The effect of arbuscular mycorrhiza on physiological and biochemical parameters and capsaicinoid production in Capsicum annuum L.: A comparative study of extraction methods and solvents




Mycorrhiza, Capsaicin, Ultrasound-assisted extraction, Funneliformis mosseae, Capsicum annuum, gas chromatography-mass spectrometry (GC-MS)


Paper description:

  • The aim of this study was to determine the effect of arbuscular mycorrhizal fungi on capsaicinoid production in Capsicum annuum, along with alterations in physiological and biochemical properties.
  • Magnetic stirring extraction and ultrasound-assisted extraction using three different solvents were used to determine capsaicinoids by GC-MS.
  • The highest capsaicin content up to 4-fold was found in plants inoculated with Funneliformis mosseae using ultrasound-assisted extraction with ethyl acetate. Plants with a mycorrhizal partner exhibited better physiological and biochemical properties.
  • Mycorrhizal fungi contribute to plant growth and development and enhance the production of valuable secondary metabolites in plants.

Abstract: This study aimed to determine the effect of two mycorrhizal fungi, Funneliformis mosseae (Fm) and Rhizophagus intraradices (Ri), on capsaicinoid production in Capsicum annuum L. by gas chromatography-mass spectrometry (GC-MS) via two different extraction approaches, magnetic stirring and ultrasound-assisted extraction with three different solvents, ethanol (EtOH), ethyl acetate (EtAce), and acetonitrile (AceN). The effect of mycorrhizal fungi on some physiological properties and biochemical activity, the content of total phenolic compounds, and antioxidant activity were also investigated. For all investigated parameters, the plants inoculated with mycorrhizal fungi showed significantly higher values than the non-mycorrhizal control plants, except for malondialdehyde (MDA), which was an indicator of lipid peroxidation due to damage that occurred in the cell membrane. It was concluded that inoculation with mycorrhizal fungi increased both capsaicin and dihydrocapsaicin production up to 4-fold in C. annuum compared to the control. Results also indicated that ultrasound-assisted extraction with EtAce was the most effective method for the determination of capsaicin by GC-MS.


Download data is not yet available.


Hamed M, Kalita D, Bartolo ME, Jayanty SS. Capsaicinoids, polyphenols and antioxidant activities of Capsicum annuum: Comparative study of the effect of ripening stage and cooking methods. Antioxidants. 2019;8(9):364-71.

Bosland PW. Chiles: history, cultivation and uses. In: Charalarnbous G, editor. Spices, Herbs and Edible Fungi. New York, USA: Elsevier Publication; 1994. p. 347-66.

Umamaheswai A, Lalitha V. In vitro effect of various growth hormones in Capsicum annum L. on the callus induction and production of Capsaicin. J Plant Sci. 2007;2(5):545-51. 10.3923/jps.2007.545.551

Pereira JAP, Vieira IJC, Freitas MSM, Prins CL, Martins MA, Rodrigues R. Effects of arbuscular mycorrhizal fungi on Capsicum spp. J Agric Sci. 2016;154(5):828-49.

Kumar OA, Tata SS. Ascorbic acid contents in chili peppers (Capsicum L.). Not Sci Biol. 2009;1(1):50-2.

Yilma G. The role of mycorrhizal fungi in pepper (Capsicum annuum) production. Int J Adv Res in Biol Sci. 2019;6(12):59-65.

Carlsen SCK, Understrup A, Fomsgaard IS, Mortensen AG, Ravnskov S. Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus. Plant and Soil. 2008;302:33-43.

Strack D, Fester T. Isoprenoid metabolism and plastid reorganization in arbuscular mycorrhizal roots. New Phytol. 2006;172(1):22-34.

Davies Jr FT, Potter JR, Linderman RG. Mycorrhiza and repeated drought exposure affect drought resistance and extraradical hyphae development of pepper plants independent of plant size and nutrient content. J Plant Physiol. 1992;139(3):289-94.

