International Research Journal of Plant Science (ISSN: 2141-5447) Vol. 2(2) pp.

xxx-xxx, March, 2011

Available online http://www.interesjournals.org/IRJPS
Copyright © 2011 International Research Journals

Full length Research Paper

Comparative analysis of stomatal characters in eight

wild atlas pistachio populations (Pistacia atlantica
Desf.; Anacardiaceae)
Safia Belhadj,1* Arezki Derridj,2 Alfonso Moriana,3 Maria Del Carmen Gijon,4 Jean-Phillipe
Mevy5 and Thierry Gauquelin 5
1
* Département d’Agropastoralisme, Université « Ziane Achour » de Djelfa, B.P.3117 Poste Ain Chih, Djelfa 17000,
Algeria.
2
Laboratoire de Biosystématique Végétale, Faculté des Sciences Biologiques et Agronomiques, Université “Mouloud
Mammeri”, Tizi-Ouzou 15000, Algeria. E-mail: aderridj@yahoo.fr. 3 EUITA, University of Seville, Crta de Utrera Km, 1,
41013 Sevilla, Spain. E-mail: amoriana@us.es. 4 CMA El Chaparillo, Consejería de Agricultura, Junta de Castilla-La
Mancha, Crta. de Porzuna Km 3.5, 13071 Ciudad Real, Spain. E-mail: cgijon@jccm.es. 5 Institut Méditerranéen
d'Ecologie et de Paléoécologie. UMR CNRS 6116, Université de Provence, Case 421 – Av. Escadrille Normandie
Niemen, 13397, Marseille Cedex 20, France. E-mail: Jean-Philippe.Mevy@univ-provence.fr, thierry.gauquelin@univ-
provence.fr.
Accepted 21 March, 2011

Little is known about the morphological variations or differences in stomatal characteristics of Pistacia
species. How these variations may be related to the environment is also unknown. This paper describes
the diverse stomatal morphology recorded among the Algerian representatives of Pistacia atlantica Desf.
(Atlas pistachio). Leaf Samples of eight Algerian sites of Pistacia atlantica were characterised according
to their stomatal type, distribution and position in the epidermis, shape, size and density by the mean of
light microscopy and scanning electron microscopy. The leaves are amphistomatic in all the sites.
However the abaxial stomatal densities are much higher. The mean number of stomata per square
millimetre was 30.5 on the adaxial face and 308.5 on the abaxial face. The stomata were elliptical and
slightly sunkun in the epidermis, of the actinocytic or anomocytic types with a mean length of 29.1µm
and a mean width of 19.5µm. However, the variations in the aperture of stomata did not reveal a clear
pattern in response to the ecological gradient tested, suggesting that it is a genotypic character. Only,
the stomatal density showed agreement with the environmental conditions for the areas where the trees
were grown. Especially up to 900 m in altitude ecological conditions abaxial leaf stomata density
decreases significantly compared to that of lowland habitats. The best discriminating traits recorded in
this study were the stomata size and the stomatal density. These characteristics may afford an initial
screening method for classifying P. atlantica in terms of drought resistance.

Keywords: environment, physiology, Pistacia atlantica, SEM, size, shape, stomata.

