ORIGINAL ARTICLES Bioforsk -Norwegian Institute forAgricultural and Environmental Research1,Aas; Bioforsk –Norwegian Institute for Agricultural and Environmental Research, Bioforsk Øst Apelsvoll2,Kapp,Norway Bioactive compounds produced by clones of Rhodiola rosea maintained in the Norwegian germplasm collection A. Elameen 1,S.Dragland 2,S.S.Klemsdal 1 Received October 22, 2009,accepted January 15, 2010 Dr.Sonja Sletner Klemsdal, Bioforsk -Norwegian Institute for Agricultural and Environmental Research,Plant Health and Plant Protection Division, Department of Genetics and Biotechnology,Hogskoleveien 7, N-1432 Aas, Norway [email protected] Pharmazie 65: 618–623 (2010) doi: 10.1691/ph.2010.9806 Roseroot, Rhodiola rosea,isaperennial herbaceous plant of the family Crassulaceae.The rhizomes of 95 roseroot clones in the Norwegian germplasm collection were analysed and quantified fortheir content of the bioactivecompounds rosavin, salidroside,rosin, cinnamyl alcohol and tyrosol using HPLC analysis.All five bioactivecompounds were detected in all 95 roseroot clones butinhighly variable quantities.The ranges observedfor the different compounds were forrosavin 2.90–85.95 mg g−1,salidroside 0.03–12.85 mg g−1, rosin 0.08–4.75 mg g−1,tyrosol 0.04–2.15 mg g−1 and cinnamyl alcohol 0.02–1.18 mg g−1.The frequency distribution of the chemical content of each clone did not reflect acertain geographic region of origin or the gender of the plant. Significant correlations were found forthe contents of severalofthese bioactive compounds in individual roseroot clones.Alow,but not significant correlation wasfound between AFLP markers previously used to study the genetic diversity of the roseroot clones and their production of the chemical compounds.The maximum levelofrosavin, rosin and salidroside observedwere higher than forany roseroot plant previously reported in literature,and the roseroot clones characterized in this study might therefore prove to be of high pharmacological value. 1. Introduction Investigations on the phytochemistry of roseroot have revealed the presence of about 28 compounds classified into six distinct Roseroot, Rhodiola rosea,orarctic root also commonly known groups (Brown et al. 2002). The main biologically active com- as golden root, is aperennial herbaceous plant of the family pounds are salidroside, tyrosol, cinnamyl alcohol glycosides, Crassulaceae.Roseroot is distributed from China into the rosavin, rosarin and rosin (Kucinskaite et al. 2007; Wiedenfeld mountain regions of Central and Northern Europe, in the et al. 2007). There are nearly 200 Rhodiola species butcinnamyl coastal regions of North America as well as in Russia and alcohol glycosides and monoterpenoid, rosiridol and its gluco- in the FarEast. Historically this plant has been described side rosiridin have been found only in R. rosea (Ganzera et al. as an adaptogen (Kelly 2001), and Vikings used roseroot to 2001). It has previously been shown that the total flavonoid con- enhance their physical strength and endurance (Magnusson tent of the Nordic plants ranged from 0.34 to 0.51%, while those 1992). In Russia, Northern Europe, China, and more recently in of German and Austrian origin had acontent of 0.12–0.29% USA, roseroot has been used as atraditional herbal medicine, (Galambosi et al. 1999). valued for its ability to enhance human resistance to stress HPLC has been used to characterize and quantify the chemi- or fatigue (Sanderbergand Bohlin 1993; Darbinyan et al. cal composition in several plants including Rhodiola species. 2000). It promotes longevity and has been claimed to have Globally,more than 60% of the total pharmaceutical market antiallergenic, anti-depression and anti-inflammatory effects, to is derivedfrom plants (Wakdikar 2004). The composition and result in enhanced mental alertness and give cardio-protection, contents of chemical compounds are consequently one of the and has been used for avariety of therapeutic applications most important traits in acommercial breeding program of such including cancer therapy(Kurkin and Zapesochnaya 1986; plants and agrouping of accessions in corresponding germplasm Maslovetal. 1997; Razina et al. 2000; Spasovetal. 2000; De collections based on their chemical composition would be use- Bock et al. 2004; Mattioli and Perfumi 2007). ful. Clones with high chemical content may carry genes that The chemical composition of roseroot has been widely studied can be used to improve R. rosea collections. So farnatural wild (Dubichevetal. 