Hirrel MC, Gerdemann JW. Improved growth of onion and bell pepper in saline soils by two vesicular‐arbuscular mycorrhizal fungi. Soil Sci Soc Am J. 1980;44(3):654-55.

Bagyaraj DJ, Sreeramulu KR. Preinoculation with VA mycorrhiza improves growth and yield of chilli transplanted in the field and saves phosphatic fertilizer. Plant and Soil. 1982;69:375-81.

Berk Z. Food Process Engineering and Technology. Elsevier; 2009. 622 p.

Barbero GF, Liazid A, Azaroual L, Palma M, Barroso CG. Capsaicinoid contents in peppers and pepper-related spicy foods. Int J Food Prop. 2016;19(3):485-93.

Parrish M. Liquid chromatographic method for determining capsaicinoids in capsicums and their extractives: collaborative study. J Assoc Off Anal Chem. 1996;79(3):738-45.

Perucka I, Oleszek W. Extraction and determination of capsaicinoids in fruit of hot pepper Capsicum annuum L. by spectrophotometry and high-performance liquid chromatography. Food Chem.. 2000;71(2):287-91.

Peña-Alvarez A, Alvarado LA, Vera-Avila LE. Analysis of capsaicin and dihydrocapsaicin in hot peppers by ultrasound assisted extraction followed by gas chromatography-mass spectrometry. Instrum Sci Technol. 2012;40(5):429-40.

Phillips JM, Hayman DS. Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Br Mycol Soc. 1970;55(1):158-61.

Giovannetti M, Mosse B. An evaluation of techniques for measuring vesicular arbuscular mycorrhizal infection in roots. New Phytol. 1980:489-500.

Eisenback JD. A comparison of techniques useful for preparing nematodes for scanning electron microscopy. J Nematol. 1986;18(4):479-87.

McLean C, Lawrie AC. Patterns of root colonization in epacridaceous plants collected from different sites. Ann Bot. 1996;77(4):405-12.

Moran P, Coats B. Biological sample preparation for SEM imaging of porcine retina. Microscopy Today. 2012;20(2):28-31.

Lichtenthaler HK. Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Meth Enzymol. 1987;148:350-82.

Sumanta N, Haque CI, Nishika J, Suprakash R. Spectrophotometric analysis of chlorophylls and carotenoids from commonly grown fern species by using various extracting solvents. Res J Chem Sci. 2014;4(9):63-9.

Heath RL, Packer L. Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys. 1968;125(1):189-98.

Pascholati SF, Nicholson RL, Butler LG. Phenylalanine ammonia‐lyase activity and anthocyanin accumulation in wounded maize mesocotyls. J Phytopathol. 1986;115(2):165-72.

Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Meth Enzymol. 1999;299:152-78.

Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic. 1965;16(3):144-58.

Shimada K, Fujikawa K, Yahara K, Nakamura T. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem. 1992;40(6):945-48.

Peña-Alvarez A, Ramírez-Maya E, Alvarado-Suárez LÁ. Analysis of capsaicin and dihydrocapsaicin in peppers and pepper sauces by solid phase microextraction–gas chromatography–mass spectrometry. J Chromatogr A. 2009;1216(14):2843-47.

İslek Y. Assessment of oxidative stress in plants by EPR spectroscopy. In: Ozturk M, Bhat R, Ashraf M, Tonelli F, Unal B, Dar G, editors. Phytohormones and Stress Responsive Secondary Metabolites. Elsevier; 2023. p. 133-40.

Bagyaraj DJ. Microbial Biotechnology for Sustainable Agriculture, Horticulture & Forestry. New India Publishing Agency; 2011. 308 p.

Raghavendra Kumar M, Ashwin R, Bagyaraj DJ. Screening arbuscular mycorrhizal fungi in order to select the best for alleviating wilt disease complex of capsicum. Proc Natl Acad Sci India Sect B: Biol Sci. 2018;88:679-84.