INTRODUCTION

In North Africa, more than 100,000 hectares are lost to
desertification processes every year (Le Houérou, 1995;
Gauquelin et al., 1999). The Sahara and the semi-arid
areas of North Africa, along with its steppe-land, are
*Corresponding author: Tel: (213) 27900203(04). Fax: (213)
27900201. E-mail: belhadjsafia@yahoo.fr.
home to flora of great interest (FAO, 2007). The
preservation of local floral diversity is required if
reforestation programmes are to be improved and
appropriate ecotypes selected for different areas. The
genus Pistacia ,including P. lentiscus L., P. terebinthus
L., P. atlantica Desf. and P. vera L., is widely present in
North Africa however Pistacia atlantica is the most
common (Zohary, 1952; Quézel and Santa, 1963,
Monjauze, 1980). These trees are important in forest
regions because they provide protection from wind and
water erosion and contribute to soil stability and
conservation (Esmail-Pour, 2001). On the basis of leaf
and fruit characteristics and of eco-geographical
distribution, P. atlantica is subdivided into four varieties:
(1) latifolia, (2) kurdica, (3) kabulica and (4) atlantica, with
the latter being the one occurring in Algeria (Zohary,
1952; Alyafi, 1979; Monjauze, 1980).
Pistacia atlantica (Atlas pistachio) is an endemic
perennial woody plant in North Africa (Ozenda, 1983;
White, 1986; Quézel and Médail, 2003). It used to be
widely distributed across arid and semi-arid regions; it
occurs as isolated scattered trees or in dense populations
at a range of altitude from less than 200m up to 1200m
(Monjauze, 1980; Ozenda, 1983; Quézel and Médail,
2003). Atlas pistachio is one of the main constructive
species of arid and semi-arid Mediterranean forest
ecosystems and plays a very important role in retaining
ecological stability preventing desertification. It is a
drought-resistant plant able to grow in harsh conditions in
which few tree species can be grown and established
(Mirzaie-Nodoushan and Arefi, 2001); its large rooting
system renders it suitable for reforestation programmes
in the semi- arid and arid areas. Nowadays, intense,
germplasm erosion is underway due to human activities
and intensive grazing, within its wide range of distribution
and especially in Algeria (Quézel and Santa, 1963;
Monjauze, 1980; Belhadj, 1999, 2001, 2002; Quézel and
Médail, 2003). Actually, this species is being conserved
and propagated in its potential area however; population
differences in morphological and physiological properties
have not been well established in this species. Despite
the numerous studies on the genus, limited data are
available on the physiological characteristics of this
species underlying its drought resistance mechanisms.
Using morphological characters, such as those related to
leaves and fruits (Belhadj et al., 2008), to leaf epidermis
(Belhadj et al., 2007a) and to pollen grains (Belhadj et al.,
2007b) we showed that the studied Atlas pistachio
populations could be divided into different groups. Leaf
micromorphology and especially the stomata
characteristics could potentially be used as discriminating
characters as well. To our knowledge, few investigations
have been performed on the stomatal characteristics of
Pistacia species. Although great contributions have been
made to the description of leaf micromorphology (Alyafi,
1979; Lin et al., 1984; El-Oqlah, 1996; Çaglar and Tekin,
1999; Özeker and Misirli, 2001; Ait-Said et al. 2004; Kadi-
Bennane et al., 2005; Smail-Saadoun, 2005), stomata of
only a few species have been examined with the light
microscopy and fewer with the scanning electron
microscopy. Atlas pistachio is not well known
systematically and ecologically, and it is quite difficult to
make distinction between P. atlantica varieties by the
classical leaf morphological traits, since several
