.
1;
Table
1).
The
botanical
material
comprised
fully
expanded
adult
leaves
collected
below
the
third
node.
For
each
individual,
a
voucher
specimen
was
deposited
in
the
Herbarium
of
Goiás
State
Univer-
sity
–
HGSU
(Table
1),
and
the
next
characteristics
were
recorded
for
each
voucher
specimen:
P.
firmum
O.Berg.:
shrub,
about
90
cm
high;
stem
smooth,
cylindrical,
glabrous,
and
brown;
P.
myrsinites
DC.:
tree,
about
3
m
high;
stem
smooth
cylindrical,
glabrous,
with
irregular
plate
depressions
that
are
detached
from
the
trunk,
slightly
pink
brown
coloration;
P.
laruotteanum
Cambess.:
tree,
about
3
m
high;
stem
with
cracked
rhytidoma
and
sinuous
and
dis-
continuous
crests,
bark
with
dark
grayish
coloration;
P.
guineense
Sw.:
tree,
about
5
m
high;
smooth
cylindrical
stem,
yellowish
gray
coloration.
Leaf
morphological
characterization
was
performed
by
obser-
vations
with
the
naked
eye
using
a
stereoscopic
microscope,
when
necessary,
according
to
Oliveira
and
Akisue
(2003)
and
Ellis
et
al.
(2009).
For
venation
pattern
analyses,
fresh
leaves
were
collected
and
diaphanized
according
to
Kraus
and
Arduin
(1997).
At
least
five
whole
leaves
from
each
specimen
were
placed
in
a
container
with
20%
sodium
hydroxide
for
24
h
to
remove
chlorophyll.
Sub-
sequently,
several
washes
with
water
and
distilled
water
were
performed,
and
the
botanical
materials
were
clarified
using
sodium

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Fig.
3.
Psidium
spp.
leaf
vein
characters.
(A)
P.
firmum,
intersecondary
vein
(arrow);
(a1)
Vein
orders
(until
the
5th
order)
(arrows);
(B)
P.
myrsinites,
irregular
areoles
(square);
(C)
P.
laruotteanum,
freely
ending
veinlets
(FEV)
(arrow);
(D)
Psidium
guineense,
irregular
areoles
and
freely
ending
veinlets
(FEV)
(arrow).
Sv,
secondary
vein;
Tv,
tertiary
vein;
Qn,
quaternary
vein;
Qun,
quinternary
vein;
Ve,
venule;
Mi,
midrib;
Isv,
intersecondary
vein.
hypochlorite.
Subsequently,
dehydration
was
performed
in
a
series
of
70%
(w/w),
90%
(w/w)
and
absolute
ethanol,
incubating
the
plant
material
for
1
h
in
each
solution.
The
leaves
were
stained
with
methylene
blue/xylol
(1:1)
or
safranin/xylol
(1:1).
Each
leaf
was
immersed
in
xylol
and
immediately
transferred
to
a
glass
plate
containing
resin,
mounted
on
two
plates
and
dried.
Anal-
yses
were
performed
using
a
LEICA,
EZ4D
model
stereoscopic
microscope.
The
pattern
venation
registration
was
generated
from
expan-
sions
of
diaphanized
leaves
(using
a
photographic
amplifier
Durst
M601),
and
smaller
networks
of
the
veins
were
recorded
using
a
Olympus
BX40
photomicroscope.
Macro
and
microscopic
charac-
terizations
of
the
leaf
venation
patterns
were
performed
according
to
Cardoso
and
Sajo
(2004,
2006)
and
Ellis
et
al.
(2009).
Results
and
discussion
The
leaves
of
the
Psidium
species
analyzed
in
the
present
study
were
typically
coriaceous,
with
short
petioles
and
a
primary
pin-
nate
type
venation
pattern,
and
characterized
by
only
one
midrib
of
a
higher
gauge.
The
leaf
laminates
have
several
shapes,
and
elliptic
or
obo-
vate
shapes
were
primarily
observed.
