Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 81–88 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal Research article COMPARISON OF COLCHICINE CONTENT BETWEEN HYSTERANTHOUS AND SYNANTHOUS COLCHICUM SPECIES IN DIFFERENT SEASONS Alirezaie Noghondar Morteza1*, Arouee Hossein 2, Shoor Mahmoud 2, and Rezazadeh Shamsali 3 1*PhD student, Ferdowsi University of Mashhad, Agriculture Faculty, Horticultural Sciences Department, Mashhad, Iran 2 Assistant Professor, Ferdowsi University of Mashhad, Agriculture Faculty, Horticultural Sciences Department, Mashhad, Iran 3 Assistant Professor, Institute of Medicinal Plants, Department of Pharmacognosy and Pharmaceutics, ACECR, Tehran, Iran *Corresponding author: Email: [email protected] Received: 13/12/2012; Revised: 24/01/2013; Accepted: 30/01/2013 ABSTRACT In order to compare of different phonological stages and seasonal changes of colchicine content between hysteranthous and synanthous colchicum species, amount of colchicine was determined in Colchicum speciosum Steven, C. kotschyi Bioss and C. robustum Stefanov, in different seasons, 2009–2010. The observations under wild conditions showed, that the leaves of appeared with flowers in the same stage of life cycle (synanthous) in C. robustum, while in case of C. kotschyi and C. speciosum flowers occurred first and leaves later, in another developmental stage (hysteranthous). Seed’s colchicine content in C. robustum, C. kotschyi and C. speciosum was obtained as 1.28, 0.46 and 0.92 mg g-1 dry weight, respectively. Corm’s colchicine content was higher in C. speciosum than the other species in all seasons. The highest colchicine content of corm in C. speciosum was obtained in winter and autumn (2.17 and 2.13 mg g-1 dry weight, respectively), while in C. robustum and C. kotschyi it was found in autumn, 0.49 and 0.77 mg g-1 dry weight, respectively. The lowest colchicine content of corms was obtained in summer, when the corms were dormant before flowering stage, in C. speciosum and C. kotschyi, 0.131 and 0.0058 mg g-1 dry weight, respectively, whilst in C. robustum obtained in winter, 0.08 mg g-1 dry weight, synchronous to flowering and vegetative growth. KEYWORDS: Colchicine content, Colchicum kotschyi, C. speciosum, C. robustum, Flowering stage, Hysteranthous, Root activity, Synanthous, Seasonal changes. Cite this article: Alirezaie Noghondar Morteza, Arouee Hossein, Shoor Mahmoud, and Rezazadeh Shamsali (2013), COMPARISON OF COLCHICINE CONTENT BETWEEN HYSTERANTHOUS AND SYNANTHOUS COLCHICUM SPECIES IN DIFFERENT SEASONS., Global J Res. Med. Plants & Indigen. Med., Volume 2(2): 81–88 Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI || Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 81–88 INTRODUCTION cilicicum were 0.05%, 0.01%, 0.01% and 0.20% (g% dry weight), respectively (Sütlüpinar et al., The genus Colchicum belongs to the family 1988). In another study by Alali et al. (2004), C. Colchicaceae, which comprises of 19 genera, stevenii corms, flowers and leaves were reported and 225 species (Nordenstam, 1998). Plants of to contain 0.17, 0.12 and 0.20 (wt/wt) g%, the genus Colchicum have been known for more respectively, while C. hierosolymitanum corms than 2000 years for their marked beneficial and and flowers were found to contain 0.13 and 0.09 poisonous effects (Brickell, 1984). The modern (wt/wt) g%, respectively. Ondra et al. (1995), medicine uses Colchicum as a source of assayed corms of seven Turkish Colchicum therapeutically active alkaloids called species; namely: C. macrophyllum, C. turcicum, colchicinoids. One of the most abundant C. cilicicum, C. kotschyi, C. bornmuelleri, C. alkaloid - colchicine, is known to have speciosum and C. triphyllum for their cancerostatic, antirheumatic, antimitotic, colchicinoid alkaloids. Colchicine content was antiinflammatory, cathartic and emetic effects. It found to be 222.3, 323, 300, 1058, 3063, 4245 is also applied in plant breeding to induce and 958 µg g-1 dried drug, respectively. polyploids (Komjatayova et al., 2000; Frankova et al., 2005). In addition to the genus Colchicine variation in different organs of Colchicum, colchicine was reported from plant and during different growth stages has species belongs to Merendera and Gloriosa been studied by researchers. Colchicine and genera, which belonging to the Colchicaceae demecolcine were determined in raw and dried family (Nordenstam, 1998). leaves, stems, mother and daughter corms of C. autumnale in four stages of its ontogenesis by Very little is known about the factors Vicar et al. (1993). They found that colchicine interfering with the biosynthesis of colchicine- content in raw material varies during plant like