specimens may have intermediary characters (Zohary,
1952, 1987; Yaltirik, 1967). Pistacia atlantica is known to
have a higher adaxial leaf stomatal density than P.
khinjuk Stocks or P. vera (Çaglar and Tekin, 1999;
Özeker and Misirli, 2001), and a lower abaxial one
compared to P. terebinthus. No conclusions regarding the
relationship with drought resistance or ecosystem type
have been drawn. Stomatal characteristics may therefore
be good descriptors of the environmental conditions
reigning in different ecosystems. The aim of this study
was (i), to access the impact of contrasting local climatic
conditions on P. atlantica leaf stomata traits with
emphasis on the physiological mechanisms involved (ii),
to discriminate phenotypic or genotypic traits that could
serve to some extent as references toward infra-specific
classification of P. atlantica.
MATERIALS AND METHODS
Plant material: Pistacia atlantica leaves used for this study were
collected from eight different sites around Algeria. Specimens were
selected such as to cover a wide climatic variation, and each site is
characterized by its Mediterranean bioclimatic type using Q3, the
classical Emberger’s quotient (Table 1). For each site, the following
voucher specimens were kept in the herbarium of the University of
Provence, Marseille (France): Guerrara, (Mar-2007-PA-G);
Berriane, (Mar-2007-PA-Be); Brezina, (Mar-2007-PA-Br); Elhamel,
(Mar-2007-PA-E); Messaad, (Mar-2007-PA-M); Ain Oussara, (Mar-
2007-PA-Ao); Aflou, (Mar-2007-PA-Af) and finally Oued Safene,
(Mar-2007-PA-Os). Other leaf samples were kept as well in the
herbarium of the University of Djelfa (Algeria).
Light microscopy: In each area, five randomly selected samples of
fully expanded leaflets were taken from different leaves, from five
healthy trees (a total of 200 leaflets). The samples were allowed to
dry under laboratory environmental conditions until use. After
several months they were remoisten in distilled water for 15 to 30
minutes and, when dry, the leaf impression technique was applied
to view stomata (Banon et al., 2004). A thin layer of clear nail
varnish was painted onto both adaxial and abaxial leaf surfaces and
left for 5 to 10 min. A strip of transparent sticky tape (sellotape) was
placed over the dried varnish and pressure applied to obtain an
imprint. The sellotape with its imprint was peeled from the leaflets
and placed onto a glass microscope slide. Replicas were examined
under an optical microscope (Leica D.M.L.S, Germany). The length
and width of ten stomata per leaf were measured on the abaxial
face (a total of 2000 measurements). In addition, the stomata in ten
adaxial and ten abaxial areas of each leaf were counted (a total of
4000 measurements).
Scanning electron microscopy: Others leaves were soaked in
distilled water or either cleaned with ethanol (90%) in order to
remove external particles and dust then, standard procedure was
followed for SEM (Belhadj et al., 2007a). Two specimens from each
site were examined. A section of 5 mm2 of the dry leaf surface (both
adaxial and abaxial surfaces) was fixed on a labelled stub. The
samples were coated with carbon and scanned in a JEOL. JSM-
6360 LV-Japan Microscope. Micromorphological observations
included stomata shape and type description, distribution and
position in the epidermis. SEM pictures were digitally recorded in
different magnifications. Stomatal terminology was based on the
classification proposed by Metcalfe and Chalk (1950), Baranova
(1972, 1983, 1987, 1992), Wilkinson (1979) as well as the studies
of Prabhakar (2004) and Carpenter (2005).
Statistical analysis: The data were subjected to ANOVA and the
mean separation was made with the Tukey (HSD) test to determine
whether the differences among and within populations were
 Table 1. Main climatic features of the selected experimental sites.