The
apex
shapes
were
acute
to
acuminated
in
P.
firmum,
P.
laruotteanum,
and
P.
myrsinites.
P.
guineense
displayed
apex
obtuse
to
mucronate
shapes.
The
base
shapes
were
acute
to
cuneate
in
P.
myrsinites
and
P.
laruotteanum
and
rounded
in
P.
firmum
and
P.
guineense
(Box
1;
Fig.
2).
Trichomes
were
present
on
the
abaxial
surface
of
the
leaf
lamina
of
all
species.
P.
guineense
displayed
trichomes
on
both
surfaces
(Box
1).
The
leaf
midrib
was
more
prominent
on
the
abaxial
surface
in
the
four
species.
P.
firmum
(Fig.
2A)
and
P.
guineense
(Fig.
2D)
pre-
sented
grooved
midribs
on
adaxial
surface
(Box
1).
The
major
secondary
vein
framework
was
the
camptodromous–brochidodromous
mixed
venation
pattern,
followed
by
the
brochidodromous
pattern
(Box
1;
Fig.
2).
The
brochidodromous
venation
pattern
was
observed
in
P.
fir-
mum
(Fig.
2a1),
consistent
with
the
observation
of
Campos
(2010)
for
this
species.
This
venation
pattern
is
formed
when
secon-
daries
join
in
a
series
of
prominent
arches
and
loops
of
secondary
gauge
(Ellis
et
al.,
2009).
According
to
Soares-Silva
and
Proenc¸
a
(2008),
brochidodromous
is
the
typical
venation
pattern
for
Psid-
ium.
Cardoso
and
Sajo
(2006)
reported
that
the
brochidodromous
arches
and
loops
might
be
more
or
less
prominent
depending
on
the
closing
angle
of
the
secondary
veins.
P.
myrsinites,
P.
laruotteanum,
and
P.
guineense
(Fig.
2b1,
c1,
d1)
present
camptodromous–brochidodromous
secondary
vena-
tion
patterns.
This
mixed
venation
pattern
is
characterized
by
secondary
veins
that
anastomose
to
each
other
from
the
leaf
base
in
two
ways:
the
proximal
secondaries
connect
to
superjacent
major
secondaries
via
tertiaries
without
forming
marked
marginal
loops
of
the
secondary
gauge;
in
the
medium
third
of
the
leaf,
the
sec-
ondaries
join
in
a
series
of
prominent
arches
and
loops,
forming
several
closing
angles,
and
in
the
apex,
the
arches
are
concaved
and
uniform
(Ellis
et
al.,
2009).

E.F.
Oliveira
et
al.
/
Revista
Brasileira
de
Farmacognosia
27
(2017)
407–413
411
Box
1:
Leaf
morphological
characters
and
venation
of
four
species
of
Psidium
spp.
Characters/species
P.
firmum
P.
myrsinites
P.
laruotteanum
P.
