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Intraspecific variability of wild marjoram
(Origanum vulgare L.) naturally occurring in Poland
Z. W’glarz 1; E. Osidska; A. Geszprych; J. Przybyb
1
Department of Vegetable and Medicinal Plants, Warsaw Agricultural University, Nowoursynowska 166, 02-787
Warszawa, Poland
ABSTRACT: Intraspecific variability of wild marjoram (Origanum vulgare L.) naturally occurring in Poland.
Developmental and chemical analysis of six populations of wild growing Origanum vulgare L. was done. The most
segregating morphological and developmental traits were: plant habit, shape and size of leaves, colour of flowers
(white, pink, and dark pink) and dry mass of herb (from 243.1 to 636.6 g·plant-1). The content of essential oil in airdry herb collected at the full blooming stage ranged from 0.35 to 0.55%, and the yield of essential oil – from 1.4 to
3.2 ml·plant-1. The main constituents of essential oil were: sabinene, terpinene-4-ol, â-pinene and linalool. The
studied populations differed in respect of the content of phenolic acids and flavonoids. In all populations the dominant
phenolic acid was rosmarinic acid and the dominant flavonoid compound was luteolin 7-O-glucoside.
Key words: morphological traits, herb, essential oil, flavonoids, phenolic acids
INTRODUCTION
Origanum is one of the most variable genera
of Lamiaceae family. In the north part of Europe, from
England to Siberia, commonly found is Origanum
vulgare subsp. vulgare – wild marjoram (Ietswaart,
1980; Kokkini 1997). This species occurs naturally
also in Poland. Wild marjoram is a perennial, growing
on dry sunny slopes and forest margins in rather small
clusters. The herb of this plant is collected from natural
sites and used as a raw material in the
pharmaceutical, cosmetic and food industry. It is also
a commonly used seasoning (Melchior & Kastner,
1974, Tucker & Maciarello, 1994). The distribution of
this species on the area of Poland is irregular. Longterm observations indicate that its natural sites occur
mainly in the eastern Poland. In this region the highest
phenotype variability of wild marjoram was also found
(Osiñska & Pióro-Jabrucka, 2003).
The aim of the investigation was to determine
the developmental and chemical variability of six wild
marjoram populations originating from the middle-east
of Poland.
These studies are the part of the national
programme on biodiversity conservation and
sustainable exploitation of Polish wild growing
medicinal plants.
MATERIAL AND METHOD
The study was carried out in 2003 at the
Department of Vegetable and Medicinal Plants,
Warsaw Agricultural University. The experimental
Recebido para publicação em agosto/2004
Aceito para publicação em julho/2006
material were two-year-old plants of six populations
of wild marjoram (Origanum vulgare L.) The plants
were grown from the seeds collected in 2002 from
six natural sites located in the middle-east of Poland.
The experiment was established by the method of
randomised blocks with four replications on the plots
of 20 m2. The plant spacing was 50 x 60 cm. The
morphological and developmental observations were
done. The herb was collected at the full blooming
stage and dried at 35ºC.
The chemical analyses were done in four
replications – a replication was the sample of the
herb from one field replication.
The content of essential oil was determined
according to Polish Pharmacopoeia VI. The
identification of essential oil constituents was
performed by gas chromatography with the
parameters as follows: gas chromatograph HewlettPackard 6890; capillary column HP-5, length 30 m,
diameter 0.32 mm; detector temperature 250ºC;
injector temperature 220ºC; carrier gas – helium.
Column temperature was programmed as follows:
60ºC (2 min), temperature increment 8ºC/min, final
temperature 280ºC (5 min). The separated
compounds were identified on the basis of retention
times of available standards.
Sample for HPLC analysis was prepared as
follows: 1 g of dried raw material was extracted with
100 ml MeOH in modified Soxhlet apparatus (Büchi
B-811) and evaporated to dryness. The residue was
dissolved in 10 ml of MeOH and filtered (Iso-DiscTM
0.45 ìm). HPLC analysis was performed on Shimadzu
chromatograph with SPD-M10A VP DAD detector;
column: LiChrospher RP18 5 ìm 250 x 4,6 mm
(Supelco) at 26ºC; mobile phase: (A) 10% ACN and
(B) 55% ACN (Merck) with the flow rate 1 ml·min-1;
Rev. Bras. Pl. Med., Botucatu, v.8, n.esp., p.23-26, 2006.