alkaloids. Results obtained by Sütlüpinar et growth. Colchicine content in C. brachyphyllum al. (1988), indicated that the composition of and C. tunicatum, was determined during tropolone alkaloids differs in different parts of different growth stages by Alali et al. (2006). the plants and varies during the different growth Underground parts in both species and during stages (Sütlüpinar et al., 1988). Presence and different growth stages, always showed higher concentration of colchicine is determined by a colchicine content than the above ground parts. variety of environmental factors including In C. brachyphyllum, total colchicine content of season (Vicar et al., 1993; Poutaraud and underground parts during flowering stage was Girardin 2002; Alali et al., 2006) and resource found to be about 0.15% (wt/wt), while that of availability (Hayashi et al., 1988; Pouraraud and aerial parts was only about 0.04% (wt/wt). In C. & Girardin, 2005; Mróz, 2008) as well as tunicatum, total colchicine content of genetic variations between populations and underground parts was found to be 0.12% individuals (Poutaraud and Champay, 1995). (wt/wt) and 0.13% (wt/wt) during flowering and Also colchicine content varies among different vegetating stages, respectively, while that of organs of the plant body (Sütlüpinar et al., 1988; aerial parts was only about 0.04% (wt/wt) and Alali et al., 2004; Alali et al., 2006). 0.02% (wt/wt), respectively (Alali et al., 2006). Among all species of Colchicum, C. Generally, geophytes are plants that survive autumnale is the best source for colchicine. The by subterranean storage organ with renewal richest plant parts in colchicine are the corms buds (Raunkiaer, 1934). They divide into two and seeds. C. autumnale seeds contain 0.6-1.2%, groups – synanthous and hysteranthous one. The while corms contain up to about 0.6%. Seeds are leaves of synanthous geophytes coexist with mainly used by the pharmaceutical industry for flowers in the same stage of life cycle. In case of the extraction of colchicinoids (Trease & Evans, hysteranthous plants flowers occur in the first 1983). The content of colchicine alkaloid in and leaves later, in another developmental stage corms, stems, leaves, and flowers of C. (Dafni et al., 1981). A special case is the Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI || Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 81–88 hysteranthous plant Colchicum tunicatum which during 2009–2010. Observations were including perceive the photoperiodic signal when the dry of developmental stages such as beginning of bulb lies well below the soil surface (Halevy, flowering, peak flowering time, root formation 1990). C. speciosum, C. kotschyi Boiss, and C. time, beginning of vegetative growth, fruiting robustum stefanov, are three wild growing and capsule formation and daughter corm Iranian Colchicum species (Presson, 1992). C. formation in wild conditions. speciosum Steven and C. kotschyi Bioss are hysteranthous but C. robustum is a synanthous Extraction and Isolation species (Presson, 1992). The methods described by Rosso and So far no study has been performed on Zuccaro (1998) and Alali et al. (2006), were colchicine content variation between synanthous adopted with some modifications. Acetonitrile, and hysteranthous Colchicum species in methanol and other reagents were of different seasons, thus the aim of this study was chromatographic grade and prepared from to evaluate phenological changes and their Panreac (Spain). Reference standard of relationship with corm and seed colchicine colchicine was prepared from USP. content variation among three Iranian native Colchicum species, under their habitat The corms were sliced into small pieces and conditions. air-dried at room temperature together with the seeds. After drying, exact weight of 2 g of MATERIAL AND METHODS corms (collected in different seasons) and 2 g of seeds of three Colchicum species were grounded Plant Material to powder in a laboratory mill and then used for extraction. Powdered material placed into The corms of three wild Colchicum species 250 mL Erlenmeyer flasks and extracted with were collected in different seasons (spring, 100 mL of methanol in 35oC for 1h with summer, autumn and winter during 2009–2010, ultrasonic apparatus. Afterwards, plant residues and seeds were collected in spring 2010. Corms were filtered through Wattman filter paper and and seeds of C. speciosum, C. robustum and C. the filtrates were saved. Then plant residue was kotschyi were collected from Khalkhal-Asalem transferred into Erlenmeyer flasks again and road, Ardabil province, at an altitude of 940m in extracted with 50mL of methanol in 35oC for Iran, Babaaman