Leaf structure of Bromelia and its significance for the evolution of Bromelioideae (Bromeliaceae)

ORIGINAL ARTICLE

Leaf structure of Bromelia and its significance for the evolutionof Bromelioideae (Bromeliaceae)

Raquel Fernandes Monteiro • Rafaela Campostrini Forzza •

Andre Mantovani

Received: 5 April 2010 / Accepted: 28 January 2011 / Published online: 10 March 2011

� Springer-Verlag 2011

Abstract This study investigated the leaf structure of

Bromelia and its importance for understanding the evolution

of Bromelioideae (Bromeliaceae). Because of the scarcity of

informative molecular characters in Bromeliaceae, this

study evaluates the relevance of anatomical characters for

the taxonomy and phylogeny of Bromelia and the subfamily

Bromelioideae. Anatomical studies in monocots have shown

that the combined application of anatomical and external

morphological characters from leaf structure can improve

the taxonomic delimitation of species, genera and subgen-

era, and is very informative for inferring phylogenies. The

current study analyzed the leaves of 27 species of Bromelia

and found that the most important characters for the sys-

tematics of this group are the occurrence of a water storage

hypodermis, the number of stalk cells of peltate scales, the

presence of a ribbed abaxial surface, the occurrence of pal-

isade parenchyma on the adaxial side, the shape of the cells

that surround the air lacunae, the presence of raphides and

secretory channels, and the occurrence of fibrous extensions

on the bundle sheath on minor veins. Combining our results

with those described for the family, we made a list of the

anatomical characters that can be used in phylogenetic

studies of Bromelioideae.

Keywords Foliar anatomy � Neotropics � Monocots �Systematics � Taxonomy

Anatomical characters can be useful in the phylogenetic

analysis of Bromeliaceae, especially considering that

informative molecular characters are scarce. For instance,

only 6% of the characters obtained from plastid region

sequenced markers of 48 species in 24 genera of Bromeli-

aceae were found to be informative (Schulte et al. 2009; see

also Faria 2006; Sousa et al. 2007). Anatomical features are

considered potentially useful to support taxonomic cir-

cumscriptions within the monocots as a whole (e.g., Simon

2007; Henderson and Stevenson 2007; Simpson and Burton

2006; Furness and Rudall 2000; Stevenson et al. 2000;

Tomlinson and Fisher 2000) and particularly within Bro-

meliaceae (e.g., Tomlinson 1969; Braga 1977; Sousa and

Neves 1996; Sajo et al. 1998; Aoyoma and Sajo 2003;

Arruda & Costa 2003; Scatena and Segecin 2005; Sousa

et al. 2005; Horres et al. 2007; Proenca and Sajo 2007,

2008). Within the subfamily Bromelioideae, with unclear

generic delimitation (Schulte et al. 2009, Benzing 2000),

combined anatomical and morphological characters have

already proved useful to solve taxonomic problems (e.g.,

Ramirez 1996; Brown 2000; Coffani-Nunes 2004; Sousa

2004; Sousa et al. 2007; Almeida et al. 2009).

Bromelia, a basally diverging genus of Bromelioideae

(Givinish et al. 2007; Schulte et al. 2009), includes 56

species (Luther 2008) grouped into three subgenera (Mez

1891): Distiacanthus, Karatas and Bromelia. It occurs

from central Mexico to southern Argentina, with two

centers of diversity: one in Central America extending to

the southern Andes and the other in the Brazilian Shield,

mainly in Cerrado vegetation, where the majority of its

species are found, most of them in dry or more markedly

seasonal regions (Smith and Downs 1979). Species of

Bromelia can be recognized by their leaves with curved

spines along the margins, sheaths covered with fine linear

scales, fleshy petals united into a tube by the filaments, lack

of petal appendages and flattened naked seeds (Smith and

Downs 1979). The phylogenetic study with the genus

R. F. Monteiro (&) � R. C. Forzza � A. Mantovani

Jardim Botanico do Rio de Janeiro, Rua Pacheco Leao 915,

Rio de Janeiro, RJ 22460-030, Brazil

e-mail: [email protected]

123

Plant Syst Evol (2011) 293:53–64

DOI 10.1007/s00606-011-0426-2

(Monteiro 2009) showed the occurrence of only three

synapomorphies for the genus: the fuzzy indumentum on

the abaxial surface of the leaf sheath, the oblong ovary and

the sclerenchymatous hypodermis with four layers on the

abaxial surface. Micromorphological studies with Bromelia

provide general anatomical descriptions of a few species

(Tomlinson 1969; Schmidt and Brown 2004), without

evaluating their potential for phylogenetic systematics

(Horres et al. 2007).