Spoorthi VB, Ranadev P, Ashwin R, Bagyaraj JD. Response of Capsicum annuum L. Seedlings Raised in Pro Trays to Inoculation with AM Fungus Glomus bagyarajii and K Solubilizing Bacterium Frateuria aurantia. Seeds. 2022;1(4):315-23.

Boonlue S, Surapat W, Pukahuta C, Suwanarit P, Suwanarit A, Morinaga T. Diversity and efficiency of arbuscular mycorrhizal fungi in soils from organic chili (Capsicum frutescens) farms. Mycoscience. 2012;53(1):10-6.

Tanwar A, Aggarwal A, Panwar V. Arbuscular mycorrhizal fungi and Trichoderma viride mediated Fusarium wilt control in tomato. Biocontrol Sci Technol. 2013;23(5):485-98.

Russo VM, Perkins-Veazie P. Yield and nutrient content of bell pepper pods from plants developed from seedlings inoculated, or not, with microorganisms. HortScience. 2010;45(3):352-58.

Steinkellner S, Hage-Ahmed K, García-Garrido JM, Illana A, Ocampo JA, Vierheilig H. A comparison of wild-type, old and modern tomato cultivars in the interaction with the arbuscular mycorrhizal fungus Glomus mosseae and the tomato pathogen Fusarium oxysporum f. sp. lycopersici. Mycorrhiza. 2012;22:189-94.

Kapoulas N, Ilic ZS, Koukounaras A, Ipsilantis I. Application of arbuscular mycorrhizal inoculum in greenhouse soil with manure induced salinity for organic pepper production. Acta Sci Pol Hortorum Cultus. 2019;18(1):129–39.

Bulgarelli RG, Marcos FCC, Ribeiro RV, de Andrade SAL. Mycorrhizae enhance nitrogen fixation and photosynthesis in phosphorus-starved soybean (Glycine max L. Merrill). Environ and Exp Bot. 2017;140:26-33.

Wang J, Fu Z, Ren Q, Zhu L, Lin J, Zhang J, Cheng X, Ma J, Yue J. Effects of arbuscular mycorrhizal fungi on growth, photosynthesis, and nutrient uptake of Zelkova serrata (Thunb.) Makino seedlings under salt stress. Forests. 2019;10(2):186-202.

Çekiç FÖ, Ünyayar S, Ortaş İ. Effects of arbuscular mycorrhizal inoculation on biochemical parameters in Capsicum annuum grown under long term salt stress. Turk J Bot. 2012;36(1):63-72.

Abdel Latef AAH, Chaoxing H. Does inoculation with Glomus mosseae improve salt tolerance in pepper plants? J Plant Growth Regul. 2014;33:644-53.

Koç E, Üstün AS, İşlek C, Arıcı YK. Defence responses in leaves of resistant and susceptible pepper (Capsicum annuum L.) cultivars infected with different inoculum concentrations of Phytophthora capsici Leon. Sci Hortic. 2011;128(4):434-42.

Chitara MK, Keswani C, Varnava KG, Birla H, Dilnashin H, Singh SP, Sarojini V, Sperry J, Singh HB. Impact of the alkaloid colletotrichumine A on the pathogenicity of Colletotrichum capsici in Capsicum annum L. Rhizosphere. 2020;16:e100247.

Pereira LFP. Studies on phenylalanine ammonia-lyase and peroxidases in cassava, Manihot esculenta Crantz, during root post-harvest deterioration and interaction with Xanthomonas. Canada: University of Guelph; 1998. 191 p.

Sundaramoorthy S, Raguchander T, Ragupathi N, Samiyappan R. Combinatorial effect of endophytic and plant growth promoting rhizobacteria against wilt disease of Capsicum annum L. caused by Fusarium solani. Biol Control. 2012;60(1):59-67.