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TABLE 1. Morphological characteristics of plants
TABLE 2. Yield of air-dry herb and essential oil
TABLE 3. Main volatile compounds (% in essential oil)
detection wave: 254 nm. Peak identification was
confirmed by comparison of retention time and
spectral data with adequate parameters of standards
(Sigma, Roth, Extrasynthese).
The results were subjected to one-way
analysis of variance. The significance of differences
was determined by the Tuckey’s test at the
significance level 0.05.
RESULT AND DISCUSSION
The most visible segregating morphological
traits of the investigated populations were: colour of
flowers (white, pink, and dark pink), habit of plants
(from vertical to semi-horizontal) and size and shape
of leaves (Table 1, Figure 1).
The populations significantly differed in
respect of the mass of herb. The highest yield of herb
per one plant was observed at the population A (636.6
g·plant-1), and the lowest at the populations C and F
(276.1 and 243.1 g·plant-1, respectively). The content
of essential oil in air-dry herb collected at the full
blooming stage ranged from 0.35% (population E) to
0.55% (population B). According to Arnold et al.
(2002), the content of essential oil in Origanum sp.
may come up even to 8.8%. The highest yield of
essential oil was obtained from the herb of population
A (3.2 ml·plant-1). It was almost three times higher
than that obtained from the population F (Table 2).
The presence of essential oil in wild
marjoram herb is specially important because of its
sensory value and application in food industry.
Numerous studies indicate high variability of the
composition of wild marjoram essential oil (Arnold et
al., 2000; Burdock, 2000; D’Antuono et al., 2000;
Melegari et al., 1995; Skoula et al., 1999).
In the essential oil from the herb of all
Rev. Bras. Pl. Med., Botucatu, v.8, n.esp., p.23-26, 2006.
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FIGURE 1. Shape of leaves
investigated populations relatively high percentage of
sabinene was determined. It was the dominant
compound in the essential oil of population E (Table
3). This compound was also reported by Nykänen
(1986) and D’Antuono et al. (2000), but in those
investigations its content in essential oil of wild
marjoram was rather low. In the population A high
content of tepinene-4-ol and linalool in essential oil
was observed. The wild marjoram plants characterised
by high content of linalool were found also in Spain
(Garcia & Sanz, 2001), and in north Italy (D’Antuono
et al., 2000). In the essential oil of population F the
main compound was â-pinene. This population was
also characterised by relatively high percentage of
thymol. In the essential oils of three populations (A,
B, C) â-caryophyllene and caryophyllene oxide were
present in considerable amount. One of the major
compounds in essential oil of population B, C, D, E
was cineol. Taking into consideration the results of
chromatographic analysis, it can be stated that the
investigated population differ in the composition of
essential oil and they seem to be also different from
wild marjoram originating from other regions of Europe.
In available literature there is no data on the
composition of phenolic compounds in wild marjoram
herb. In the herb of the investigated populations twelve
phenolic compounds were identified – six phenolic
acids and six flavonoids. In all populations the
dominant phenolic acid was rosmarinic acid. High
content of o-coumaric acid in herb of the investigated
populations was also found. The most distinct
differences between populations were observed in
respect of the content of rosmarinic, vanillic and ocoumaric acids. The dominant flavonoid compound
was luteolin 7-O-glucoside. The content of this
compound and rutin in the herb of investigated
populations was the most variable (Table 4).
CONCLUSION
1. Ex situ study on six wild growing populations of
Origanum vulgare L. indicated their
developmental and chemical variability.
2. The most segregating morphological and
developmental traits were: plant habit, shape
and size of leaves, colour of flowers and dry
mass of herb.
3. The populations differed in respect of the essential
oil content in herb and its composition.
4. The main constituents of essential oil in the herb
of the investigated populations were: sabinene,
terpinene-4-ol, â-pinene and linalool.
5. In all populations the dominant phenolic acid was
rosmarinic acid, and the dominant flavonoid
compound was luteolin 7-O-glucoside.
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