Effect of Different Levels of Sulfur on Growth, Quality Characteristics and Yield of Arnica chamissonis Less. ssp. foliosa

Document Type : Research Article


1 Shahid Beheshti University

2 Trabiat Modares University


Introduction: The genus Arnica L. comprises of 32 species predominantly confined to the boreal and montane region of the northern hemisphere. Arnica species are rhizomatous perennial herbs belonging to the daisy family, with simple or branched stems bearing opposite leaves, and large, single or cymose heads of yellow flowers. A. chamissonis is distributed over North America from Alaska to New Mexico, and due to its low ecological demand it is easier to cultivate than A. montana. Arnica is a source of sesquiterpene lactones, flavonoids, essential oils, terpenoids, and phenolic acids and exhibits antiseptic, anti-inflammatory, antiradical, antibacterial, anti-sclerotic, antifungal, and antioxidant activities. The flower heads and other parts of the plant of two of the species have been used therapeutically: A. montana and A. chamissonis, both of them containing sesquiterpene lactones as pharmacologically active compounds. Arnica species are used in as many as 300 drug preparations in Europe and about 20 products in Canada. Moreover, A. chamissonisis a good source of bioactive compounds a valuable source of herbal raw material and a pharmaceutical substitute for the endangered mountain arnica.  A. chamissonis extracts exhibit potent anti-inflammatory and anti-radical activity and possesses high antioxidant abilities that might be helpful in preventing or slowing the progress of free radical-dependent diseases. Low pH soil is one of the principal ecological requirements of Arnica. Good plant growth and flower yield was achieved on acid soils with a pH of 6.8 and below. After nitrogen, phosphorus and potassium, sulfur is considered as fourth major element in most crops. Rate of sulfur oxidation in soils vary and depend on population Thiobacillus bacteria in soil, particle size and environmental conditions.
Materials and Methods: This study was conducted as a complete randomized block design with three replications and four treatments including different levels of bentonite sulfur (0, 250, 500 and 750 kg/ha) combined with the bacterium Thibacillus thiooxidans. Traits such as height and width of plants, number of flowers on each plant, inflorescence diameter, fresh and dry yield of flowers in each plant per square meter and total phenols, total flavonoids, rutin, luteolin and apigenin content in flowers were measured. High-Performance Liquid Chromatography method (HPLC)  was used to separate rutin, luteolin and apigenin. HPLC grade methanol and distilled water, each with 0.02% added TFT were used as solvents.
Results and Discussion: The results showed that A. chamissonis is compatible with Tehran’s climate and it can be cultivated in regions with a similar climate. The use of elemental sulfur has a significant role in reducing soil pH and soil pH decreased more rapidly with increasing sulfur. Thiobacillus bacteria by oxidation of sulfur produced some to sulfuric acid and at low buffered properties can considerably reduce pH. Different levels of sulfur fertilizer had a significant effect (P ≤0.01) on soil acidity (pH), electrical conductivity, and sulfur, iron, zinc and manganese levels. The use of 750 kg sulfur bentonite with Thiobacillus caused to reduce soil acidity by about 0.9 units, and it increased the electrical conductivity of the soil to 7.33 dS/m. The amount of soil sulfate as a result of oxidation of sulfur fertilizers increased linearly. The highest amounts of iron, zinc and manganese were measured in the treatment of 750 kg sulfur as 3.38, 3.84 and 27.94 mg /kg, respectively. In this study, supplying 250 kg of sulfur plus bio-sulfur caused to improve the morphological traits and the highest flower yield (20.67 g / plant) compared to control (3.66 g / plant). Also, it increased the amount of sulfur and micro-elements in the soil, but in the treatment of 500 and 750 kg sulfur per hectare due to increased electrical conductivity and soil salinity, the growth of Chamisso Arnica decreased. Therefore, these levels of sulfur are not suitable for this plant. Sulfur Bentonite is oxidized by Thiobacillus bacteria and other microorganisms and sulfuric acid is produced. Reducing acidity and, as a result, the release of stabilized and insoluble nutrients can increase their uptake capacity and improve plant growth and development. Due to the increased salinity and osmotic imbalance in the soil by the treatments of 500 and 750 g /kg sulfur, vegetation growth will be incomplete and plants quickly enter to the reproductive phase and produce fewer and smaller flowers. The highest content of rutin was measured in plants grown in soil treated with 750 Kg/ha sulfur while soil treatment with 500 Kg/ha sulfur resulted in highest content of luteolin and apigenin, both showing significant difference to the control.