Ozgonen H, Yardimci N, Kilic HC. Induction of phenolic compounds and pathogenesis-related proteins by mycorrhizal fungal inoculations against Phytophthora capsici Leonian in pepper. Pak J Biol Sci. 2009;12(17):1181-87.

Fritz V, Tereucán G, Santander C, Contreras B, Cornejo P, Ferreira PAA, Ruiz A. Effect of inoculation with arbuscular mycorrhizal fungi and fungicide application on the secondary metabolism of Solanum tuberosum leaves. Plants. 2022;11(3):278-85.

Avio L, Sbrana C, Giovannetti M, Frassinetti S. Arbuscular mycorrhizal fungi affect total phenolics content and antioxidant activity in leaves of oak leaf lettuce varieties. Sci Hortic. 2017;224:265-71.

Bernardo VF, Garita SA, Arango MC, Ripodas JI, Saparrat MCN, Ruscitti MF. Arbuscular mycorrhizal fungi against the false root-knot nematode activity in Capsicum annuum: Physiological responses in plants. Biocontrol Sci Technol. 2021;31(2):119-31.

Banuelos J, Alarcón A, Larsen J, Cruz-Sánchez S, Trejo D. Interactions between arbuscular mycorrhizal fungi and Meloidogyne incognita in the ornamental plant Impatiens balsamina. J Soil Sci Plant Nutr. 2014;14(1):63-74.

Huang Z, Zou Z, He C, He Z, Zhang Z, Li J. Physiological and photosynthetic responses of melon (Cucumis melo L.) seedlings to three Glomus species under water deficit. Plant Soil. 2011;339:391-99.

da Luz SFM, Reis LdA, de Lemos OF, Maia JGS, De Mello AH, Ramos AR, Da Silva JKR. Effect of arbuscular mycorrhizal fungi on the essential oil composition and antioxidant activity of black pepper (Piper nigrum L.). Int J Appl Res Nat Prod. 2016;9(3):10-7.

Appendino G. Capsaicin and capsaicinoids. In: Fattorusso E, Taglialatela-Scafati O, editors. Modern Alkaloids: Structure, Isolation, Synthesis and Biology. Weinheim, Germany: Wiley; 2008. p. 73-109.

Kosuge S, Furuta M. Studies on the pungent principle of Capsicum: Part XIV Chemical Constitution of the Pungent Principle. Agric Biol Chem. 1970;34(2):248-56.

Juangsamoot J, Ruangviriyachai C, Techawongstien S, Chanthai S. Determination of capsaicin and dihydrocapsaicin in some hot chilli varieties by RP-HPLC-PDA after magnetic stirring extraction and clean up with C 18 cartridge. Int Food Res J. 2012;19(3):1217-26.

Chinn MS, Sharma-Shivappa RR, Cotter JL. Solvent extraction and quantification of capsaicinoids from Capsicum chinense. Food Bioprod Processing. 2011;89(4):340-45.

Abbas W, Ahmed D, Qamar MT. Surfactant-mediated extraction of capsaicin from Capsicum annuum L. fruit in various solvents. Heliyon. 2022;8(8):e10273.

Atique I, Ahmed D, Maqsood M, Malik W. Solvents for extraction of antidiabetic, iron chelating, and antioxidative properties from bottle gourd fruit. Int J Veg Sci. 2018;24(3):212-26.

Lu X, Zheng Z, Li H, Cao R, Zheng Y, Yu H, Xiao J, Miao S, Zheng B. Optimization of ultrasonic-microwave assisted extraction of oligosaccharides from lotus (Nelumbo nucifera Gaertn.) seeds. Ind Crops Prod. 2017;107:546-557.




How to Cite

Canpolat Şükrü, İşlek C. The effect of arbuscular mycorrhiza on physiological and biochemical parameters and capsaicinoid production in Capsicum annuum L.: A comparative study of extraction methods and solvents. Arch Biol Sci [Internet]. 2023Oct.26 [cited 2024Feb.29];75(3):327-39. Available from: