Genetic, chemical and agro-morphological evaluation of the medicinal plant Origanum vulgare L. for marker assisted improvement of pharmaceutical quality [Elektronische Ressource] / submitted by Ali Azizi

Justus Liebig University Giessen
Institute of Crop Science and Plant Breeding I
Head: Prof. Dr. B. Honermeier



Genetic, chemical and agro-morphological evaluation of the medicinal plant
Origanum vulgare L. for marker assisted improvement
of pharmaceutical quality




Dissertation
Submitted for the degree of Doctor of Agricultural Science (Dr. agr.) to the Faculty of
Agricultural Sciences, Nutritional and Environmental Management
Justus Liebig University Giessen








Submitted by

Ali Azizi



Giessen 2010







This thesis was accepted as a doctoral dissertation in fulfillment of the requirements for the
degree of Doctor of Agricultural Science by Faculty of Agricultural Sciences, Nutritional
Sciences and Environmental Management, Justus-Liebig-University Giessen






Date of defence: July 22, 2010






The examination committee:

Chairman: Prof. Dr. Steffen Hoy
1. Supervisor: Prof. Dr. Bernd Honermeier
2. Supervisor: Prof. Dr. Dr. h.c. Wolfgang Friedt
Examiner: Prof. Dr. Sylvia Schnell
Examiner: Prof. Dr. Diedrich Steffens






























Meiner Frau Shiva
in Liebe
gewidmet




















DANKSAGUNG

Meinem Doktorvater, Herrn Professor Dr. Bernd Honermeier, danke ich herzlich für die
Überlassung des Themas und die engagierte Betreuung der Arbeit. Herrn Professor Dr. Dr. h.c.
Wolfgang Friedt danke ich als Zweitbetreuer für die hilfreiche Unterstützung bei fachlichen
Fragestellungen.

Herrn PD Dr. Feng Yan, Herrn Dr. Javad Hadian, Frau Dr. Carola Wagner und Herrn Dr. Michael
Gaudchau, danke ich für die freundliche Unterstützung bei der Interpretation der Ergebnisse, den
fachlichen Austausch und die wertvollen Diskussionen.

Herzlichen Dank an Edwin Mandler, Markus Kolmer, Pia Doernfeld, Rosa Allerdings, Stavros
Tzigos und Liane Renno sowie allen anderen Mitarbeitern des Instituts für Pflanzenbau und
Pflanzenzüchtung I für die technische Assistenz und Hilfe.

Besonders danke ich meiner Frau Shiva Torabi für ihre liebe Unterstützung und ihr stetes
Interesse am Fortgang der Arbeit.

Dem Deutschen Akademischen Austauschdienst (DAAD) gilt mein Dank für die finanzielle
Unterstützung der Arbeit über vier Jahre.

Ali Azizi



















TABLE OF CONTENTS


I. INTRODUCTION ................................................................................................................... 1

1.1. Origanum vulgare L. as a medicinal and spice plant .................................................... 1
1.2. Botany ........................................................................................................................... 1
1.3. Oregano essential oil ..................................................................................................... 2
1.4. Pharmacology ............................................................................................................... 3
1.5. Cultivation of Origanum vulgare L. ............................................................................. 4
1.6. Morphological, phytochemical and genetic diversity ................................................... 5
1.7. Breeding of oregano: conventional and biotechnological approaches........................... 6
1.8. Marker Assisted Selection (MAS .................................................................................. 7
1.9. Objectives ..................................................................................................................... 7


II. ARTICLES .............................................................................................................................. 9


Azizi A., Wagner C., Honermeier B., Friedt W.
Intraspecific diversity and relationships among subspecies of Origanum vulgare L.
revealed by comparative AFLP and SAMPL marker analysis.
Plant Systematic and Evolution (2009) 281: 151–160 .......................................................... 10

Azizi A., Yan F., Honermeier B.
Herbage yield, essential oil content and composition of three oregano
(Origanum vulgare L.) populations as affected by soil moisture regimes
and nitrogen supply.
Industrial crops and products (2009) 29: 554–561 ........................................................... 20

Azizi A., Hadian J, Honermeier B., Friedt W.
Associations between molecular markers, agro-morphological traits and
chemical characteristics in a germplasm collection of the medicinal
plant Origanum vulgare L. (Manuscript) .......................................................................... 28


III. DISCUSSION ...................................................................................................................... 57

3.1. Usefulness of AFLP and SAMPL markers for genetic studies of O. vulgare L .......... 57
3.2. Genetic, chemical and agro-morphological variations in O. vulgare L. ..................... 59
3.3. Enabling morphology and marker-assisted selection of O. vulgare L. ....................... 61 3.4. Effect of drought stress on oregano essential oil during the flowering phase ............ 63

IV. SUMMARY ....................................................................................................................... 65
V. ZUSAMMENFASSUNG ............................................................................................67
VI. REFERENCES ................................................................................................................ 69 I. INTRODUCTION


1.1. Origanum vulgare L. (oregano) as a medicinal and spice plant
Oregano is the common name for a general aroma and flavour primarily derived from more than 60
plant species used all over the world as a spice. Four main groups of plants commonly used for
culinary purposes can be distinguished, i.e., Greek oregano (Origanum vulgare ssp. hirtum (Link)
Ietswaart); Spanish oregano (Coridohymus capitatus (L.) Hoffmanns & Link); Turkish oregano
(Origanum onites L.); and Mexican oregano (Lippia graveolens HBK) (Lawrence, 1984).
In Europe and, in general, all over the world, the most commonly found oregano species belong to the
botanical genus Origanum so the commercial products of O. vulgare L. and O. majorana L. leaves are
known as oregano and marjoram on the market (Olivier, 1997). Because of special compositions of
essential oil, the leaves of Origanum plants are widely used as a very popular spice for food
production. European oregano (O. vulgare) is used as flavour in meat and sausage products, salads,
stews, sauces, and soups. Prior to the introduction of hops, oregano was used to flavour ale and beer
(Kintzios 2002a).
Recently, this spice plant has drawn more attention of consumers due to the antimicrobial, antifungal,
insecticidal and antioxidative effects of this herb on human healthy (Kokkini 1997, Kulisic et al. 2004,
Bakkali et al. 2008). As a medicinal plant, European oregano has traditionally been used as a
carminative, diaphoretic, expectorant, emmenagogue, stimulant, stomachic, and tonic. In addition, it
has been used as a folk remedy against colic, coughs, headaches, nervousness, toothaches, and
irregular menstrual cycles (Kintzios 2002a).

1.2. Botany
The genus Origanum belongs to the family Lamiaceae and comprises 43 species and 18 hybrids
widely distributed in Eurasia and North Africa (Ietswaart 1980, Duman et al. 1998). The species
Origanum vulgare L. is predominate in occurrence and the most variable species of the genus and the
only one commonly known as `oregano' in the most European countries (Vokou et al. 1993, Tucker
and Maciarello 1994).
Origanum vulgare L. is a beautiful plant, flowering in heady corymbs, with reddish bracts and purple
corollas. The plant flowers in late summer, grows in spikes, and is purplish white. The plant height is
30–60 cm with comparable width (Kokkini 1997). One of the considerable morphological
characteristics of the Origanum plants is the presence of glandular and nonglandular hairs (peltate
hairs or glandular scales) covering the aerial organs. Both types of hairs originate from epidermal cells
(Netolitzky 1932). The glandular hairs are numerous on the vegetative organs such as stems, leaves
and bracts, while their density becomes reduced on the reproductive organs such as calyces and
corollas (Bosabalidis and Tsekos 1984). The glandular hairs produce and secrete an essential oil wi