Sites Latitude Longitude Altitude M. m. An. Rain Q3 Climate type

(m) (ºC) (ºC) (mm)
Guerrara (G) 32º48’N 4º29’E 107 42.5 4.7 67.2 6 Saharian mild
Berriane (Be) 32º51’N 3º46’E 764 36 2.3 119.7 12.2 Saharian fresh
Brezina (Br) 33º6’N 1º15’E 900 35.1 0.3 230.2 22.5 Arid fresh
Elhamel (E) 35º9’N 4º4’E 730 38.7 4 247.9 24.5 Arid mild
Messaad (M) 34º11’N 3º36’E 780 35.4 1.3 250.8 25.1 Arid fresh
Ain Oussara (Ao) 35º25’N 2º50’E 820 35.8 0.5 238.2 25.9 Arid fresh
Aflou (Af) 34º9’N 2º5’E 930 33.4 -0.05 342 35 Semi arid cold
Oued Safene (Os) 34º55’N 0º48’E 1100 32.7 1.8 462.9 51.2 Semi arid fresh

Source of climatic data: National Meteorology Office of Algeria.

Q3: Emberger quotient, M: Mean of the maxima temperatures of the warmest month, m: Mean of the minima temperatures of the
coldest month.

significant or not using Statistix analytical software (version 1.0,
1996).
RESULTS
Stomata distribution and position in the epidermis: The
leaves were amphistomatic in all the sites; the stomata
were present both on the abaxial (Figures 1A, 1B) and
the adaxial face (Figures 2A-D, 2F). On the adaxial face,
they were located exclusively along and near the midribs,
while on the abaxial surface they were widely spread and
sunken (Berriane, Messaad, Aflou and Elhamel) slightly
sunken below the epidermis (Guerrara, Brezina, Elhamel,
Ain Oussara and Oued Safene) to level (Guerrara and
Brezina) with the epidermis (Table 2; Figures 1B, 1D, 2C-
E, 3).
Stomata type, shape and size: Four different stomata
types were recorded in our study for P. atlantica leaves
(published data in Belhadj et al., 2007a). The most
frequently observed were the actinocytic (Figure 1C) and
the anomocytic types. Nonetheless, the laterocytic and
paracytic types were recorded as well. In the actinocytic
type, the stomata were surrounded by a circle of radiating
cells while in the anomocytic type, the stoma were
surrounded by a limited number of cells that are
indistinguishable in size, shape, or form from those of the
remainder of the epidermis (Metcalfe and Chalk 1950,
Baranova 1983). The guard cells were surrounded by a
ring of subsidiary cells in the laterocytic type while the
paracytic type was characterized by one or two lateral
subsidiary cells oriented parallel to the guard cells
(Baranova 1983, Carpenter 2005). Two to four types
were recorded within a same population in this study. In
Aflou (Af) site only the actinocytic and the anomocytic
types were recorded. The paracytic type was not
recorded in the Elhamel (E) and Guerrara (G) sites
whereas in the Berriane (Be), Brezina (Br), Messaad (M),
Ain oussara (Ao) and Oued safene (Os) sites the four
types were recorded (Table 2).
The different sites varied significantly in the dimensions
of their stomata on the abaxial face (Table 2). Their
dimensions varied between 23.9 µm (E site) to 32.6 µm
(Os site) in length with a mean value of 29.1 µm. For the
width, the values were lower than those recorded for the
length. The mean value was about 19.5 µm and the least
value (15.7 µm) was recorded for the E site whereas the
highest (21.2 µm) for the Os site. The stomata were
longer than larger in all the sites with a mean ratio L/W
around 1.52. The highest value (1.56) was recorded in
the E site whereas the lowest value (1.46) was recorded
in the Ao and Af sites. The stomata were elliptical in all
the sites except for the Ao and Be sites where they
appeared somehow circular (Table 2, Figures 3C, 3F).
Variation among and within populations expressed by the
coefficient of variation was more or less high for the
stomatal dimensions. The coefficients of variation were
high among populations especially for the width (18.6%)
and the length (17.9%). The variation within populations
was important both in length and width as well for the
ratio. The highest values were recorded, for the length in
M (17.6%) and Be (17.2%) sites whereas for the width,
the E (18.9%) and Be (18.1%) sites had the highest
variation. The lowest values were recorded in G site both
for length and width (respectively, 11% and 12%).
Concerning the ratio, the E site had the highest value of
variation (20.9%); the lowest (14.7%) was recorded in the
G site.
The statistical analysis showed highly significant
differences among and within populations both in length
and width except for Br site (Table 2). The Os site had
the longest stomata, followed by G site (no significant
differences with the latter). Messaad site was near the
previous group but significantly lower than Os. Aflou and
Br sites (no significant differences between both of them)
were lower than the first ones but without significant
difference with M. Bellow 30 µm was Ao site though
without significant difference with Af and Br. Finally, Be
and E sites which are significantly the lowest. Variation in
width was less appreciable. The widest stomata (>20 µm)
 Table 2. Stomata characteristics of P. atlantica leaves from different sampling sites. Data are means ± SD, range (Min-Max) and coefficient of
variation (%).