guineense
Shape
Elliptic
to
oblong
Elliptic
to
obovate
Elliptic
to
obovate
Elliptic
to
obovate
Leaf Apex
Acute
to
acuminate
Acute
to
acuminate
Acute
to
acuminate
Acute
to
mucronate
Base
Rounded
Acute
to
cuneate
Acute
to
cuneate
Rounded
Petiole
Short-petiolate
Short-petiolate
Short-petiolate
Short-petiolate
Indument
Adaxial
surface
Glabrous
Glabrous
Glabrous
Pilose
Abaxial
surface
Pilose
Pilose
Pilose
Pilose
Main
vein
salience
Adaxial
surface
Grooved
Prominent
Prominent
Grooved
Abaxial
surface Prominent Prominent Prominent Prominent
Primary
venation
pattern Pinnate
Pinnate
Pinnate
Pinnate
Secondary
venation
Venation
pattern
Brochidodromous
Camptodromous–
brochidodromous
Camptodromous–
brochidodromous
Camptodromous–
brochidodromous
Number
of
pairs
<13
<13
<13
<10
Salience
adaxial
surface
Flat
Prominent
Flat
Grooved
Salience
abaxial
surface Flat
Flat
Flat
Prominent
Vein
(orders)
Until
the
5th
order
Until
the
5th
order
Until
the
5th
order
Until
the
5th
order
Intersecondary
veins
Present
Present
Present
Present
Intramarginal
veins
Absent
Absent
Absent
Absent
Marginal
ultimate
venation
Fimbrial
Incomplete
loops
Looped
Looped
Collector
vein Distance
from
the
edge
(mm)
<1
1–3
1–3
1–3
Venules
Simple/branching
Branching
Branching
Branching
Branching
Areoles
Present
Present
Present
Present
The
camptodromous–brochidodromous
secondary
venation
pattern
is
common
for
representatives
of
Myrtinae,
Myrciinae
and
Eugeniinae
(Costa
et
al.,
1995;
Farmacopeia
Brasileira,
2002;
Cardoso
and
Sajo,
2004,
2006).
Costa
et
al.
(1995)
described
the
camptodromous–brochidodromous
standard
for
six
of
the
eleven
Eugenia
species
occurring
in
the
sandbank
of
Marica,
Rio
de
Janeiro
State
in
Brazil.
In
the
Brazilian
Pharmacopoeia
IV,
this
secondary
venation
pattern
is
registered
to
P.
guajava
L.
(Farmacopeia
Brasileira,
2002).
Cardoso
and
Sajo
(2004)
identi-
fied
the
camptodromous–brochidodromous
secondary
venation
standard
in
nine
(9)
Eugenia
species
from
the
hydrographic
basin
of
Tibagi
River,
Paraná
State,
Brazil:
Eugenia
arenosa
Mattos,
E.
hamil-
tonii
(Mattos)
Mattos,
E.
handroana
D.
Legrand,
E.
hiemalis
Cambess.,
E.
neoverrucosa
Sobral,
E.
pyriformis
Cambess.,
E.
ramboi
D.
Legrand,
E.
speciosa
Cambess.,
and
E.
uniflora
L.
The
arrangement
of
secondary
veins
relative
to
the
midrib
in
the
leaves
of
the
studied
species
produces
angles
with
divergences
ranging
from
45◦to
75◦.
According
to
Cardoso
and
Sajo
(2004),
the
divergence
angles
ranging
from
45◦to
60◦are
most
striking
in
species
with
mixed
venation
patterns
(camptodromous–brochidodromous),
as
recorded
in
the
present
study
for
P.
laruotteanum,
P.
myrsinites,
and
P.
guineense.
The
diver-
gence
angles
cannot
be
considered
to
be
taxonomic
characters
for
distinguishing
species,
as
in
the
present
study,
all
species
showed
wide
variations
among
the
angles
in
leaves
from
the
same
individ-
ual.
The
amount
of
secondary
vein
pairs
for
each
species
is
shown
in
Box
1.
P.
guineense
(Fig.
2d1)
presents
fewer
secondary
vein
pairs
than
other
species
(8–10
pairs),
thereby
distinguishing
this
species.
All
species
have
veins
until
the
5th
order
(Fig.
3a1).
Note
the
presence
of
intersecondary
veins
in
all
species
(Box
1;
Fig.
3A).
According
to
Ellis
et
al.
(2009),
the
intersecondary
veins
originate
from
the
midrib,
running
parallel
to
the
major
secondary
veins,
as
characterized
by
an
intermediate
gauge
between
the
sec-
ondary
and
tertiary
veins.
In
the
present
study,
no
species
showed
intramarginal
veins,
which
according
to
Cardoso
and
Sajo
(2004),
are
the
veins
formed
between
brochidodromous
arches
and
the
leaf
margin.
Spacing
among
the
secondary
veins
occurred
in
decreasing
order
toward
the
apex.
