doi:10.14720/aas.2020.115.2.1069 Original research article / izvirni znanstveni članek Introducing of some species of genus Allium subgenus Melanocrommyum from Iran as new sources of allicin Mahboobeh ZARE MEHRJERDI 1, 2, Mahdi MORIDI FARIMANI 3, Mahdi ABBAS MOHAMMADI 3, Jalal REZAEI 1 Received; March 02, 2019; accepted March 25, 2020. Delo je prispelo 02. marca 2019, sprejeto 25. marca 2020 Introducing of some species of genus Allium subgenus Mela- Uvajanje nekaterih vrst iz rodu Allium, podrodu Melanocrom- nocrommyum from Iran as new sources of allicin myum iz Irana kot novih virov alicina Abstract: Allicin is a sulfur compound found in genus Al- Izvleček: Alicin je žveplo vsebujoča snov v rodu Allium lium characterized by numerous biological and pharmacologi- s številnimi biološkimi in farmakološkimi lastnostmi. Podrod cal properties. Melanocrommyum, the second largest subgenus Melanocrommyum je drugi največji podrod v rodu Allium, v of Allium, has about 10 sections and 82 species in Iran. In this Iranu z okoli 10 sekcijami in 82 vrstami. V raziskavi so bili ana- study, allicin content of aerial part, aerial part fresh mass and lizirani nadzemni deli na vsebnost alicina in svežo maso pri 17 allicin yield belonging to 17 wild populations of six species of divjih populacijah šestih vrst iz rodu Allium ,sekcij. Acantho- Allium sect. Acanthoprason and Asteroprason growing in dif- prason in Asteroprason, ki rastejo na različnih območjih Irana. ferent region of Iran, were analyzed. Allicin content evaluation Vsebnost alicina, ovrednotena s HPLC metodo je pokazala using HPLC method showed its variation between populations razlike med populacijami v razponu od 26,98 do 58,11 mg from 26.98 to 58.11 mg g-1 FW, also showing that all the tested g-1 na svežo maso, kar kaže, da so vse populacije preiskušenih populations of Allium are rich in allicin. The average of aerial vrst bogate na alicinu. Poprečna sveža masa in vsebnost alicina part fresh mass and allicin yield varied between populations nadzemnih delov je med populacijami variirala od 0,49 g do from 0.49 g to 1.66 g and from 14 mg to 78 mg, respectively. 1,66 g in od 14 mg do 78 mg. Z generiranjem dendrograma po The populations were classified in four major groups using UPGMA metodi in klasterski analizi so bile populacije zdru- dendrogram generated by UPGMA method of cluster analysis. žene v 4 glavne skupne. Grupiranje populacij se ni popolnoma However, grouping of populations was not completely related ujemalo z vrstami in geografskimi regijami izvora. Ta raziskava to species and geographical regions. This study is the first evalu- je prva v Iranu, ki je ovrednotila vsebnost alicina v divjih po- ation of allicin content in wild populations of Allium sect. Acan- pulacijah vrst iz rodu Allium, Acanthoprason in Asteroprason. thoprason and Asteroprason in Iran. High amount of allicin in Zaradi velike vsebnosti alicina so vrste iz teh popuacij lahko these populations make them a new sources of allicin. njegov nov naravni vir. Key words: Acanthoprason; Asteroprason; medicinal Ključne besede: Acanthoprason; Asteroprason; zdravilne plant; population; variation; allicin content rastline; populacija; variabilnost; vsebnost alicina 1 University of Tehran, College of Aburaihan, Department of Horticulture, Tehran, Iran 2 Corresponding author, e-mail: [email protected] 3 Shahid Beheshti University, Medicinal Plants and Drug Research Institute, Department of Phytochemistry, Tehran, Iran Acta agriculturae Slovenica, 115/2, 341–347, Ljubljana 2020 M. ZARE MEHRJERDI et al. 1 INTRODUCTION knowledge about their potent in production of healthy metabolites is needed for domestication and breeding of Plants of the genus Allium belonging to mono- these species. cotyledonous flowering plants comprise more than 900 In this study allicin content of 17 wild populations accepted species, with main center of diversity in the belonging to six species of Allium sect. Acanthoprason eastern Mediterranean area, Southwest and Central Asia and Asteroprason which were collected from different re- (Fritsch & Abbasi, 2013). Allium species have been used gions of Iran, as a central part of diversity of this genus, for centuries as vegetables, as well as medicinal plants. were investigated. Several studies have been conducted showing Allium’s ther- apeutic properties as well as numerous reports referring to their antioxidant, antibacterial, antifungal, antiparasitic, 2 MATERIALS AND METHODS antiseptic, anti-inflammatory, anticancer, antidiabetic, cardioprotective, antiatherosclerosis, hepatoprotective 2.1 PLANT MATERIAL and immunomodulatory properties (Benkeblia, 2004; Galeone et al., 2006; Rizwani & Shareef, 2011; Feng et al., Seventeen populations of Allium including six spe- 2012; Lu et al., 2012; Nicastro et al., 2015; Sobolewska et cies of sect. Acanthoprason and Asteroprason from diverse al., 2015; Huang et al., 2016; Rad et al., 2017; Zeng et al., geographical origin which were described by Fritsch and 2017). Most of medicinal properties of Alliums are main- Abbasi (2013) were