Despite the importance of the anatomical features in the

systematics of Bromeliaceae, there is no information

available for many of its genera. In this study we examined

the leaves of 27 species of Bromelia species with the aim of

pointing out characters that can be useful on phylogenetic

analysis of the genus and the subfamily.

Materials and methods

Twenty-seven Bromelia species were studied in an attempt

to cover all the different habitats and the distribution of the

genus (Table 1). Expanded leaves were collected in the

field and preserved in ethanol 70%. Vouchers were

deposited at the Herbarium of the Jardim Botanico do Rio

de Janeiro (RB). In addition, dry leaves from exsiccatae

were analyzed after rehydration and subsequent treatment.

Both freehand sections and resin embedding were

prepared.

Cross sections of the middle third of the leaf blade were

made freehand. One or two cross sections were replicated

for each specimen. The freehand cuts were cleared in 50%

sodium hypochlorite, stained with Safranin and Astra blue

Table 1 Species of Bromelia analyzed with their respective collection sites and voucher specimens

Taxon Provenance Voucher specimen

Subgenus Distiacanthus

B. morreniana (Regel) Mez Brazil, Amazonas G.T.Prance 16484 (INPA)

B. scarlatina (Hort. ex Herincq) E. Morren Peru, Loreto T. Plowman 7032 (F)

Subgenus Karatas

B. auriculata L.B.Sm. Brazil, Bahia B.R.Silva 1452 (JBRJ hort)

B. grandiflora Mez Brazil, Goias E.M.C.Leme 2200 (RB)

B. karatas L. Costa Rica, Puntarenas E.Bello 2903 (F)

B. lagopus Mez Brazil, Goias R.F.Monteiro 131 (RB)

B. lindevaldae Leme & E. Esteves Brazil, Goias B.R.Silva 1223 (RB)

B. macedoi L.B.Sm. Brazil, Goias R.F.Monteiro 126 (RB)

B. minima Leme & E. Esteves Brazil, Goias B.R.Silva 1253 (RB)

B. aff. tubulosa L.B.Sm. Brazil, Espırito Santo B.R.Silva 1387 (JBRJ hort), R.F.Monteiro 280 (RB)

B. tubulosa L.B.Sm. Brazil, Para M.J.Pires 1661 (INPA)

B. villosa Mez Brazil, Goias B.R.Silva 1234 (RB)

Subgenus Bromelia

B. antiacantha Bertol. Brazil, Rio de Janeiro B.R.Silva s.n. (JBRJ hort), R.F.Monteiro 263 (RB)

B. arenaria Ule Brazil, Bahia R.C.Forzza 3834 (RB)

B. balansae Mez Brazil, without locality E.M.C.Leme 2869 (RB)

B. binotii E. Morren ex Mez Brazil, Espırito Santo R.F.Monteiro 265 (RB)

B. chrysantha Jacq. Venezuela, Lara A.Fernandez 1473 (F)

B. goyazensis Mez Brazil, Goias B.R.Silva 1458 (RB)

B. hemispherica Lam. Mexico, Chiapas R.M.Laughlin 859 (F)

B. hieronymii Mez Argentina, Santiago del Estero V.Marunak 395 (RB)

B. horstii Rauh Brazil, Mato Grosso do Sul C.Fleming s.n. (RB)

B. interior L.B.Sm. Brazil, Goias R.F.Monteiro 122 (RB)

B. irwinii L.B.Sm. Brazil, Goias H.S.Irwinii 33100 (F)

B. laciniosa Mart. ex Schult. & Schult. f. Brazil, Minas Gerais B.R.Silva 1409 (JBRJ hort)

B. pinguin L. United States, Saint Croix Roadside 342 (F)

B. reversacantha Mez Brazil, Goias R.F.Monteiro 130 (RB)

B. serra Grisb. Brazil, Minas Gerais R.F.Monteiro 233 (RB)

Acronyms of herbaria are according to Thiers B (continuously updated)

54 R. F. Monteiro et al.

123

(Bukatsch 1972), and mounted under a coverslip in 50%

glycerin for light microscope viewing.