Irregular
spacing
was
observed
in
P.
firmum,
P.
myrsinites,
and
P.
guineense
(Fig.
2a1,
b1,
d1),
and
regular
spacing
was
observed
in
P.
laruotteanum
(Fig.
2c1).
The
configuration
of
the
highest-order
leaf
veins
in
the
mar-
gin
or
marginal
ultimate
venation
was
absent,
showing
incomplete
loop,
looped,
and
fimbrial
types.
In
the
incomplete
loop
type,
the
marginal
ultimate
vein
recurves
to
form
incomplete
loops
near
the
leaf
margin;
in
the
looped
type,
the
tertiary
veins
anastomose,
forming
a
series
of
arches
near
the
edge;
the
fimbrial
vein
type
occurs
when
close
veins
join
at
the
edge
and
produce
a
continuous
vein
running
along
the
margin
(Cardoso
and
Sajo,
2006;
Ellis
et
al.,
2009).
In
the
present
study,
the
marginal
ultimate
venation
of
the
fim-
brial
type
was
observed
only
in
P.
firmum
(Box
1;
Fig.
4A).
In
P.
myrsinites
(Box
1;
Fig.
4B),
the
incomplete
loop
type
was
observed,
and
in
P.
laruotteanum
(Fig.
4C)
and
P.
guineense
(Fig.
4D)
the
looped
types,
which
have
a
smaller
gauge
closer
to
the
margin,
were
ver-
ified.
According
to
Gomes
et
al.
(2009)
and
Silva
et
al.
(2008),
in
most
surveys
of
Psidium,
the
marginal
ultimate
venation
pattern
is
the
looped
type,
formed
by
complete
arches,
consistent
with
the
results
obtained
in
the
present
study.
The
marginal
ultimate
looped
type
venation
pattern
is
com-
mon
in
most
species
of
the
Myrtaceae
family,
followed
by
fimbrial
and
incomplete
loop
types
(Cardoso
and
Sajo,
2006;
Oliveira
et
al.,
2011).
De-Carvalho
(2008)
registered
the
fimbrial
and
looped
types
in
two
and
seven
species,
respectively,
of
the
nine
species
of
Myrcia
DC.
(Myrtaceae)
studied
in
Federal
District,
Brazil.
Areoles
are
the
smallest
areas
of
the
leaf
tissue
completely
sur-
rounded
by
veins
of
the
fourth
and
fifth
orders.
These
structures
can
have
various
shapes
and
arrangements,
with
or
without
venules
(simple
or
ramified).
The
areoles
are
considered
to
be
regular
(per-
fect)
when
they
have
the
same
shape
and
size;
irregular
(imperfect)
areoles
vary
in
size
and
shape;
and
incomplete
areoles
have
no
limitations
on
the
sides
(Alvarez
et
al.,
2006;
Ellis
et
al.,
2009).
The
results
of
the
present
study
revealed
irregular
areoles
with
freely
ending
veinlets
(FEVs)
of
the
dendritic
ramified
type
for
all
Psidium
species
(Box
1;
Fig.
3B–D).
Costa
et
al.
(1995)
and
Klucking
(1988)
highlighted
that
the
formation
of
incomplete
and
irregularly
shaped
randomly
distributed
areoles
is
a
common
characteristic
in
the
Myrtaceae
family.
In
summary,
the
leaves
of
the
Psidium
species
analyzed
in
the
present
study
are
coriaceous,
short-petiolated
structures
with
a
primary
pinnate
type
venation
pattern.
The
secondary
vena-
tion
pattern
is
brochidodromous,
occurring
more
frequently
the
camptodromous–brochidodromous
mixed
pattern.
The
secondary

412
E.F.
Oliveira
et
al.
/
Revista
Brasileira
de
Farmacognosia
27
(2017)
407–413
Fig.
4.
Psidium
spp.
marginal
ultimate
venation.
(A)
P.
firmum,
marginal
ultimate
venation
of
fimbrial
type
(rectangle);
(B)
P.
myrsinites,
marginal
ultimate
venation
of
incomplete
loop
type
(rectangle);
(C)
P.
laruotteanum,
marginal
ultimate
venation
of
looped
type
(rectangle);
(D)
P.
guineense,
marginal
ultimate
venation
of
looped
type
(rectangle).
Box
2:
Morphological
identification
key
for
Psidium
spp.
1
Shrubby
habit;
brochidodromous
secondary
venation
pattern;
marginal
ultimate
venation
fimbrial
type
P.
firmum
1
Tree
habit;
camptodromous–brochidodromous
secondary
venation
pattern;
marginal
ultimate
venation
looped
type
2
2
Ridged
adaxial
surface
with
prominent
midrib
on
the
abaxial
surface;
pilose
on
both
surfaces;
apex
obtuse
to
mucronate;
base
rounded
P.
guineense
2
Adaxial
surface
with
prominent
midrib;
glabrous
adaxial
surface
and
pilose
abaxial
surface;
apex
acute
to
acuminate;
base
acute
to
cuneate
3
3
Secondary
veins
prominent
on
the
adaxial
surface
and
marginal
ultimate
venation
with
incomplete
loops
P.
myrsinites
3
Secondary
veins
flat
on
the
adaxial
surface
and
marginal
ultimate
venation
looped
P.
laruotteanum
veins
have
different
spacing
patterns,
with
acute
divergence
angles
to
the
midrib,
a
plane
on
the
adaxial
surface
and
are
primarily
prominent
on
the
abaxial
surface.
All
plants
have
intersecondary
veins.
The
marginal
ultimate
venation
of
the
looped
type
appears
in
most
species
and
might
appear
incomplete
and
fimbrial.
The
areo-
las
are
well
developed
veins
of
the
fourth
and
fifth
orders,
and
have
branched
FEVs.
The
different
characters
observed
generated
the
following
iden-
tification
key
for
Psidium
species
(Box
2).
Reflecting
the
small
number
of
species
analyzed
in
the
present
study
with
respect
to
the
biodiversity
reported
for
Brazil,
the
need
for
additional
comprehensive
studies
on
Psidium
species
collected
in
other
Brazilian
biomes
is
evident.
However,
the
morphologi-
cal
characters
and
leaf
architecture
were
useful
for
distinguishing

E.F.
Oliveira
et
al.
/
Revista
Brasileira
de
Farmacognosia
27
(2017)
407–413
413
Psidium
species,
further
confirming
that
diaphanization
is
an
inex-
pensive,
fast
and
conclusive
technique.
Notably,
phenotypic
variation
reflecting
the
influence
of
the
environment
was
not
considered
in
this
analysis,
as
the
sites
where
the
species
were
collected
have
similar
microclimates.
Authors’
contributions
EFO
(Master
student)
contributed
in
collecting
plant
sample
and
identification,
confection
of
herbarium,
running
the
laboratory
work,
analysis
of
the
data
and
drafted
the
paper.
DGB
contributed
in
collecting
plant
sample,
confection
of
herbarium,
running
the
laboratory
work,
analysis
of
the
data
and
drafted
the
paper.
MLS
contributed
in
collecting
plant
sample
and
identification,
confec-
tion
of
herbarium,
analysis
of
the
data
and
to
critical
reading
of
the
manuscript.
MHR
contributed
to
critical
reading
of
the
manuscript.
JAMP
designed
the
study,
supervised
the
laboratory
work
and
con-
tributed
to
critical
reading
of
the
manuscript.
All
the
authors
have
read
the
final
manuscript
and
approved
the
submission.
Conflicts
of
interest
The
authors
declare
no
conflicts
of
interest.
