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Genetic and Chemical Diversity in Perovskia Abrotanoides KAR

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Genetic and Chemical Diversity in Perovskia Abrotanoides KAR (1 of 14) e1700508 Genetic and Chemical Diversity in Perovskia abrotanoides KAR. (Lamiaceae) Populations Based on ISSRs Markers and Essential Oils Profile Seyyed Hossein Pourhosseini,a Javad Hadian,a Ali Sonboli,b Samad Nejad Ebrahimi,c and Mohammad Hossein Mirjalili*a aMedicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Evin, 1983969411 Tehran, Iran, e-mail: [email protected] bDepartment of Biology, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Evin, 1983969411 Tehran, Iran cDepartment of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Evin, 1983969411 Tehran, Iran Genetic and the essential oil composition variability among twelve Perovskia abrotanoides populations (PAbPs) growing wild in Iran were assessed by ISSR markers, GC-FID and GC/MS, respectively. Nine selected ISSR primers produced 119 discernible bands, of them 96 (80.7%) being polymorphic. Genetic similarity values among populations ranged between 0.07 and 0.79 which indicated a high level of genetic variation. Polymorphic information content, resolving power and marker index generated by ISSR primers were, 0.31, 6.14, and 3.32, respectively. UPGMA grouped PAbPs into four main clusters. Altogether, 38 chemical compounds were identified in the oils, and a relatively high variation in their contents was found. Camphor (11.9 – 27.5%), 1,8-cineole (11.3 – 21.3%), a-bisabolol (0.0 – 13.1%), a-pinene (5.9 – 10.8%), and d-3-carene (0.1 – 10.5%) were the major compounds. Oxygenated monoterpenes (32.1 – 35.8%) and monoterpene hydrocarbons (25.7 – 30.4%) were the main groups of compounds in the oils studied. Cluster analysis and principal-component analysis were used to characterize the samples according to oil components. Four main chemotypes were found to be Chemotype I (camphor/1,8-cineol), Chemotype II (1,8-cineole/camphor), Chemotype III (camphor/1,8-cineol/a-bisabolol), and Chemotype IV (camphor/d-3-carene/ a-bisabolol). The information, provided here on P. abrotanoides populations, will be useful to introduce this plant into agricultural systems. Keywords: Perovskia abrotanoides, essential oils, genetic diversity, ISSR, chemotype. conditions persist in the plant’s habitat, next genera- Introduction tions are chosen to adapt to the new environment Medicinal and aromatic plants (MAPs) changed their and this adaptation gradually becomes inherited and chemical profile in different ecological conditions in can be transmitted to these generations. In the other order to adapt to the environment.[1][2] Therefore, the hand, wild populations of MAPs are heterogeneous in populations of a medicinal species that are growing in morphological and chemical characteristics.[7][8] In this various natural habitats show the variability in the case, if a medicinal species is to be introduced into quantity and quality of active ingredients, which lead agricultural systems because of its economic impor- to differences in their pharmacological and biological tance and in particular, the risk of the occurrence of activity.[3 – 5] The genetic flexibility of plant popula- such populations, the genetic structure as well as tions makes this variation possible and then gradually chemical diversity of its natural populations in order leads to arise some individuals which are different in to provide the raw materials with security, stability chemical and botanical characteristics.[6] When the and proper function should be firstly investigated. plant is exposed to environmental changes, variability Because of the climate diversity, Iran has a vast and occurs in its physiological and chemical behavior to unique biodiversity especially for MAPs. Flora of Iran is adapt to the new environmental conditions. These represented by 7500 plant species, of which 1700 are changes are usually unstable, but if the environmental MAPs.[9] Of course, any efforts to evaluate the DOI: 10.1002/cbdv.201700508 Chem. Biodiversity 2018, 15, e1700508 © 2018 Wiley-VHCA AG, Zurich, Switzerland Chem. Biodiversity 2018, 15, e1700508 phytochemical and genetic characteristics of each spe- population genetic,[39][40] and phylogenetic studies.[41] cies can lead to the introduction of susceptible spe- According to the literature information, ISSR markers cies for the production of new drugs. have been efficiently used for the genetic analysis of The Lamiaceae, with 46 genera and ca. 420 species several medicinal plant species.[42 – 44] and subspecies, have a great diversity and distribution Phytochemical and molecular markers have been in the flora of Iran.