For embedding in resin, cross sections from the same

region of the leaf measuring approximately 2 cm2 were

initially fixed in glutaraldehyde, dehydrated in an increas-

ing alcohol series and included in resin hydroxyethylmet-

acrylate. Cross sections of approximately 3 lm thick were

obtained using a Spencer rotary microtome and stained

with Toluidine blue O (O’Brien and McCully 1981).

Digital photomicrographs were taken using a Coolsnap

digital camera coupled with an Olympus BX-50 optical

microscope. Cell and tissue classifications followed the

nomenclature proposed by Tomlinson (1969).

Results

All leaves are covered by a one-layered epidermis of thick-

walled cells usually with the inner periclinal walls thicker

than the outer ones and cells with reduced lumens (Fig. 1d, e).

The cuticle is thin in all species (Fig. 1d), and Bromelia

karatas Fig. (2d) also has a thick layer outside of the epi-

dermal cells at the adaxial surface, which may be wax and

needs to be investigated further. In B. grandiflora and

B. horstii, some epidermal cells are papillated (Fig. 1f). The

adaxial surface is smooth in all leaves (Figs. 1a, b, c, g; 3a,

b), and the abaxial one is smooth to undulating in B. binotii,

B. chrysantha, B. grandiflora, B. horstii, B. lagopus,

Fig. 1 Leaf anatomy of

Bromelia, optical microscopy.

a Bromelia reversacantha;

b B. binotii; c B. goyazensis;

d B. morreniana, adaxial

epidermis and hypodermis

details. Arrow, epidermal cell

with a reduced lumen.

e B. morreniana abaxial

epidermis and hypodermis

details. Arrow, epidermal cell

with a reduced lumen.

f B. grandiflora, epidermis and

hypodermis details. Arrow,

papillae. g B. arenaria. WSwater storage tissue, CPchlorenchyma with palisade

cells, AL air-lacunae

parenchyma, ST sclerified

hypodermis. Scale bar 100 lm

in a, b, c and g; 50 lm in f,30 lm in d and e

Leaf structure of Bromelia 55

123

B. lindevaldae, B. macedoi, B. minima, B. morreniana,

B. scarlatina and B. aff. tubulosa (Figs. 1b, 3a, b), and

strongly ribbed in the remaining species (Fig. 1a, c, g).

All leaves are hypostomatic with the stomata at the

same level of the epidermal cells in B. aff. tubulosa,

B. hemispherica, B. irwinii, B. lindevaldae, B. reversacantha

and B. tubulosa (Fig. 2a), and sunken in the others

(Fig. 2b, c). The guard cells show thick walls, reduced

lumens and cuticular outer ledges closing the ostiole. Sub-

stomatal chambers may be partially occluded because of the

enlargement of the subsidiary cells beneath the guard cells

(Fig. 2c).

Peltate scales are attached to the epidermis by a stalk

consisting of two cells in almost all examined specimens

(Fig. 2e). In B. serra and B. horstii the stalk is formed by

three cells (Fig. 2f). The scales occur only on the abaxial

surface of B. binotii, B. horstii, B. grandiflora, B. karatas,

B. minima, B. villosa and B. aff. tubulosa, and in both sur-

faces of the other species. The scale shield is large in

diameter in B. scarlatina (Fig. 3a) and small in the others

(Fig. 2c).

Adjacent to the epidermis there is a hypodermis of one

to five layers of sclerified cells on both leaf sides of all

examined species. In B. horstii these cells appear only on

the abaxial surface. The sclerified hypodermal cells have

slightly thickened walls (Fig. 2e) in B. grandiflora,

B. interior, B. laciniosa, B. macedoi, B. reveresacantha,

B. scarlatina and B. aff. tubulosa, and greatly thickened

walls in the other species (Fig. 1d).