Acknowledgments
The
authors
would
like
to
thank
Jair
Eustáquio
Faria
Júnior
for
assistance
with
material
identification,
Francisco
Junior
Simões
Calac¸
a
for
designing
the
boards,
CAPES
and
Fundac¸
ão
de
Amparo
à
Pesquisa
do
Estado
de
Goiás
for
financial
support.
References
Alvarez,
A.S.,
Potiguara,
R.C.V.,
Santos,
J.U.M.,
2006.
Arquitetura
foliar
de
espécies
de
Eugenia
L.
(Myrtaceae)
da
restinga
de
Algodoal,
Maiandeua,
PA.
Bol.
Mus.
Para.
Emilio
Goeldi
2,
29–36.
Arantes,
A.A.,
(Master
dissertation)
1997.
Florística
da
família
Myrtaceae
Juss.
na
Estac¸
ão
Ecológica
do
Panga,
Uberlândia,
MG.
Unesp,
Rio
Claro,
SP.
Arantes,
A.A.,
Monteiro,
R.,
2002.
A
Família
Myrtaceae
na
Estac¸
ão
Ecológica
do
Panga,
Uberlândia,
Minas
Gerais,
Brasil.
Lundiana
3,
111–127.
Barroso,
G.M.,
Peixoto,
A.L.,
Ichaso,
C.L.F.,
Costa,
C.G.,
Guimarães,
E.F.,
Lima,
H.C.,
1991.
Sistemática
de
angiospermas
do
Brasil.
Universidade
Federal
de
Vic¸
osa,
Imprensa
Universitária,
Vic¸
osa.
Berg,
O.,
1859.
Myrtaceae.
In:
Martius,
C.P.F.
(Ed.),
Flora
brasiliensis
14.
,
pp.
529–656.
Brandão,
M.,
Laca-Buendía,
J.P.,
Macedo,
J.F.,
2002.
Árvores
nativas
e
exóticas
do
Estado
de
Minas
Gerais.
EPAMIG,
Belo
Horizonte.
Campos,
L.Z.O.,
(Master
dissertation)
2010.
Etnobotânica
do
gênero
Psidium
L.
(Myrtaceae)
no
Cerrado
brasileiro.
Brasilia.
Instituto
de
Ciências
Biológicas,
Uni-
versidade
de
Brasília,
Brasília,
71
pp.
Cardoso,
C.M.V.,
Sajo,
M.G.,
2004.
Vascularizac¸
ão
foliar
e
a
identificac¸
ão
de
espécies
de
Eugenia
L.
(Myrtaceae)
da
bacia
hidrográfica
do
Rio
Tibagi,
PR.
Rev.
Bras.
Bot.
27,
47–54.
Cardoso,
C.M.V.,
Sajo,
M.G.,
2006.
Nervac¸
ão
foliar
em
espécies
brasileiras
de
Myr-
taceae
Adans.
Acta
Bot.
Bras.
20,
657–669.
Corrêa,
L.C.,
Santos,
C.A.F.,
Vianello,
F.,
Lima,
G.P.P.,
2011.
Antioxidant
content
in
guava
(Psidium
guajava)
and
arac¸
á
(Psidium
spp.)
germplasm
from
different
Brazilian
regions.
Plant
Genet.
Resour.
9,
384–391.
Costa,
I.R.,
(Master
dissertation)
2004.
Estudos
cromossômicos
em
espécies
de
Myr-
taceae
Juss.
no
sudeste
do
Brasil.
Instituto
de
Biologia,
Universidade
Estadual
de
Campinas,
Campinas.
Costa,
I.R.,
(Ph.D.
thesis)
2009.
Estudos
evolutivos
em
Myrtaceae:
aspectos
cito-
taxonômicos
e
filogenéticos
em
Myrteae,
enfatizando
Psidium
e
gêneros
relacionados.
Instituto
de
Biologia,
Universidade
Estadual
de
Campinas,
Cam-
pinas,
235
pp.
Costa,
C.G.,
Machado,
R.D.,
Fontenelle,
J.B.,
1995.
Sistema
vascular
em
folhas
de
Eugenia
L.
