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South African Journal of Botany 2003, 69(3): 422–427
Printed in South Africa — All rights reserved
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SOUTH AFRICAN JOURNAL
OF BOTANY
ISSN 0254–6299
Glandular trichomes and essential oils of Salvia glutinosa L.
A Kaya*, B Demirci and KHC BaÕer
Anadolu University, Faculty of Pharmacy, Pharmaceutical Botany, 26470 Eskisehir, Turkey
* Corresponding author, e-mail: aykaya@anadolu.edu.tr
Received 4 February 2003, accepted in revised form 29 April 2003
The aerial organs of Salvia glutinosa L. bear indumentum
with two types of trichomes: simple and multicellular
nonglandular trichomes, and stalked and sessile dense
glandular trichomes. Glandular trichomes are extremely
long-stalked and dense on the stem and calyx surfaces.
However, sessile glands are rare on the stem, calyx and
leaf adaxial surfaces and dense on the leaf abaxial
surface. Secretion accumulates in a subcuticular space
and is released to the outside by cuticle rupture. Water
distilled essential oil from dried aerial parts of S.
glutinosa was analysed by GC/MS. The main constituent
was identified as 1-octadecanol (11.6%).
The essential oil composition of aromatic plants of the family
Lamiaceae has been widely studied. The natural essential
oils have great commercial value. Little information is
available on the morphology, anatomy and development of
the glandular structures (trichomes) responsible for
secretion of these essential oils (Bosabalidis 1990, Maleci
and Servettaz 1991, Maleci et al. 1992, Servettaz et al.
1992, Özdemir and Ôenel 1999, Turner et al. 2000).
The genus Salvia L. with over 900 species is probably the
largest member of the family Lamiaceae and is found in both
subtropical and temperate parts of the world (Polunin and
Huxley 1967). Salvia is represented by 86 species in Turkey
(Hedge 1982). Since the most recent revision of the genus,
three new species (Davis et al. 1988, Vural and Ad2güzel
1996, Dönmez 2001) have been described; the total has
now reached 89. Many Salvia species are aromatic, rich in
essential oils and of potential economic interest besides
their ornamental uses. Many of these species are used to
flavour food as well as in cosmetics, perfumes and
pharmaceutical industries (Marin et al. 1996).
Salvia glutinosa L. is an aromatic plant that grows in
moist places in deciduous forest and scrub and in Picea
forests of north and south Anatolia and the flowering time
is from July to October (Hedge 1982). Its aerial organs
bear numerous glandular and non-glandular trichomes on
their surfaces. The composition of the oil of plants growing
in forests in Yugoslavia (Ivanic and Savin 1976) and
southern Italy (Senatore et al. 1997) has previously been
examined.
Previously, we reported the essential oil composition of
several Salvia species (BaÕer et al. 1993, 1995, 1996, 1997,
1998, Demirci et al. in press, Tümen et al. 1998). In
continuation of our studies on essential oil bearing plants,
this paper reports on the morphology and distribution of the
glandular trichomes of S. glutinosa and the chemical
composition of its essential oil.
Material and Methods
Plant material
Salvia glutinosa L. plants were collected during the flowering
period (August 2001) from ¤zmit (Keltepe) province of
Turkey. Voucher specimens were deposited in the
Herbarium of the Faculty of Pharmacy of Anadolu University
in Eskisehir, Turkey (ESSE 13943).
Scanning electron microscopy (SEM)
Leaves, stems and calyces were fixed with 3%
glutaraldehyde in 0.1M sodium phosphate buffer, pH 7.2 for
4h at 4°C. After washing the material was dehydrated by
acetone critical point drying. The specimens were mounted
on to SEM stubs using double-sided adhesive tape and
coated with gold. Photographs were taken with electron
microscope (Cam Scan S4).
Light microscopy
Transverse sections and surface prepations of leaves stems
and calyces were prepared manually for anatomical figures
of glandular trichomes and examined with a Leitz SM-LUX
binocular microscope with drawing tube.
Gas chromatography Mass spectrometry (GC MS)
The essential oil was analysed using a Hewlett-Packard
G1800A GCD system. Innowax FSC column (60m x
Introduction
South African Journal of Botany 2003, 69: 422–427 423
0.25mm Ø, with 0.25µm film thickness). Helium (1ml min–1)
was used as carrier gas. GC oven temperature was kept at
60°C for 10min and programmed to 220°C at a rate of 4°C
min–1 and then kept constant at 220°C for 10min then raised
to 240°C at a rate of 1°C min–1. Mass range was recorded
from m/z 35–425. Split ratio was adjusted at 50:1. Injection
port temperature was at 250°C. MS were recorded at 70eV.
Relative percentage amounts of the separated compounds
were calculated automatically from peak areas of the total
ion chromatograms. n-Alkanes were used as reference
points in the calculation of relative retention indices (RRI). A
library search was carried out using ‘Wiley GC/MS Library’
and ‘TBAM Library of Essential Oil Constituents’.