Internally to the sclerified hypodermis there is a water

storage tissue formed by rounded parenchymatous cells in

B. auriculata, B. binotii, B. chrysantha, B. hemispherica,

B. grandiflora, B. interior, B. macedoi, B. morreniana,

B. reversacantha, B. scarlatina, B. tubulosa and B. aff.



a Bromelia tubulosa, detail of

the stomata; b B. laciniosa,details of the stoma;

c B. arenaria, peltate scale and

stomata; d Bromelia karatas,

layer of wax above the cuticle;

e B. laciniosa, peltate scales at

abaxial face; f B. horstii, peltate

scale. WS water storage tissue,

AL air-lacunae parenchyma, Schsubstomatal chamber, STsclerified hypodermis. Scalebar = 100 lm in d, 50 lm in a,

b, c and e, 30 lm in f


123

tubulosa (Figs. 1a, b; 3a, b), and by anticlinally elongated

cells in B. goyazensis and B. arenaria (Fig. 1c, g). In most

of the species, this tissue is restricted to the adaxial surface

(Fig. 1a, b, c, g), but in B. morreniana it is found in the

abaxial side and in B. aff. tubulosa and B. scarlatina in

both surfaces. The transition between the water storage

tissue and the chlorenchyma is abrupt in most species

(Fig. 1a, b, c, g), although it is subtle in B. auriculata, B.

morreniana, B. scarlatina and B. aff. tubulosa (Fig. 3a, b).

The chlorenchyma is formed by rounded cells (Fig. 3a,

b), by palisade cells (Fig. 1a, b, c g) or by armed cells that

delimit air spaces forming air gaps. The armed cells are

long, delimiting large air spaces (Figs. 3c, 2c), or short,

delimiting small ones (Figs. 1a, c, 3d). They occur inter-

calating with vascular bundles and occasionally extending

to the abaxial leaf side. The leaves of B. antiacantha, B.

auriculata, B. chrysantha, B. hemispherica, B. hieronymii,

B. karatas, B. morreniana, B. pinguin, B. scarlatina, B.

tubulosa and B. aff. tubulosa do not possess palisade cells.

In all species examined, the vascular bundles are collat-

eral and arranged in a single series with the larger ones

usually alternating with the smaller ones (Figs. 1a, b, c, g; 3a,

b; 4a). The large bundles are completely surrounded by

sclerified cells (Figs. 1b, c, g; 3a, b; 4c), but in B. hieronymii,

B. interior, B. karatas, B. macedoi, B. reversacantha,

B. aff. tubulosa and B. villosa, they are restricted to the xylem

and phloem regions (Figs. 1a, 4a). Externally to these scle-

rified cells, a sheath of isodiametric parenchymatous cells

surrounds all the vascular bundles. In B. lindevaldae,

B. hemispherica and B. irwinii, the parenchymal cells are

radially elongated (Fig. 4d).

The small-diameter vascular bundles have caps of

sclerified cells near the xylem and phloem that form

small extensions towards the surface in B. arenaria,

B. auriculata, B. balansae, B. chrysantha, B. goyazensis,

B. hemispherica, B. horstii, B. interior, B. irwinii, B. karatas,

B. lagopus, B. lindevaldae, B. macedoi, B. serra and B. aff.

tubulosa (Figs. 1c, 4c), and large extensions in

B. hieronymii (Fig. 4b). In the other species the small

vascular bundles are covered by caps that are wider than

long (Figs. 1b, 4a).

Idioblasts with raphides were observed in B. grandiflora,

B. goyazensis, B. horstii, B. lagopus, B. macedoi, B. minima,

B. serra and B. villosa (Fig. 4e). Scattered secreting

channels appear in the chlorenchyma of B. arenaria,

B. grandiflora, B. goyazensis, B. horstii, B. laciniosas,

B. macedoi, B. reversacantha and B. serra (Fig. 4f). Extra-

fascicular fibers were not found in the genus.

Tables 2, 3 and 4 summarize the results obtained for all

species examined here.

Discussion

The leaf structure of all species studied here is in accor-

dance with the general pattern described for Bromeliaceae



a Bromelia scarlatina; b B.morreniana, water storage

parenchyma islands; c B.binotii, details of the air-lacunae

parenchyma; d B. minima,

details of the air-lacunae

parenchyma. WS water storage

tissue, AL air-lacunae

parenchyma. Scalebar = 100 lm in a and b,

50 lm in c and d


123

by Tomlinson (1969) and corresponds to type IV of the

classification for the subfamily Bromelioideae suggested

by Horres et al. (2007), which recognizes four groups of

species based on leaf anatomy. In this type of leaves, the

vascular bundles are completely immersed in the chloren-

chyma, the extra-vascular fibers are absent, and both the

adaxial water storage tissue and air gaps in the air lacunae

are developed. Only B. aff. tubulosa, B. scarlatina and B.

morreniana do not completely agree with this type as their

water storage tissue is formed by small groups of paren-

chymatous cells adjacent to the abaxial surface.

The epidermal cells are U-shaped with the inner peri-

clinal wall thickened in all Bromelia studied here, cor-

roborating the description of Tomlinson (1969) for other

representatives of the family. Epidermal cells with uniform

thickness along the entire wall and a large lumen seem to

be a rare character within the Bromeliaceae as it does not

appear in any representatives studied by Aoyoma and Sajo

(2003), Scatena and Segecin (2005), Proenca and Sajo

(2007), Almeida (2006) and Vargens (2008).

The sunken stomata often hidden by the scales, as

observed here for the Bromelia species, is probably asso-

ciated with a reduction in transpiration rates, as suggested

by Tomlinson (1969). A sunken stomata was observed for

other Bromeliaceae from xeric environments (Pita 1997;

Forzza 2001, Arruda and Costa 2003; Proenca and Sajo

2007), although it can also appear in some epiphytic

Aechmea, Billbergia, Hohenbergia, Neoregelia, Quesnelia

and Wittrockia of humid habitats (Sajo et al. 1998; Sousa

et al. 2005; Almeida 2006; Faria 2006; Vargens 2008). In

Bromeliaceae, the stomata are rarely superficial, although

this was observed in Vriesea species of xeric environments

by Arruda and Costa (2003). Thus, the stomata position

appears to be more related to the phylogeny of the group



a Bromelia villosa, details of the

vascular bundles;

b B. hieronymii, details of the

vascular bundles; c B. serra,

details of the vascular bundles;

d B. irwinii, details of the

vascular bundles; e B. balansae,

raphides; f B. laciniosa,secreting channel. R raphides,

SC secreting channel. Scalebar = 100 lm in a, b, c and d;

50 lm in f; 30 lm in e


123

than to the environmental conditions where the species are

found (Scatena and Segecin 2005; Proenca and Sajo 2007).

The specialized trichomes of Bromeliaceae that cover

the blade and the leaf sheath are commonly associated with

the absorption of water and nutrients (e.g., Benzing 2000),

especially in Tillandsioideae. In Bromelioideae, this func-

tional role is commonly assumed by the scales covering

only the sheath, not the limb (Benzing 1970). Possibly, in

terrestrial species with functional roots (e.g., Bromelia,

Puya, Dyckia, Encholirium, Navia), the scales only protect

against water loss, reflecting the light and providing water

repellence (Pita 1997; Forzza 2001; Pierce et al. 2001). If

in Bromelia the leaf scale is a little or not absorbent, it is

possible to infer that trichomes that absorb water and

nutrients appeared more than once during the evolutionary

history of the Bromeliaceae, although it needs to be con-

firmed by the analysis of the phylogenetic trees provided

by Givinish et al. (2007) and Schulte et al. (2005, 2009).

Moreover, considering Puya as a sister group to other

Bromelioideae and the genus Bromelia as basal in the

subfamily, it is possible to say that absorbent trichomes

appeared after the Bromelia cladogenesis, where it dif-

fered. Schulte et al. (2009) also stated that, in Bromelioi-

deae, the first emergence of absorbent scales is directly

linked to the appearance of tanks formed by leaf sheaths

and blades.

Also regarding the trichomes, the number of stalk cells

as well as the number, arrangement and shape of the shield

cells are features with taxonomic importance in the sub-

families and genera of Bromeliaceae (Forzza 2001).

According to Tomlinson (1969), the number of stalk cells

is a diagnostic feature for the genera, but not the number

and shape of shield cells, which vary in a single leaf,

limiting their use for species differentiation. In

Table 2 Character list based on Bromeliaceae leaf anatomy observed in studied species