(Myrtaceae).
Bol.
Herb.
Bradea.
6,
345–356.
De-Carvalho,
P.S.,
(Master
dissertation)
2008.
Myrcia
DC.
ex
Guill.
(sec¸
ão
Myrcia,
Myrtaceae)
no
Distrito
Federal,
Brasil.
Universidade
de
Brasília,
Brasilia,
68
pp.
Ellis,
B.,
Daly,
D.C.,
Hickey,
L.J.,
Johnson,
K.R.,
Mitchell,
J.D.,
Wilf,
P.,
Wing,
S.L.,
2009.
Manual
of
Leaf
Architecture.
Cornell
University
Press,
Ithaca.
Farmacopeia
Brasileira,
2002.
Monografia
Goiabeira,
4th
ed.
parte
2.
fasc.
4,
Atheneu,
São
Paulo.
Fauth,
S.,
Campos,
A.R.,
Silveira,
E.R.,
Rao,
V.S.,
2002.
Efeitos
de
óleos
essenciais
de
plantas
no
tempo
de
sono
induzido
por
cetamina
em
camundongos.
Rev.
Bras.
Farmacogn.
12,
112–113.
Frazon,
R.C.,
Campos,
L.Z.O.,
Proenc¸
a,
C.E.B.,
Sousa-Silva,
J.C.,
2009.
Arac¸
ás
do
gênero
Psidium:
principais
espécies,
ocorrência,
descric¸
ão
e
usos.
Embrapa
Cerrados,
Brasília
(Série
Documentos,
Embrapa).
Gomes,
S.M.A.,
Somavilla,
N.S.D.N.,
Gomes-Bezerra,
K.M.,
Miranda,
S.C.,
De-Carvalho,
P.S.,
Graciano
Ribeiro,
D.,
2009.
Anatomia
foliar
de
espécies
de
Myrtaceae:
contribuic¸
ões
à
taxonomia
e
filogenia.
Acta
Bot.
Bras.
23,
223–238.
Hickey,
L.J.,
Taylor,
D.W.,
1991.
The
leaf
architecture
of
Ticodendron
and
the
appli-
cation
of
foliar
characters
in
discerning
its
relationships.
Ann.
Mo.
Bot.
Gard.
78,
105–130.
Klucking,
E.P.,
1988.
Leaf
Venation
Patterns.
Myrtaceae.
J.
Cramer,
Stuttgart.
Kraus,
J.E.,
Arduin,
M.,
1997.
Manual
básico
de
métodos
em
morfologia
vegetal.
EDUR,
Seropédica.
Landrum,
L.R.,
Kawasaki,
M.L.,
1997.
The
genera
of
Myrtaceae
in
Brasil:
an
illustrated
synoptic
treatment
and
identification
keys.
Brittonia
49,
508–536.
Mc
Vaugh,
R.,
1968.
The
genera
of
American
Myrtaceae:
an
interim
report.
Taxon
17,
354–418.
Medina,
A.L.,
Haas,
L.I.R.,
Chaves,
F.C.,
Salvador,
M.,
Zambiazi,
R.C.,
Silva,
W.P.,
Nora,
L.,
Rombaldi,
C.V.,
2011.
Arac¸
á
(Psidium
cattleianum
Sabine)
fruit
extracts
with
antioxidant
and
antimicrobial
activities
and
antiproliferative
effect
on
human
cancer
cells.
Food
Chem.
128,
916–922.
Mendonc¸
a,
R.C.,
Filgueiras,
T.S.,
Fagg,
C.W.,
2007.
Análise
Florística
da
Chapada
dos
Veadeiros.
In:
Felfili,
J.M.,
Rezende,
A.V.,
Silva,
J.R.
(Eds.),
Biogeografia
do
Bioma
Cerrado:
Vegetac¸
ão
e
Solos
da
Chapada
dos
Veadeiros.
Editora
Universidade
de
Brasília/Finatec,
Brasília,
pp.