Results
Types of trichomes observed (Figures 1–7)
A. Heads unicellular A1: Short stalked (one basal epider-
mal cell and 1–3 stalk cells)
A2: Long stalked (one basal epider-
mal cell and 4–7 stalk cells)
B. Heads bicellular B1: Short stalked (one basal epider-
mal and one stalk cell)
B2: Long stalked (one basal epider-
mal cell and 1–2 stalk cells)
C. Peltate hairs: frequently short stalked (one basal epider-
mal cell, one stalk cell and four secretory cells or even
more)
D. Non-glandular multicellular trichomes, unbranched, unis-
eriate with cuticular micropapillae
Morphology and distribution of the glandular trichomes
The stems of S. glutinosa may rise to c. 1m. They are erect
and branched above and are ± densely glandular villous
above and hairy below. They have all types of trichomes
(A–D) (Table 1). The glandular trichomes are more variable
and the A2 type is more frequent on the stem (Figures 1, 2
and 3).
The leaves are simple, ovate-triangular, 8–14cm x
5–11cm, sagittate-hastate, serrate. Leaves of S. glutinosa
bear glandular (A1, C) and non-glandular trichomes (D) on
each site. Short capitate hairs (A1) which are composed of
one basal epidermal cell and one stalk cell are more fre-
quent than those with two stalk cells. C hairs are also easily
distinguished under the stereoscope. Their heads are four or
more celled and secrete essential oil which is formed at the
tip of the head between the raised cuticle and the apical cell
walls. Since the larger subcuticular space is filled with an
apparently foamy secretion (Figures 5, 7), the upper number
of the head cell is not determined. Non-glandular trichomes
(D) are found mainly on the ribs on the abaxial surfaces. Cell
number is up to five on the adaxial surfaces and up to seven
on the abaxial surfaces. A2 and B hairs are lacking com-
pletely on leaves (Figures 1, 4 and 5).
The calyx is tubular to campanulate, c. 12–17mm in fruit,
densely glandular-villous, upper lip 1 dentate, ± straight. The
distribution of trichomes on the calyces is particularly impor-
tant, since the calyx characters are often essential in the
taxonomic determination of Lamiaceae. The distribution of
trichomes on the outer calyces shows the same kind of hairs
as the stems (Table 1). A2 type trichomes are the most com-
mon. Small D type trichomes are abundant on the inner sur-
face while the glandular trichomes are absent (Figures 1, 6
and 7).
Trichome distribution on different plant parts in Salvia
glutinosa
Trichome distribution on the leaves (adaxial and abaxial sur-
faces), stem and calyx (inner and outer face) of S. glutinosa
is shown in Table 1.
Discussion
S. glutinosa bears numerous glandular and non-glandular
trichomes. Light (Figure 1) and scanning electron
microscopy (Figures 2–7) show details of the outer
Table 1: Trichome distribution on different plant parts in Salvia glutinosa. Symbols indicate: – = absence of hairs; ± = few hairs; +, ++ and
+++ = increasing presence of hairs.
Leaves Calyx
Hair type Adaxial Abaxial Stem Inner face Outer face
A1 + ++ ++ – ++
A2 – – +++ – +++
B1 – – ++ – ++
B2 – – + – ++
C ++ +++ ++ – ++
D++++
1++ +++ +
++1mainly on the ribs
A. Heads unicellular A1: Short stalked (one basal epidermal cell and 1–3 stalk cell)
A2: Long stalked (one basal epidermal cell and 4–7 stalk cell)
B. Heads bicellular B1: Short stalked (one basal epidermal and stalk cell)
B2: Long stalked (one basal epidermal cell and 1–2 stalk cell)
C. Peltate hairs: frequently short stalked (one basal epidermal cell, one stalk cell and four secretory cells or even more)
D. Non-glandular multicellular trichomes, unbranched, uniseriate with cuticular micropapillae
Kaya, Demirci and BaÕer
424
Figure 1: Glandular and non-glandular trichomes of S. glutinosa as viewed under a light microscope
Figures 2–7: Glandular and non-glandular trichomes of S. glutinosa in SEM. 2–3: A1, A2, B2, C and D trichomes on the stem. 4–5: C and D
trichomes on the abaxial surface of the leaf. 6–7: Numerous A2 and C trichomes on the outer surface of the calyx. Scale bars: 2 and 3 =
~200µm, 4 = ~100µm, 5 and 7 = ~50µm, 6 = ~500µm
South African Journal of Botany 2003, 69: 422–427 425
Kaya, Demirci and BaÕer
426
morphology of these trichomes. Glandular trichomes of S.
glutinosa are found in five different forms, i.e. A (A1, A2), B
(B1, B2) and C. Cross sections show the anatomy of the
glandular trichomes. In such sections, the capitate glandular
hairs consist of three cells (a unicellular foot, a unicellular or
multicellular stalk and a unicellular or bicellular head). The
peltate hairs were similarly observed to be composed of a
unicellular foot and a short stalk. Their heads, however, are
multicellular (four cells or even more). This pattern of
anatomy of the peltate hairs of S. glutinosa appears to be
similar to that described by Maleci and Servettaz (1991) and
Maleci et al. (1991, 1992).