Species/

character

Adaxial surface,

contour

Abaxial surface,

contour

Epidermal

cells of adaxial

surface

Epidermal

cells of abaxial

surface

Number

of stalk

cells

Peltate scales

on adaxial

surface

Stomata position

in relation

to the epidermis

Smooth to

undulating

Ribbed Smooth to

undulating

Ribbed Inner periclinal

wall thickeness

Inner periclinal

wall thickeness

Superficial Sunken

B.aff. tubulosa ? – ? – ? ? 2 – ? –

B. antiacantha ? – – ? ? ? 2 ? – ?

B. arenaria ? – – ? ? ? 2 ? – ?

B. auriculata ? – – ? ? ? 2 ? – ?

B. balansae ? – – ? ? ? 2 ? – ?

B. binotii ? – ? – ? ? 2 – – ?

B. chrysantha ? – ? – ? ? 2 ? – ?

B. goyazensis ? – – ? ? ? 2 ? – ?

B. grandiflora ? – ? – ? ? 2 – – ?

B. hemispherica ? – – ? ? ? 2 ? ? ?

B. horstii ? – ? – ? ? 3 – – ?

B. interior ? – – ? ? ? 2 ? – ?

B. irwinii ? – – ? ? ? 2 ? ? –

B. karatas ? – – ? ? ? 2 – – ?

B. laciniosa ? – – ? ? ? 2 ? – ?

B. lagopus ? – ? – ? ? 2 ? – ?

B. lindevaldae ? – ? – ? ? 2 ? ? –

B. macedoi ? – ? – ? ? 2 ? – ?

B. minima ? – ? – ? ? 2 – – ?

B. morreniana ? – ? – ? ? 2 ? – ?

B. pinguin ? – – ? ? ? 2 ? – ?

B. reversacantha ? – – ? ? ? 2 ? ? –

B. scarlatina ? – ? – ? ? 2 ? – ?

B. serra ? – – ? ? ? 3 ? – ?

B. tubulosa ? – – ? ? ? 2 ? ? –

B. villosa ? – – ? ? ? 2 – – ?

? Presence, - absence


123

Bromelioideae, stalks with more than two cells probably

arose after the cladogenesis of the ancestral Bromelia,

constituting a synapomorphy for the other genera of the

subfamily, with a reversal in the Quesnelia/Billbergia clade

and in some species of Aechmea (Vargens 2008; Almeida

et al. 2009).

The presence of ribs in the leaf surface has been used by

Tomlinson (1969), Pita (1997) and Forzza (2001) as a

character for species distinction within Bromeliaceae.

These ribs, usually associated with intercoastal areas and

the presence of stomata and trichomes, were observed only

on the abaxial surface of the Bromelia species studied here

and can be a potential characteristic for the genus. In con-

trast, the adaxial surface is always smooth or undulating.

Similarly to epidermal cells, in most of the species, the

sclerified hypodermis is formed by cells with thickened

walls. This thickening that also occurs in the epidermal

tissue may be associated with limiting transpiration, aiding

the survival of individuals subjected to constant water

stress and offering mechanical support and protection

against herbivores (Magalhaes et al. 2005; Scatena and

Segecin 2005). Based on the phylogenetic hypotheses for

Bromeliaceae (Givinish et al. 2007; Schulte et al. 2005,

2009), it is possible to infer that a sclerified hypodermis

appeared and was lost in several clades independently. We

can also say that the presence of a sclerified hypodermis

does not seem to be related to water availability, as many

species living in humid environments possess this tissue in

Table 3 Character list based on Bromeliaceae leaf anatomy observed in studied species

Species/character Adaxial

sclerenchymatous

hypodermis

Abaxial

sclerenchymatous

hypodermis

Adaxial water

storage tissue, size

Transition

between the

water storage

tissue and the

chlorenchyma

Adaxial

water

storage

tissue

Air-lacunae

parenchyma,

cells form

Number

of cell

layers

Cell wall

thickness

Number

of cell

layers

Cell wall

thickness

Up to 1/3

of the

blade

Larger than

1/3 of the

blade

Abrupt Subtle Cell shape

B. aff. tubulosa ? 1 Greatly ? 1 Greatly ? – – ? Rounded Short arms

B. antiacantha ? 2 G/S ? 3 Greatly ? – ? – Elongated Long arms

B. arenaria ? 2 Slightly ? 3 Greatly – ? ? – Elongated Long arms

B. auriculata ? 3 Greatly ? 4 Greatly ? – – ? Rounded Short arms

B. balansae ? 2 Slightly ? 3 G/S ? – ? – Elongated Short arms

B. binotii ? 2 Greatly ? 2 Greatly ? – ? – Rounded Long arms

B. chrysantha ? 1 Greatly ? 5 G/S – ? ? – Rounded Long arms

B. goyazensis ? 2 Slightly ? 3 Slightly – ? ? – Elongated Short arms

B. grandiflora ? 2 G/S ? 3 G/S ? – ? – Rounded Long arms

B. hemispherica ? 3 Greatly ? 4 Greatly – ? ? – Rounded Long arms

B. horstii – 0 – ? 2 Greatly – ? ? – Elongated Long arms

B. interior ? 2 Slightly ? 3 Slightly ? – ? – Rounded Short arms

B. irwinii ? 3 G/S ? 4 G/S – ? ? – Elongated Long arms

B. karatas ? 3 Greatly ? 5 Greatly – ? ? – Elongated Long arms

B. laciniosa ? 1 Slightly ? 2 G/S – ? ? – Elongated Short arms

B. lagopus ? 2 Greatly ? 2 Greatly ? – ? – Elongated Short arms

B. lindevaldae ? 3 G/S ? 4 Greatly – ? ? – Elongated Short arms

B. macedoi ? 2 Slightly ? 3 Greatly ? – ? – Rounded Short arms

B. minima ? 2 Greatly ? 2 Greatly – ? ? – Elongated Short arms

B. morreniana ? 1 Greatly ? 2 Greatly – – – ? Rounded Long arms

B. pinguin ? 2 Greatly ? 3 Greatly – ? ? – Elongated Long arms

B. reversacantha ? 2 Slightly ? 2 Greatly ? – ? – Rounded Short arms

B. scarlatina ? 1 Slightly ? 2 Greatly ? – – ? Rounded Long arms

B. serra ? 3 Greatly ? 4 Greatly ? – ? – Elongated Short arms

B. tubulosa ? � Greatly ? 2 Greatly ? – ? – Rounded Short arms

B. villosa ? 2 Slightly ? 2 Greatly ? – ? – Elongated Short arms

? Presence, - absence, G/S greatly and slightly


123

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123

their leaves (e.g., Sousa et al. 2005, Almeida 2006; Faria

2006), and others in non-xeric settings also do (e.g., Forzza

2001). The number of layers of hypodermis cells is variable

inside the genera in Bromeliaceae (Tomlinson 1969; Sousa

et al. 2005; Almeida 2006; Faria 2006; Vargens 2008).

Near the sclerified hypodermis a multilayered paren-

chymatous tissue provides water storage, making the leaves

more or less succulent (Mantovani 1999; Proenca and Sajo

2004; Scatena and Segecin 2005). The cells of this tissue

can be isodiametric or vary from isodiametric to anticlinal

elongated in the inner layers, as observed in B. antiacan-

tha. Occasionally, these cells show shrinkage, possibly due

to water loss (Krauss 1949), and, because they are located

just above the photosynthetic layers, Brighigna et al.

(1984) argued that this region protects the chlorenchyma

from high light levels in order to optimize the photosyn-

thesis and reduce the risk of photoinhibition.

An abaxial water storage tissue was only observed in B.

aff. tubulosa, B. morreniana and B. scarlatina, and seems

to be unusual in other basal genera of Bromelioideae (Terry

et al. 1997; Sajo et al. 1998; Schulte et al. 2005; Givinish

et al. 2004; 2007). Apparently, this is a derived character

that appeared several times in different genera of the

subfamily.

Chlorenchyma endowed with palisade cells, as observed

here for the Bromelia, is an uncommon feature in the

family, and up to the present has been found only in Puya,

Brocchinia, Pitcairnia, Encholirum and Acanthostachys

(Tomlinson 1969; Downs 1974; Forzza 2001). Considering

the phylogenetic hypotheses for the family (Givinish et al.

2007; Schulte et al. 2005; 2009), we can infer that palisade

tissue had appeared several times independently. However,

the presence of this feature in both Puya and Bromelia

suggests that it can occur in other Bromelioideae and had

appeared again in Acanthostachys.

The aeration channels delimited by armed parenchy-

matous cells, as observed here for Bromelia, are common

in Bromeliaceae (Aoyoma and Sajo 2003; Scatena and

Segecin 2005; Proenca and Sajo 2007). According to

Tomlinson (1969) and Gilmartin et al. (1989), these

channels facilitate leaf ventilation. In these channels, the

cell shape is constant for the same leaf region in the same

species and can be useful to distinguish the Bromelia

(Braga 1977), because the arm size of the cells varies from

species to species.