stomatal density Stomata position

Site Stomata length (L) Stomata width (W) L/W ratio (no.mm -2 ) in the epidermis
(µm) (µm) Abaxial Adaxial
Guerrara (G) 31.5***ab ± 3.7 20.6***ab ± 2.5 1.55*ab ± 0.23 318.5***bc ± 47.3 44.7***ab ± 17.2 Level with to
22.0-43.0 (11.0) 12.0-30.0 (12.0) 1.0-2.33 (14.7) 194.6-457.6 (14.9) 0.0-94.7 (38.5) slightly sunken
Berriane (Be) 24.8***e ± 4.3 16.6***c ± 3.0 1.52NSabc ± 0.24 329.6***ab ± 58.6 41.9***b ± 19.4 Sunken
14.0-37.0 (17.2) 10.0-27.0 (18.1) 0.93-2.31 (16.2) 78.9-583.9 (17.8) 0.0-99.9 (46.3)
Brézina (Br) 29.9NScd ± 3.8 19.7NSb ± 3.0 1.54NSabc ± 0.23 281***f ± 41.3 42.6NSb ± 12.6 Level with to
20.0-40.0 (12.8) 10.0-30.0 (15.1) 1.07-2.50 (15.03) 194.6-431.3 (14.7) 0.0-78.9 (29.7) slightly sunken
Elhamel (E) 23.9***e ± 4.0 15.7***c ± 2.9 1.56***a ± 0.33 307***cd ± 51.1 7.4NSe ± 8.2 Slightly sunken to
13.0-35.0 (16.8) 8.0-28.0 (18.9) 0.75-2.87 (20.9) 205.1-541.8 (16.7) 0.0-36.8 (110.6) sunken
Messad (M) 30.9***bc ± 5.4 20.8***a ± 3.3 1.50***abc ± 0.25 338.1***a ± 83.6 6.6NSe ± 7.7 Sunken
18.0-47.0 (17.6) 12.0-32.0 (15.9) 0.90-2.34 (16.6) 10.5-589.1 (24.8) 0.0-36.8 (116.7)
Ain oussara (Ao) 29.1***d ± 3.4 20.4***ab ± 3.3 1.46NSc ± 0.23 309.9***cd ± 47.8 16.3**d ± 12.4 Slightly sunken
22.0-43.0 (11.5) 14.0-35.0 (16.1) 0.88-2.28 (16.1) 31.6-431.3 (15.4) 0.0-57.9 (76.5)
Aflou (Af) 30.2***cd ± 4.2 20.8***a ± 2.7 1.46NSbc ± 0.22 283.5***ef ± 49.7 35.4***c ± 14.7 Sunken
22.0-44.0 (13.9) 12.0-30.0 (12.8) 1.00-2.53 (14.8) 157.8-473.4 (17.6) 0.0-78.9 (41.6)
Oued safène (Os) 32.6***a ± 5.3 21.2***a ± 3.4 1.55**a ± 0.24 299.8***de ± 67.2 49.3***a ± 18.9 Slightly sunken
20.0-46.0 (16.4) 14.0-31.0 (16.2) 1.07-2.25 (15.3) 168.3-478.7 (22.4) 0.0-105.2 (38.7)
Mean 29.1*** ± 5.23 19.5*** ± 3.62 1.52*** ± 0.25 308.5*** ± 60.33 30.5*** ± 21.9 Slightly sunken
13.0-47.0 (17.9) 8.0-35.0 (18.6) 0.75-2.87 (16.5) 10.5-589.1 (19.6) 0.0-105.2 (71.9)
a,b,c, d, e
Mean separation within columns, by Tukey test (p<0.001). Values with same letters are not significantly different. .
*: significant at p<0.05; **: significant at p<0.01; ***: significant at p<0.001; NS: not significant

were noticed in the Os, M, Af, G and Ao sites (no
significant difference), followed by those of Br (no
significant difference to Ao and G, although
different to the other members of the preceding
group). The narrowest stomata were found in the
Be and E sites (no significant difference between
the two).
Stomatal density: Stomatal densities differed
statistically among and within sites. The variations
were less apreciable in the adaxial face, with no
significant differences in Br, E and M sites (Table
2). Sites M (338.1 no/mm2) and Be (329.6
2
no/mm ) showed the highest abaxial stomatal
densities. The lowest values were recorded in the
Br (281 no/mm2) and Af (283.5 no/mm2) sites. The
mean value was about 308.5 no/mm2. Concerning
the adaxial stomatal densities, the values varied
from 6.6 no/mm2 in the M site to 49.3 no/mm2 in
the Os site whereas the mean value was about
30.5 no/mm2 (Table 2). Variations among and
within populations for the stomatal densities were
high especially for the adaxial ones. The
coefficient of variation varied from 14.7% (Br) to
24.8% (M), with a mean value of 19.6% in the
abaxial face. From 29.7% (Br) to 116.7% (M) and
a mean value of 71.9% in the adaxial face (Table
2).
The statistical analysis showed highly significant
differences among the populations for the
stomatal densities. Thought the populations were
separated onto three groups for the abaxial
density and onto four groups for the adaxial one
(Table 2). In the abaxial face, the number of
stomata varied significantly among the
populations with the M site being the first followed
by Be site but not significant from each other.
 Figure 1. SEM photographs of stomata distribution in leaves of P. atlantica on the abaxial leaf side, from the Aflou site. (A)
Stomata appearance and density (x 400), (B) stomata appearance at 25% leaf plane inclination (x 650), (C, D) higher
magnification of stomata, (C) Actinocytic type (x 1200) and (D) sunkun stoma (x 1100), respectively.

These were followed by the G site (no significant
difference with the Be site, but significantly different to the
M site) and the Ao and E sites (both of them significantly
different from Be and M sites but no significant from the
G site). The Os and Af sites (significantly not different
from each other) were near the Br site (no significant
difference) which was significantly different from the rest
(Table 2). In the abaxial face, sites Os and G were the
first (no significant difference between the two) followed
by the Be and the Br sites (no significant difference
between the two and the previous). The Af site was near
the Ao site (both of them significantly different from each
other and from the previous sites). Finally, the E and M
sites were not different from each other but significantly
different from the rest of the sites (Table 2).
DISCUSSION
The results described in this report deal with the effect of
such ecological parameters as temperature, precipitation
and altitude (Table 1) on the micromorphological features
on P. atlantica leaf stomata.
The over all leaf stomata distribution was not affected by
the local climatic conditions investigated. Indeed, our
observations showed that the leaves were
amphistomatic, this is in accordance with the studies of
Lin et al. (1984) and Özeker and Misirli (2001).
Amphistomaty is a common feature in xeric habitats
(Fahn, 1967; Yiotis et al., 2006), and is regarded as
adaptation designed to raise maximum leaf conductance
to CO2 (Bondada and Oosterhuis, 2000). Concerning the
adaxial leaf surface, the stomata were located only near
the major and minor veins in accordance with the studies
of Lin et al. (1984) while they were spread all over the
whole surface as described by Özeker and Misirli (2001).
Besides in our study, the stomata were slightly sunken in
the most of the sites whereas they were level with the
epidermis in Alyafi’s (1979) and Lin et al. (1984) studies.
In most mesomorphic plants the stomata are level with or
very slightly sunken below or raised above the rest of the
epidermis, but in some xeromorphic plants they are often
sunken to a considerable degree (Stace, 1965). Our
results provide evidence that stomata position according
to epidermis surface level reflects adaptations to the
native habitats where the plant grows.
Investigating stomata size and shape we came to the
conclusion that the stomata were longer than wide with a
constant ratio L/W along the ecological gradient tested.
Hence the lack of variation in stomata aperture suggests
that this trait is rather genotypic than adaptative. To this
 Figure 2. SEM photographs of stomata distribution in leaves of P. atlantica on the adaxial leaf side. (A, B) stomata along the
midrib, (A) Berriane (x 300), (B) Brezina (x 400); (C, D, E, F) higher magnifications of stomata, (C) Berriane (x 1100), (D)
Elhamel (x 900), (E) Berriane (x 1600), (F) Berriane (x 950).

stand point, the stomata size recorded in this study are
not similar to those reported by other authors for the
same species: they were shorter and wider for Özeker
and Mirisli (2001), longer and wider for Ait-Said et al.
(2004) (Table 3). With regard to the stomata shape, they
were described as elliptical or orbicular depending to the
local conditions. From the resurrection plant Reaumuria
soongorica (Pall.) Maxim it has been shown that upon
rewatering, stomata were ring-shaped (Liu et al., 2007)
suggesting that elliptical and orbicular forms may be
considered as xeromorphic characters.
Stomatal characters are widely used in the
characterisation of plants belonging to different ecotypes.
These characteristics correlate negatively with one
another in most of the species studied so far. No strict
relationship exists between stomatal size or density and
the regular availability of water. Nonetheless, adaxial and
abaxial leaf stomatal characters are commonly related as
reported in Eucalyptus spp. (Ngugi et al., 2004) and in
Glycine max L. (soybean) (Gitz et al., 2005). In addition
other environmental factors such as soil salinity, CO2
concentration, frost and disease tolerance can affect
these leaf characteristics (Roselli et al., 1989; Botti et al.,
1998; Sharma et al., 2001; Lawson et al., 2002). Several
authors have used stomatal size and density to
characterise lines growing at different sites, usually with
 Table 3. Stomata characteristics of Pistacia atlantica. Comparison of data from different sources.

Author Abaxial Stomata Abaxial Stomata Abaxial Stomatal Adaxial Stomatal Stomata type Stomata Origin
length (µm) width (µm) density (no.mm-2) density (no.mm -2) position
Alyafi (1979) / / / / / Level with the /
epidermis
Lin et al. (1984) / / 569 ± 142 84 ± 24 Actinocytic Level with the California (USA)
epidermis
Özeker and Misirli 18.9 36.3 81.78 439.8 / / Manisa Yunt
(2001) mountain (Turkey)

Ait-Said et al. (2004) 42.4 34.9 282.9 0 / / Ain oussara (Algeria)

40.4 32.7 303.4 0 Messaad (Algeria)
24.3 14.9 427.4 0 Taissa (Algeria)
Kadi-Bennane et al. / / 282.9 0 Anomocytic perigenous / Ain oussara (Algeria)
(2005) 303.4 0 Anomocytic mesoperigenous Messaad (Algeria)
427.4 0 Anisocytic mesoperigenous Taissa (Algeria)
Paracytic mesoperigenous
Paracytic mesogenous
Smail-Saadoun / / / 0 Anomocytic perigenous / Algeria
(2005) Anomocytic mesoperigenous
Anisocytic mesoperigenous
Paracytic mesoperigenous
Paracytic mesogenous
Our results 29.1 ± 5.2 19.5 ± 3.6 308.5 ± 60.3 30.5 ± 21.9 Actinocytic Level with to Algeria
(mean values) Anomocytic slightly sunken
Paracytic in the epidermis
Laterocytic