Conclusion: Results of this study indicated that the Chamisso Arnica showed adaptability to the planting location and it can be mass-cultivated under similar conditions.


1- Akula, R. and Ravishankar, G. A. 2011. Influence of abiotic stress signals on secondary metabolites in plants. Plant Signaling and Behavior, 6: 1720-1731.
2- Bahreininejad, B., Razmjou, J., and Mirza, M. 2013. Influence of water stress on morpho-physiological and phytochemical traits in Thymus daenensis. International Journal of Plant Production, 7(1): 151-166.
3- Bailly, C. 2004. Active oxygen species and antioxidants in seed biology. Seed Science Research, 14: 93-107.
4- Caliskan, O., Radusiene, J., Temizel, K.E., Staunis, Z., Cirak, C., Kurt, D., and Odabas, M.S. 2017. The effects of salt and drought stress on phenolic accumulation in greenhouse-grown Hypericum pruinatum. Italian Journal of Agronomy, 12(3): 271-275.
5- Dash, S. and Gupta, N. 2011. Microbial bioinoculants and their role in plant growth and development. International Journal of Biotechnology and Molecular Biology Research, 2: 232-251.
6- Dawood, F. A., Al-Omari, S. M., & Murtadha, N. S. 1985. High levels of sulfur affecting availability of some micronutrients in calcareous soils (in Iraq). J. of Agriculture and Water Resources Research (Iraq).‏
7- Delabays, N. and Mange, N. 1991. La culture d' Arnica montana L: aspects agronomiques et phytosanitaires. Revue suisse de viticulture, arboriculture, horticulture.
8- Doroudian, H.R., BesharatiKelayeh, H., FallahNosrat Abad, A.R., Heidary Sharif Abadi, H., Darvish, F., and Allahverdi, A. 2010. Study of absorbable phosphorus changes in lime soils and its impact on corn yield. Agricultural Modern Knowledge (Modern Knowledge of Sustainable Agriculture) 6(18): 27-35. (In Persian with English Summary).
9- El-Din, A.E., Aziz, E.E., Hendawy, S.F., and Omer, E.A. 2009. Response of Thymus vulgaris L. to salt stress and alar (B9) in newly reclaimed soil. Journal of Applied Sciences Research, 5: 2165-2170.
10- Esra, K.O.Ç., İŞLEK, C., and Üstün, A.S. 2010. Effect cold on protein, proline, phenolic compounds and chlorophyll content of two pepper (Capsicum annuum L.) varieties. Gazi University Journal of Science, 23(1): 1-6.
11- Finch C., Grant, G., Patersons, J., and Extension, B. C. 2004. Sulfur and soil pH, www.plantansewers.com, garden column.
12- Garcia-gomez, A., Bernal, M., and Roig, A. 2002. Growth of ornamental plants in two composts prepared from agroindustrial wastes. Bioresource Technology, 83: 81-87.
13- Gaspar, A., Cracinescu, O., Trif, M., Moisei, M., and Moldovan, L. 2014. Antioxidantand anti-inflammatory properties of active compounds from Arnica montana L. Romanian Biotechnological Letters 19 (3), 9353-9365.
14- Gawlik-Dziki, U.,´Swieca, M., Sugier, D., and Cichocka, J. 2009. Seeds of Arnica montana and Arnica chamissonis as a potential source of natural antioxidants. HerbaPolonica 55, 60-71.
15- Goodarzi, K. and Hosseini Farahi, M. 2014. Evaluating the ability of sulfur and animal manure to relieve Fe, Mn, Zn, Cu and B deficiency in 'Seah' table grapes in Cisakht region of Iran. I International Symposium on Organic Matter Management and Compost Use in Horticulture, 1018: 287-291.