collected during the flowering stage. ly attributed to phenolic (like flavonoids: kaempferol, Table 1 gives the geographical location of populations. myricetin and quercetin derivatives) and sulfur-contain- All plants were cut 1cm above ground, weighted and kept ing compounds and beneficial elements such as selenium in a freezer at -80 oC. (Omar & Al-Wabel, 2010; Nwachukwu & Slusarenko, 2014; Soto et al., 2016). Organic sulphur compounds like alliin, allicin, allyl sulfide, (E)-ajoene, (Z)-ajoene and 2.2 SAMPLE PREPARATION 1,2-vinyldithiin are responsible for odor, flavor and most of biological activities of Alliums (Block, 1992; Benkeblia Allicin was extracted from randomly ten plants of & Lanzotti, 2007). Among these, allicin (diallylthiosulfi- each population in two replications according to Baghal- nate) has received more attention due to its significant ian et al. (2005). In brief, each 800 mg powder sample human health benefits (Oommen et al., 2004; Rahman, was placed in an ultrasonic bath at 4 oC for 5 min with 2007; Borlinghaus et al., 2014; Ye et al., 2016). 20 ml of distilled water. Tubes were incubated for 30 min Allicin’s structure and activities were described by at room temperature. The supernatant were then separat- Cavallito and Bailey in 1944 for the first time (Caval- ed by centrifuging at 6000 g for 30 min. The supernatant lito & Bailey, 1944). This unstable sulfur compound is (10 ml) were added to 15 ml of solution which contains composed from alliin by the action of alliinase released 1 % (v/v) solution of anhydrous formic acid and metha- from vacuoles upon crushing or damaging Allium tissues nol (4:6) and centrifuged at 6000 g for 5 min. The extracts (Jones et al., 2007). Allicin is now clearly accepted as a were analyzed as quickly as possible. biologically active compound, and several documents 20 mg butyl parahydroxybenzoate in 100 ml of have been published in this field (Ali et al., 2000; Li et methanol–water (50:50) was used as internal standard. al., 2010; Wallock-Richards et al., 2014; Gruhlke et al., 0.5 ml of internal standard was added to supernatant and 2017). Garlic (Allium sativum L.) is a main source of al- make up the volume to 10 ml and 20 μl of it was injected licin among cultivated Alliums. Nevertheless, there are into the HPLC. many wild Allium species that may have potentially some levels of allicin which needs investigation. Melanocrommyum, the second largest subgenus of 2.3 DETERMINATION OF ALLICIN Allium, comprises about 10 sections and 82 species in Iran (Fritsch & Abbasi, 2013). Some species in Acantho- The allicin were determined according to the meth- prason and Asteroprason, two sections in this subgenus, od of Baghalian et al. (2005). The HPLC analysis was car- are used by folk peoples as wild leafy vegetable and me- ried out on a Knauer HPLC system (Berlin, Germany) dicinal herbs. These plants have specific smell like garlic. equipped with a Knauer C18 column (25 cm × 4.6 mm) In spite of long local traditional usages, there is no re- and a PDA detector. The mobile phase was methanol– search on their beneficial compounds like allicin. These water (50 : 50) at a flow rate of 0.7 ml min-1. Elution was species are threatened with extinction because of wild- monitored at 254 nm. The percentage of allicin was cal- harvesting as the only way to reach them. Awareness and culated by using the following equation: 342 Acta agriculturae Slovenica, 115/2 – 2020 Introducing of some species of genus Allium subgenus Melanocrommyum from Iran as new sources of allicin Allicin (%) = "s1 m2 * 22.75" /"s2 m1" 3 RESULTS AND DISCUSSION Where s1 and s2 are the area of the peak corre- The aerial part allicin content of 17 populations of sponding to allicin and internal standard and m1 and Allium belonging to Acanthoprason and Asteroprason sec- m2 are the mass of the Allium powder and butyl par- tions collected from different regions of Iran are shown ahydroxybenzoate in internal standard solution, re- in Figure 1. The percentage of allicin content varied from spectively. The allicin content was expressed as mg -1g 26.98 to 58.11 mg g-1 FM, where the highest content was FM. found for Shen Jari population of A. pseudobodeanum, Mean allicin yield of each population (per plant) followed by Dehdasht of A. minutiflorum (57.95 mg g-1 were calculated by using percentage of allicin and aer- FM), Pir Baba Ali of A. subakaka (56.87 mg g-1 FM) and ial part fresh mass. Shirpala, another population of A. pseudobodeanum (55.94 mg g-1 FM), while the lowest content belonged to Taze Abad Oryeh population of A. kurdistanicum, fol- 2.4 DATA ANALYSIS lowed by Vali Abad of A. derderianum (27.42 mg g-1 FM). Variation in allicin content of different ecotypes of Pearson correlation and cluster analyses (UPG- garlic as a main source of this valuable metabolite is re- MA) were carried out on the data of allicin content and ported in previous studies (Baghalian et al., 2005; Wang aerial part fresh weight using the statistical software et al., 2014; Mostafa et al., 2015; Panahandeh et al., 2016).