respect to drought resistance (XueJun and XinShi,
2000; Balok and Hilaire, 2002), but also with the
amount of shade (Gamage et al., 2003) or the
NO2 content of the air (Siegwolf et al., 2001). In
the natural dry lands environment, trees are
frequently subjected to seasonal water stress,
which limits their growth and development. Trees
differ markedly in their ability to withstand
desiccation and have evolved complex
mechanisms to cope with various environmental
stresses including drought. Plants respond to
drought through changes in morphological,
physiological, biochemical and metabolic
processes (Elfadl and Luukkanen, 2006).
A careful examination of the climatic conditions of
the investigated sites (Table 1) showed that
precipitations increase with elevation while
maximum temperature decreases. Along this
gradient, up to 900 m of altitude, abaxial leaf
stomata density decreases significantly compared
to the data from lowland sites (Table 2). In
mountainous conditions, short-wave solar
radiation often increases as well as the overall
incident sunlight beam. As a result, plant
transpiration increases with altitude as
demonstrated by Gale (1972). This is in
accordance with several authors who stated that
leaf stomata density increases from low- to upland
sites (Kofidis et al., 2007; Yang et al., 2007).
Especially, soil-water gradient was shown as the
main ecological factor discriminating leaf stomata
density along with elevation (Yang et al., 2007).
 Figure 3. SEM photographs showing different shape and position in the epidermis of stomata of the studied P. atlantica
populations, on the abaxial leaf side (x 1400). (A) Brezina, (B) Guerrara, (C) Berriane, (D) Elhamel, (E) Messaad, (F) Ain
oussara.
Our findings on the reduction of stomata density disagree
with the above-mentioned data suggesting that this leaf
trait is not solely driven by precipitations. Hence it is likely
that the edaphic environment would have played a key
role and this raises the question of the physiological
mechanisms involved in stomata density changes.
Because low soil-water retention capacity results in a
decrease of transpiration rate, we may argue that this
change might in turn impair the mobile flux of abscisic
acid and kinetin which are known to increase plant
stomata density (Wang et al., 1997; Franks and
Farquhar, 2001).
The stomata type recorded in this study were numerous.
From 764 m of altitude and for minimum temperature up
to 0°C, four stomatal types were recorded. However, for
minimum temperature below 0°C only two stomatal types
were shown. This is in accordance with the previous
studies (Kadi-Bennane et al., 2005; Smail-Saadoune,
2005) in which several types were recorded for P.
atlantica from three different populations whereas Lin et
al. (1984) reported only a single type (Table 3). In
addition, in our study, the laterocytic type was recorded;
this may be due to the adaptation of the species to the
harsh environmental conditions where it grows by
differentiating a new type of stomata. Plants occurring in
Mediterranean type climates are subjected to heat and
drought stress during the summer, most of them has
developed morphological and physiological mechanisms
which allow them to adapt and survive. These
mechanisms mainly comprise deeply developing stomata
and accumulation of secondary metabolites (Bosabalidis
and Kofidis, 2002). Drought is the environmental
condition that most affects leaf characteristics (XueJun
and XinShi, 2000). Drought stressed olive plants for
instance in order to save water, undergo anatomical
alterations in their leaves that, principally comprise an
increase of the stomatal density. With respect to stomatal
size, significant reductions of the length and width values
were estimated in the drought stressed plants as well as
increased stomata density on the lower leaf surface,
which contributes to a better control of transpiration
(Bosabalidis and Kofidis, 2002) whereas large stomata
seem to be a feature for plants living in humid conditions
(Hetherington and Woodward, 2003). In literature report,
with the exception of more advanced development of
palisade tissue, none of the characteristics commonly
associated with xeromorphic leaves were found in the
Pistacia species (Lin et al., 1984) which has been
referred to as xerophytes. We’ve showed in our previous
study (Belhadj et al., 2007a) that the covering hairs are
always associated with stomata, and located along the
midrib in the adaxial leaf face where stomata exclusively
occur. In addition to that, these stomata are always
covered with wax particles, and the epidermis was
strongly winkled, especially around the stomata.
CONCLUSION
Significant differences in stomata parameters among
populations were detected in this study. Stomata size on
the abaxial surface and stomata density on the adaxial
surface were the best discriminating traits. Oued Safene
(Os) site had the biggest stomata while Elhamel (E) site
had the smallest ones. The Messad (M) site registered
the highest stomatal density on the abaxial leaf surface
and the lowest on the adaxial surface one. However,
stomatal densities were the best characteristics related to
environmental conditions whereas their sizes were rather
a genotypic trait. These results along with our previous
observations on the morphological study and the
micromorphological as well as the palynological studies
(Belhadj et al., 2007a, b; Belhadj et al., 2008) provided
evidence that these variations may reflect phenotypic and
genotypic adaptations to native habitats. Such knowledge
might provide a basis for the selection of the appropriate
plant material for local reforestation programmes and/or
for the agronomical production, as this species is used as
rootstock for P. vera.
ACKNOWLEDGMENTS
We thank the laboratorio Regional de Sanidad Vegetal de
la Junta de Castilla la Mancha, for providing laboratory
facilities, the staff of the Electron microscope unit of the
Laboratoire des Mécanismes et des Transferts en
Géologie (LMTG) and the staff of the Ecolab UMR 5245
CNRS/UPS/INP, Toulouse, for their technical assistance
and co-operation. Also, Dr. Kevin J. Carpenter from the
University of California-Davis, U.S.A. and Dr. Odile
Décamps from the University of Toulouse, France for
their help in stomatal type identification.
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