Regarding the secreting channels of Bromeliaceae,

Tomlinson (1969) describes them as formed by cells with a

mucilaginous content in Aechmea and Billbergia. How-

ever, for Bromelia species they are reported here for the

first time. As recent studies do not corroborate the presence

of such channels in Aechmea and Billbergia (Aoyoma and

Sajo 2003; Sousa et al. 2005; Vargens 2008), it is possible

to say that they are rare in the family and are an important

taxonomic characteristic for differentiating Bromelia from

the other Bromelioideae. Although raphides are common

within the family, it is possible to separate species of

Aechmea (Aoyoma and Sajo 2003; Sousa et al. 2005) from

those of Bromelia by the presence or lack of them.

The vascular bundles are arranged in a single series, as

in other species of the family (Tomlinson 1969; Downs

1974; Pita 1997; Forzza 2001; Arruda and Costa 2003;

Aoyoma and Sajo 2003, Scatena and Segecin 2005 ; Sousa

et al. 2005; Proenca and Sajo 2007), and can be completely

surrounded by sclerified cells or present fiber caps adjacent

to xylem and phloem. This variation was also found in

several other groups of the family (e.g., Tomlinson 1969;

Aoyoma and Sajo 2003) and seems to have arisen inde-

pendently several times during the evolutionary history of

the group. However, this character seems important for

supporting small clades within Bromelia (Monteiro 2009)

and may also be useful in the taxonomy of other genera.

The small vascular bundles often show small fiber

extensions towards the abaxial surface in Bromelia species.

Large vascular fiber extensions as well as their complete

absence are rare in the genus. This feature may have been

lost several times independently during the evolution of the

family because these fibers do not appear in many groups

of different sides of the phylogenetic tree, like all studied

Tillandsioideae (Tomlinson 1969; Arruda and Costa 2003;

Segecin and Scatena 2005; Proenca and Sajo 2007;

Givinish et al. 2007). Each vascular bundle is also sur-

rounded by an outer parenchymatous sheath whose cells,

with slightly thickened walls and a wide lumen, are in a

slightly radiating arrangement in three Bromelia. The

radiating arrangement of the parenchymatous sheath was

also reported for other genera, such as Aechmea, Brew-

caria, Brocchinia, Billbergia, Encholirium, Fernesea,

Hohenbergia, Pitcairnia, Portea, Puya and Quesnelia

(Sajo et al. 1998; Forzza 2001; Aoyoma and Sajo 2003;

Sousa et al. 2005; Almeida 2006). Considering the known

phylogenetic relationships among these genera (Terry et al.

1997; Givinish et al. 2004, 2007; Barfuss et al. 2005,

Schulte et al. 2005), we can assume that this is a highly

homoplastic feature.

Fibers not associated with vascular bundles are unusual

in Bromeliaceae, occurring only in derived Bromelioideae

genera, like Aechmea, Canistrum and Quesnelia, and being

rare in Tillandsioideae (Tomlinson 1969; Proenca and Sajo

2007). Taking into account the basal position of Bromelia

(Givinish et al. 2004, 2007; Schulte et al. 2005, 2009), the

absence of extra-fascicular fibers in these genera and in

Puya seems to be a plesiomorphic state within the

subfamily.

The four layers of abaxial sclerenchymatous hypodermis

are the only anatomic synapomorphy for Bromelia

(Monteiro 2009). The results presented here show that,


123

although we can recognize groups of species of Bromelia

using leaf anatomy, no characteristic is unique to any of the

three subgenera proposed for Bromelia by Mez (1891).

Based on the present results as well as on those previ-

ously described, the leaf anatomy seems to provide features

useful in the phylogeny and the delimitation of the Bro-

meliaceae taxa. Therefore, micromorphology must be

emphasized alongside the macromorphology in the sys-

tematics of Bromeliaceae and of monocots over molecular

characters to produce better results for phylogenies as a

rewarding approach, as recently shown by Horres et al.

(2007) and Faria (2006).

Acknowledgments The authors would like to thank Anna Karla

Venda for assistance with the cuttings, Renato de Mello-Silva, Andrea

Ferreira da Costa, Cassia Monica Sakuragui, Daniela Zappi and Graca

Sajo for the criticisms and suggestions, and Conselho Nacional de

Desenvolvimento Cientıfico e Tecnologico (CNPq) and Coordenad-

oria de Aperfeicoamento do Pessoal de Nıvel Superior (CAPES) for

fellowships.

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Leaf structure of Bromelia and its significance for the evolution of Bromelioideae (Bromeliaceae)

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