16- Grant, C., Clayton, G., and Johnston, A. 2003. Sulphur fertilizer and tillage effects on canola seed quality in the black soil zone of western Canada. Canadian Journal of Plant Science, 83: 745-758.
17- Hitsuda, K., Yamada, M., and Klepker, D. 2005. Sulfur requirement of eight crops at early stages of growth. Agronomy Journal, 97: 155-159.
18- Hulten, E. 1968. Flora of Alaska and neighboring territories: a manual of the vascular plants. Stanford University Press, USA, pp. 920-921.
19- Janzen, H. and Bettany, J. 1987. The effect of temperature and water potential on sulfur oxidation in soils. Soil Science, 144: 81-89.
20- Kalbas, M., Filsoof, F., and Rezai‐nejad, Y. 1988. Effect of sulfur treatments on yield and uptake of Fe, Zn, and Mn by corn, sorghum, and soybeans. Journal of Plant Nutrition, 11: 1353-1360.
21- Kamtekar, S., Keer, V., and Patil, V. 2014. Estimation of phenolic content, flavonoid content, antioxidant and alpha amylase inhibitory activity of marketed polyherbal formulation. Journal of Applied Pharmaceutical Science, 4(9): 61-65.
22- Khavazi, K., Nourgholipour, F., and Malakouti, M.J. 2001. Effect of thiobacillus and phosphate solubilizing bacteria on increasing P availability from rock phosphate for corn. In International Meeting on Direct Application of Rock Phosphate and Related Appropriate Technology Latest Development and Practical Experience. Kuala Lumpur, Malaysia. 4(5):330-334
23- Kowalski, R., Sugier, D., Sugier, P., and Kołodziej, B. 2015. Evaluation of the chemical composition of essential oils with respect to the maturity of flower heads of Arnica montana L. and Arnica chamissonis Less. cultivated for industry. Industrial Crops and Products, 76: 857-865.
24- Lawrence, J. and Germida, J. 1988. Relationship between microbial biomass and elemental sulfur oxidation in agricultural soils. Soil Science Society of American Journal, 52: 672-677.
25- Li, X. S., Sato, T., Ooiwa, Y., Kusumi, A., Gu, J. D., and Katayama, Y. 2010. Oxidation of elemental sulfur by Fusarium solani strain THIF01 harboring endobacterium Bradyrhizobium sp. Microbial Ecology, 60: 96-104.
26- Loeppert, R. H. and Suarez, D. L. 1996. Carbonate and gypsum. Publications from USDA-ARS / UNL Faculty, 5: 437-474.
27- Maguire, B. 1943. A monograph of the genus Arnica. Brittonia, 4: 386-510.
28- Malakouti, M.J. 2004. Soil Fertility of Arid and Semi-Arid Regions, Tarbiat Modares University Press, 482 pp.
29- Malakouti, M.J., Baybordi, A., Lotfolahi, M., Shahabi, A., Siavoshi, K., Vakil, R., Ghaderi, J., Shahabifar, J., Majidi, A., and Jafarnejadi, A. 2010. Comparison of complete and sulfur coated urea fertilizers with pre-plant urea in increasing grain yield and nitrogen use efficiency in wheat. Journal of Agricultural Science and Technology, 10: 173-183.
30- Merfort, I. 2003. Arnica: new insights on the molecular mode of action of a traditional medicinal plant. Forschende Komplementarmedizin und klassische Naturheilkunde= Research in complementary and natural classical medicine, 10: 45-48.
31- Modaihsh, A.S., Al-Mustafa W. A., and E., Metwallv A. 1989. Effect of elemental sulfur on chemical changes and nutrient availability in calcareous soils. Plant and Soil, 116: 95-101.
32- Mohammadi Torkashvand, A. 2011. Effect of steel converter slag as iron fertilizer in some calcareous soils. Acta Agriculturae Scandinavica Section B-Soil and Plant Science, 61: 14-22.
33- Radanovic, D., Pljevljakusic, D., Markovic, T., Ristic, M., Dragoja, R., Pljevljakusic, D. and Markovic, T. 2007. Influence of fertilization model and PE mulch on yield and quality of Arnica (A. montana) at dystric cambisol. Zemljište i biljka, 56: 85-95.