1991; Kurkin et al. 1986; Furmanowa et al. resources of Rhodiola species have been harvested and used as 1999; Ming et al. 2005; Galambosi et al. 2007). The active rawmaterial for the pharmaceutical market. The natural growth metabolite salidroside and its aglycon tyrosol (Kuryanovetal. of Rhodiola species is mostly limited to quite specific areas and 1991; Antipenkoand Kuznetsov1998) and cinnamic glycosides in other countries it has been shown that human disturbance such as rosin, rosavin, and rosarin (Zapesochnaya and Kurkin for commercial purposes have reduced their genetic diversity 1982; Kurkin et al. 1986; Satsyperova et al. 1993) have been (Yan et al. 2003; Lei et al. 2006; Xia et al. 2007). One possi- well characterized. Other constituents of Rhodiola roots are ble solution could be the establishment of ex situ and in situ flavonoids, tannings and gallic acid and its esters. conservation of these plants to ensure the production of safe, 618 Pharmazie 65 (2010) ORIGINAL ARTICLES 100 A 90 80 /g 70 mg 60 ion at tr 50 en nc 40 Co 30 20 10 0 1471013161922252831343740434649525558616467707376798285889194 Roseroot clones 16 B 14 12 g g/ 10 nm io 8 at tr en 6 nc Co 4 2 0 14710 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70 73 76 79 82 85 88 91 94 Roseroot clones Fig. 1: Quantities of fivebioactive compounds produced by Norwegian roseroot determined by HPLC. All quantities are measured in mg g−1.A)The content of rosavin in 95 roseroot clones sorted based on increasing levels of rosavin. B) The content of tyrosol (red), salidroside (green), cinnamyl alcohol (yellow) and rosin (blue). The roseroot clones are listed in the same order as in A efficient and stable pharmaceutical products. In Norway a corresponding results from roseroot in other countries, and (iii) roseroot germplasm collection has been established and veg- investigate if genotypes producing high levels of bioactive com- etative cultivation of this plant has been successfully introduced pounds are genetically related and whether AFLP markers can be (Dragland 2001; Elameen et al. 2008). used to identify genetic resources of roseroot for further devel- The aims of this study were to: (i) identify and measure the pro- opment in future breeding programmes. Abreeding programme duction of the bioactive compounds rosavin, salidroside, rosin, in Norway followed by roseroot farming will be necessary in cinnamyl alcohol and tyrosol in 95 roseroot clones originating order to counteract the threat of human disturbance for com- from 15 different counties in Norway butgrown under uniform mercial purposes that otherwise might result in areduction or experimental conditions for three growing seasons; (ii) compare extinction of genetic diversity of natural roseroot populations. the chemical content of the Norwegian roseroot collection with Table 1: Quantities of the main bioactive compounds of 2. Investigation and results roseroot The chromatographic fingerprints from HPLC showed large qualitative and quantitative differences of the chemical con- Compound Minimum level Maximum level stituents in the R. rosea clones included in this study.The Rosavin 2.900 ± 0.006 85.950 ± 0.710 fivechemicals rosavin, salidroside, rosin, cinnamyl alcohol and Salidroside 0.030 ± 0.001 12.850 ± 0.001 tyrosol, were detected in all 95 roseroot clones, buthigh varia- Rosin 0.080 ± 0.001 4.750 ± 0.001 tion between the quantities of these compounds were revealed Cinnamyl alcohol 0.020 ± 0.001 1.180 ± 0.002 among the clones in the germplasm collection (Fig. 1A and 1B). Tyrosol 0.040 ± 0.001 2.150 ± 0.002 Single roseroot clones with high production of chemical com- pounds did not reflect acertain county of origin or plant gender. Values are presented in mg g−1 ± standard deviation Minimum and maximum levels of these bioactive compounds Pharmazie 65 (2010) 619 ORIGINAL ARTICLES Fig. 2: The relationships between the Norwegian roseroot clones determined by UPGMA cluster analysis based on their chemical production. Each roseroot clone waslabelled according to the code givenbyElameen et al. (2008) measured in the Norwegian roseroot clones were recorded 14.650, 5.480, 10.440 and 11.891 for rosavin, salidroside, rosin, (Table 1). cinnamyl alcohol and tyrosol. The relationships between the 95 clones revealed by UPGMA cluster analyses based on chemical production data are presented in Fig. 2. The UPGMA result did not reflect the county of origin of the clones. 3. Discussion Pearson correlation analysis showed significant and positive cor- In this study we describe the chemical characterization of a relations of the content of several of these bioactive compounds collection of Norwegian roseroot clones. As farasweknow produced in the roseroot clones analysed (Table 2). Best corre- this represents the largest collection of vegetatively propagated lation wasfound for the production of the important bioactive roseroot plants. compounds rosavin and salidroside (r =0.6194; P ≤ 0.01). A Except for cinnamyl alcohol, the maximum levels of the bioac- negative correlation wasfound between salidroside and rosin. tive compounds presented in this study were much higher than Forsome of the individual Norwegian clones the levelofchem- levels of bioactive compounds found in other Rhodiola species ical compounds exceeded the levelrequired for roseroot to be (Wang et al. 2006; Mao et al. 2007; Wiedenfeld et al. 2007). used for clinical treatment (Table 3). Differences in the content and composition of the essential Regression analysis showed alow,but not significant correla- chemical compounds of roseroot have been reported in several tion between the results from apreviously performed AFLP publications (e.g., Galambosi et al. 1999; Linh et al.