[10 – 12] Perovskia, a small genus simultaneously used to characterize the level of varia- from this family, is distributed in various regions of tion in several medicinal plant species, such as Witha- Asia, as Iran, Afghanistan, and Pakistan. The genus is nia spp.,[45] Ocimum basilicum,[46] Zataria multiflora,[47] represented in Iran by three species viz. Clitoria ternatea,[48] Salvia sclarea,[35] Ocimum selloi,[32] [49] [37][50] P. abrotanoides KAR., P. atriplicifolia BENTH., and Ocotea spp., and Satureja rechingeri. In the [13] P. artemisoides BOISS. Perovskia species are known to present study, ISSR markers were used for the first contain different class of compounds such as essential time to reveal the extent and distribution of the oils (EOs), phenolics, flavonones, irregular triterpenes, genetic diversity of twelve P. abrotanoides populations steroids and their glycoside and a large amount of (PAbPs) from Iran, as a first step towards gaining a abietane-type norditerpenoidquinones which called better knowledge of the genome diversity of the tanshinones.[14 – 18] P. abrotanoides, with the common plant. We also evaluated the EOs variability of the Persian name of ‘Brazambel’, is an aromatic erect herb wild-growing populations, which is important for phar- which is mainly growing in mountains from Northeast- maceutical and other related industries. ern Iran across Northern Pakistan to Northwestern India.[19] The plant is used by local communities for Results and Discussion treatment of leishmaniasis, typhoid, fever, headache, Habitats Characteristics of Perovskia abrotanoides gonorrhea, vomiting, motion, toothache, atherosclero- sis, cardiovascular diseases, liver fibrosis, and The geographic distribution of the studied cough.[20 – 23] It has sedative, analgesic, antiseptic and P. abrotanoides populations (PAbP1 – PAbP12) and cooling effect.[22][24] The plant herbal tea is used in P. atriplicfolia (PAtP) is presented in Table 1 and Fig- curing infection problems and painful urination.[25] ure 1. They belonged to the different geographical The essential oils of P. abrotanoides play an important zones. PAbP1 to PAbP12 and P. atriplicifolia population role in protection of stored grains and showed to be (PAtP) were located at the northeast and center of Iran effective in washing wounds, anti-ring worm, dermal with an inferior semi-arid climate characterized by a parasites, anti-fungus and anti-hypoxia.[26 – 28] The mean rainfall of 150 – 475 mm/year. The altitudes chemical composition of P. abrotanoides oil has been ranged from 1055 m (PAbP9) to 2220 m (PAbP2) previously investigated,[27 – 30] and 1,8-cineole, d-3- (Table 1). As can be shown in Figure 1, the studied carene, camphor, myrcene, and b-caryophyllene have Perovskia populations have grown within latitude of been reported as the major components of the oils. 33°350Nto37°250N and longitude of 50°590Eto Relationships among genetic and chemical variation 58°170E. have been studied at different levels in plants.[31 – 35] Species diversity in an environment depends on However, it should be noted that use of chemical pro- the ability to produce and sustain ecosystem.[51] file is most useful in taxonomic classification only Reducing biodiversity may be due to the environmen- when other complicating factors, such as environmen- tal effects and reducing the fertility of the plant com- tal conditions, plant developmental stages, and extrac- munity. There are many reports in the literature tion methods are taken into account.[2][34] regarding the variation in the chemical profile of EOs Molecular markers are very useful in an early from various plant species collected from different breeding program for allowing germplasm screening geographical regions.[47][52] Such differences could be at any developmental stage of the plants. Among dif- linked to the varied environmental factors and possi- ferent markers, DNA-fingerprinting techniques are ble adaption response of different populations, result- independent from environmental effects, unlimited in ing in different chemical products being formed, number, and show high level of polymorphism. without morphological differences being observed in Molecular markers provide a powerful tool for proper the plants.[53] Altitude, temperature, annual precipita- characterization of germplasm and chemotypes and tion as well as soil texture has been reported as the their management. Among developed genetic mark- major environmental factors which affected the chem- ers, inter-simple sequence repeat (ISSR) markers have ical composition of the EOs.[54] Therefore, in the case been widely used for plant diversity analyses based of exploitation and introduction of a medicinal species on DNA finger-printing,[36] genetic diversity,[37][38] into the mass cultivation system, the study of its www.cb.wiley.com (2 of 14) e1700508 © 2018 Wiley-VHCA AG, Zurich, Switzerland Chem. Biodiversity 2018, 15,
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