34- Salehi, P., Asghari, B., Esmaeili, M. A., Dehghan, H., and Ghazi, I. 2013. α-Glucosidase and α-amylase inhibitory effect and antioxidant activity of ten plant extracts traditionally used in Iran for diabetes. Journal of Medicinal Plants Research, 7: 257-266.
35- Sangtarash, M.H., Qaderi, M.M., Chinnappa, C.C., and Reid, D.M. 2009. Differential sensitivity of canola (Brassica napus) seedlings to ultraviolet-B radiation, water stress and abscisic acid. Environmental and Experimental Botany, 66(2): 212-219.
36- Seyoum, A., Asres, K., and El-Fiky, F.K. 2006. Structure–radical scavenging activity relationships of flavonoids. Phytochemistry, 67(18), : 2058-2070.
37- Singh, R., Bhumbla, D., and Keefer, R. 1995. Recommended soil sulfate-S tests. Recommended soil testing procedures for the Northeastern United States. Northeast Regional Bulletin, 493: 46-51.
38- Small, E. and Catling, P. M. 2000. Les cultures medicinales canadiennes, NRC Research Press.
39- Smallfield, B. and Douglas, M. 2008. Arnica montana a grower’s guide for commercial production in New Zealand. A Report Prepared for New Zealand Arnica Growers’ Group, New Zealand Institute for Crop and Food Research Limited, Christchurch, New Zealand, December, 2.
40- Sorokin, D. Y. 2003. Oxidation of inorganic sulfur compounds by obligately organotrophic bacteria. Microbiology, 72: 641-653.
41- Sparks, D. L., Page, A., Helmke, P., Loeppert, R., Soltanpour, P., Tabatabai, M., Johnston, C., and Sumner, M. 1996. Methods of soil analysis. Part 3-Chemical methods, Soil Science Society of America Inc. 1309 pp.
42- Stursa, J., Kwiatkowski, P., Harcarik, J., Zahradnikova, J., and Krahulec, F. 2009. Cerný a cervený seznam cevnatých rostlin Krkonos/Black and red list of vascular plants of the Krkonose/Karkonosze Mts. (Western Sudetes CZ, PL). Opera Corcontica, 46: 67-104.
43- Sugier, D. 2007. The flowering pattern of Arnica montana L. and A. chamissonis Less. Under field cultivation conditions with successive flower head collection. Acta Agrobotanica, 60 (2): 133-139.
44- Sugier, D. 2013. Yield and chemical composition of mountains arnica (Arnica montana L.) raw material in relation to the method of plantation establishment andthe harvesting time of flower heads. Annales UMCS Agricultura 68 (3), 51-62.
45- Szulc, P., Waligora, H., and Skrzypczak, W. 2008. Better effectiveness of maize fertilization with nitrogen through additional application of magnesium and sulphur. Nauka Przyroda Technologie. Uniwersytet Przyrodniczy w Poznaniu, 2 (3):art. 19.
46- Terry, N. 1976. Effects of sulfur on the photosynthesis of intact leaves and isolated chloroplasts of sugar beets. Plant Physiology, 57: 477-479.
47- Valifard, M., Mohsenzadeh, S., Kholdebarin, B., and Rowshan, V. 2014. Effects of salt stress on volatile compounds, total phenolic content and antioxidant activities of Salvia mirzayanii. South African Journal of Botany, 93:.92-97.
48- Weremczuk-Jeżyna, I., Kisiel, W., and Wysokińska, H. 2006. Thymol derivatives from hairy roots of Arnica montana. Plant Cell Reports, 25: 993-996.
49- Willuhn, G. 1972. Studies on the contents of Arnica species. V. content and variations in content of volatile oils in the various organs of Arnica species. Planta Medica, 21: 221-245.
Volume 33, Issue 2 - Serial Number 2
November 2019
Pages 219-232
  • Receive Date: 03 April 2018
  • Revise Date: 03 December 2018
  • Accept Date: 08 April 2019
  • First Publish Date: 23 August 2019