119–237.
Mittermeier,
R.A.,
Robles,
P.,
Hoffmann,
M.,
Pilgrim,
J.,
Brooks,
T.,
Mittermeier,
C.G.,
Lamoreux,
J.,
Fonseca,
G.B.,
2005.
Hotspots
Revisited:
Earth’s
Biologically
Richest
and
Most
Endangered
Ecoregions.
Conservation
International,
Mexico
City.
Myers,
N.,
Mittermeier,
R.A.,
Mittermeier,
C.G.,
Da
Fonseca,
G.A.B.,
Kent,
J.,
2000.
Biodiversity
hotspots
for
conservation
priorities.
Nature
403,
853–858.
Oliveira,
F.,
Akisue,
G.,
2003.
Fundamentos
de
farmacobotânica,
2nd
ed.
Atheneu,
São
Paulo.
Oliveira-Filho,
A.T.,
Fontes,
M.A.L.,
2000.
Patterns
of
floristic
differentiation
among
Atlantic
Forests
in
Southeastern
Brazil
and
the
influence
of
climate.
Biotropica
32,
793–810.
Oliveira,
M.I.U.,
Funch,
L.S.,
Santos,
F.A.R.,
Landrum,
L.R.,
2011.
Aplicac¸
ão
de
carac-
teres
morfoanatômicos
foliares
na
taxonomia
de
Campomanesia
Ruiz
&
Pavón
(Myrtaceae).
Acta
Bot.
Bras.
25,
455–465.
Oliveira,
V.B.,
Yamada,
L.T.,
Fagg,
C.W.,
Brandão,
M.G.L.,
2012.
Native
foods
from
Brazilian
diversity
as
a
source
of
bioactive
compounds.
Food
Res.
Int.
48,
170–179.
Patel,
S.,
2012.
Exotic
tropical
plant
Psidium
cattleianum:
a
review
on
prospects
and
threats.
Rev.
Environ.
Sci.
Biotechnol.
11,
243–248.
Silva,
F.A.M.,
Assad,
E.D.,
Evangelista,
B.A.,
2008.
Caracterizac¸
ão
climática
do
Bioma
Cerrado.
In:
Sano,
S.M.,
Almeida,
S.P.,
Ribeiro,
J.F.
(Eds.),
Cerrado:
Ecologia
e
Flora.
Embrapa,
Planaltina,
pp.
69–88.
Silva,
J.M.C.,
Bates,
J.M.,
2002.
Biogeographic
patterns
and
conservation
in
the
South
American
Cerrado:
a
tropical
savanna
hotspot.
BioScience
52,
225–233.
Soares-Silva,
L.H.,
Proenc¸
a,
C.E.B.,
2008.
A
new
species
of
Psidium
L.
(Myrtaceae)
from
southern
Brazil.
Bot.
J.
Linn.
Soc.
158,
51–54.
Sobral,
M.,
Proenc¸
a,
C.,
Souza,
M.,
Mazine,
F.,
Lucas,
E.,
2013.
Myrtaceae.
In:
Lista
de
Espécies
da
Flora
do
Brasil.
Jardim
Botânico
do
Rio
de
Janeiro,
http://floradobrasil.jbrj.gov.br/jabot/floradobrasil/FB171/
(accessed
December
2013).
Souza,
V.C.,
Lorenzi,
H.,
2008.
Botânica
sistemática:
guia
ilustrado
para
identificac¸
ão
das
famílias
de
Angiospermas
da
flora
brasileira,
baseado
em
APG
II.
Instituto
Plantarum,
Nova
Odessa.
Voss-Rech,
D.,
Klein,
C.S.,
Techio,
V.H.,
Scheuermann,
G.N.,
Rech,
G.,
Fiorentin,
L.,
2011.
Antibacterial
activity
of
vegetal
extracts
against
serovars
of
Salmonella.
Cienc.
Rural
41,
314–320.