On the basis of external morphology the glandular
trichomes of S. glutinosa are similar to those previously
described in Lamiaceae, namely for the leaves of Satureja
thymbra L. (Bosabalidis 1990) and Italian species of
Teucrium sect. Chamaedrys (Maleci and Servettaz 1991),
Teucrium marum L., Teucrium subspinosum Pourret ex Willd
(Servettaz et al. 1992) and Teucrium massiliense L. (Maleci
et al. 1992) and Salvia sclarea L. (Özdemir and Ôenel 1999).
In this study, besides the micromorphological
observations, we also report on the analysis of the volatile
compounds of S. glutinosa. The composition of the
essential oils from the aerial parts of S. glutinosa is reported
in Table 2. Dried aerial parts gave an essential oil yield of
0.1%. 1-Octadecanol (11.6%) was found as the major
constituent in the oil.
The essential oil composition of our material was found to
be quite different from those already reported. Bornyl
acetate was the main component in a sample from
Yugoslavia (Ivanic and Savin 1976) while γ-muurolene was
found as the major component for the leaves and flowering
tops of S. glutinosa growing in Italy (Senatore et al. 1997).
Also the absence of γ-muurolene could be a significant
feature of this wild growing species from Turkey while bornyl
acetate is represented with 3.2% in our sample.
Various factors, both endogenous and exogenous, can
affect the composition of the essential oil of S. glutinosa. We
believe that the time of flowering, geographical and climatic
factors may be very important. Several papers have
reported on the variation in the essential oil composition
induced by environmental, physiological and edaphic factors
which can induce changes in biosynthesis accumulation or
metabolism of given compounds of the essential oil
(Senatore et al. 1997).
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Table 2: The composition of the essential oil of Salvia glutinosa
RRI Compound %
1244 Amyl furan (2-Pentyl furan)tr
1304 1-Octen-3-one tr
1348 6-Methyl-5-hepten-2-one tr
1360 Hexanol 0.1
1391 (Z)-3-Hexenol tr
1393 3-Octanol 0.4
1400 Nonanal 2.1
1438 Hexyl 2-methyl butyrate 0.3
1452 1-Octen-3-ol 0.7
1466 α-Cubebene tr
1482 (Z)-3-Hexenyl-2-methyl butyrate 0.2
1496 2-Ethyl hexanol 0.1
1497 α-Copaene 2.0
1506 Decanal 0.6
1516 (E)-Theaspirane 1.6
1528 α-Bourbonene 0.1
1535 β-Bourbonene 2.9
1548 (E)-2-Nonenal 0.1
1553 Linalool 3.0
1553 (Z)-Theaspirane 1.2
1571 trans-p-Menth-2-en-1-ol 0.2
1590 Bornyl acetate 3.2
1600 β-Elemene 2.3
1612 β-Caryophyllene 4.6
1638 β-Cyclocitral 0.4
1650 γ-Elemene 0.4
1661 Alloaromadendrene 0.1
1663 Phenylacetaldehyde 0.1
1687 α-Humulene 3.5
1693 β-Acoradiene 0.6
1706 α-Terpineol 0.6
1719 Borneol 1.2
1726 Germacrene D 2.1
1741 β-Bisabolene tr
1758 cis-Piperitol 0.7
1766 Decanol 0.1
1773 δ-Cadinene 0.9
1815 2-Tridecanone tr
1827 (E,E)-2,4-Decadienal 0.6
1838 (E)-β-Damascenone 1.4
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2046 Norbourbonone 0.9
2050 (E)-Nerolidol tr
2071 Humulene epoxide-II 3.7
2131 Hexahydrofarnesyl acetone 3.1
2144 Spathulenol 2.0
2179 3,4-Dimethyl-5-pentylidene-2(5H)-furanone 1.7
2214 Phenyl ethyl tiglate 2.3
2300 Tricosane tr
2384 Hexadecanol tr
2384 Farnesyl acetone 0.2
2400 Tetracosane 0.1
2500 Pentacosane 0.2
2600 Hexacosane 0.1
2607 1-Octadecanol 11.6
2622 Phytol 1.3
2655 Benzyl benzoate 0.1
2740 Anthracene 0.1
2900 Nonacosane tr
Total 88.7
RRI = Relative retention indices calculated against n-alkanes
tr = Trace (< 0.1 %)
% calculated from TIC data
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Edited by J van Staden
South African Journal of Botany 2003, 69: 422–427 427
