0003-6838/01/3703- $25.00 © 2001 MAIK “Nauka /Interperiodica” 0297 Applied Biochemistry and Microbiology, Vol. 37, No. 3, 2001, pp. 297–300. Translated from Prikladnaya Biokhimiya i Mikrobiologiya, Vol. 37, No. 3, 2001, pp. 344–348. Original Russian Text Copyright © 2001 by Sovetkina, Kalenik, Bulgakov, Yasnetskaya, Zhuravlev. Ginsenoids are the major biologically active sub- stances of ginseng [1]. The medicinal properties of these plants are also related to the presence of other physiological components, such as oligopeptides, amino acids, microelements, vitamins, and enzymes [2]. The importance of many microelements is beyond doubt. As, B, Cr, Co, Cu, F, I, Fe, Mn, Mo, Ni, Se, Si, V, and Zn are believed to be essential to living organ- isms [3]. The mineral composition of ginseng roots has received little attention [2]. Ginseng calluses were shown to contain Ag, Cu, Fe, Ge, Mg, Ni, and Zn [4]. We found no data on the mineral composition of callus cultures of other plants (SCI database). Lovkova et al. showed that alkaloid-producing medicinal plants accu- mulate considerable amounts of microelements that determine their properties [5]. This work was designed to evaluate the importance of ginseng cell cultures as sources of mineral elements, to compare the mineral compositions of natural ginseng roots and callus cultures, and to study a culture of trans- genic ginseng roots obtained in recent works. MATERIALS AND METHODS Materials. Roots of five-year-old Panax ginseng C. A. Mey plants were obtained from the Zhen’shen’ state farm (Varvarovka, Primorskii krai). Callus cultures of P. ginseng maintained in the Collection of Cell Cultures since their isolation in 1988 (Institute of Biology and Soil Science) [6] and transgenic roots 1c-rolC-II obtained from the line 1c in 1993 [7] were used. The following cultures were studied: shoots of two-month- old ginseng plantlets (1c, Mimaki; 2c, Japanese popu- lation; 4c, Korean population; and 7c and 7l, Primor- skaya populations), leaf plates of 2-year-old green- house plants, roots of two-month-old seedlings (7k, Primorskaya population), rhizome base of 17-year-old wild ginseng (10, Chuguevskii region, Primorskii krai), flower buds of eight-year-old wild ginseng (8ts, Ussuri- iskii region, Primorskii krai), and seedlings of Primor- skii ginseng obtained after transformation of Agrobac- terium rhizogenes A4 (R-1) [8]. Culturing conditions. Callus cultures and the culture of transgenic roots 1c-rolC-II were grown in 250-ml Erlenmeyer’s flasks with 60 ml of agar-treated W 4CPA and W IBA media, respectively, in the dark at 25°C and 50–70% relative humidity. The nutrient medium con- tained 400 mg/l NH 4 NO 3 , 1900 mg/l KNO 3 , 665 mg/l CaCl 2 · 6H 2 O, 170 mg/l MgSO 4 · 7H 2 O, 170 mg/l KH 2 PO 4 , 6.2 mg/l H 3 BO 3 , 22.3 mg/l MnSO 4 · 4H 2 O, 0.025 mg/l CoCl 2 · 6H 2 O, 0.025 mg/l CuSO 4 · 5H 2 O, 8.6 mg/l ZnSO 4 · 7H 2 O, 0.25 mg/l Na 2 MO 4 · 2H 2 O, 0.83 mg/l KJ, 22.3 mg/l FeSO 4 · 7H 2 O, 38 mg/l Na– EDTA, 100 mg/l mesoinositol, 100 mg/l peptone, 0.2 mg/l thiamine hydrochloride, 0.5 mg/l pyridoxine hydro- chloride, 0.5 mg/l nicotinic acid, 0.4 mg/l 4-chloropheny- lacetic acid, 25 000 mg/l sucrose, and 6000 mg/l agar [6]. The transformed roots were grown on a shaker at 80 rpm with an amplitude of 20–30 mm for 30 days. Elemental composition. The elemental composition was estimated, after incinerating with nitric acid vapors, by atom absorption spectroscopy on an ASH device (Nippon–Jarrel, Germany) [9]. The content of elements was expressed in μg/g dry weight. Data processing. The results were analyzed by using Statistica software (Windows 98). RESULTS AND DISCUSSION Importance of Ginseng callus cultures as a source of mineral elements. Table 1 shows the elemental com- positions of natural and transgenic ginseng roots and various callus cultures. The cultured ginseng cells dif- fered in their ability to accumulate mineral elements. The content of six of the eleven mineral components Mineral Composition of Cultured Ginseng Cells T. M. Sovetkina*, T. K. Kalenik*, V. P. Bulgakov**, E. G. Yasnetskaya**, and Yu. N. Zhuravlev** * Far-Eastern State Academy of Economics and Management, Vladivostok, 690019 Russia ** Institute of Biology and Soil Science, Far-Eastern Division, Russian Academy of Sciences, Vladivostok, 690022 Russia Received September 14, 2000 Abstract—The contents of macroelements and microelements in ginseng roots and callus cultures was deter- mined by atom absorption spectroscopy. Ginseng cells and tissues were shown to accumulate considerable amounts of microelements. The content of six of the eleven mineral components studied (K, Ca, Na, Mo, Mn, and Cr) in callus cultures was higher than that in roots of agricultural ginseng plants. We revealed good correlations between the contents of microelements (K, Ca, and Mg), as well as between the concentrations of macroele- ments (Mo, Li, Cu, and Cr), in ginseng cultures. The ability to accumulate elements varied between ginseng species, which was probably related to their genetic features. Our findings indicate that cultured ginseng cells hold much promise as a source of microelements.