Callus formation and camphor aggregation in response to sorbitol stimulated osmotic stress in yarrow




Asteraceae, cell suspension cultures, elicitor, terpenoid


Sorbitol is an important source of abiotic stress that is used to increase osmolality in cell cultures. It increases the antioxidant enzymes of defense catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in the stress state of cells. Sorbitol plays an important role in stimulating these enzymes in cells and increasing phenylalanine ammonium lyase (PAL) activity. The aim of this study was to apply increasing doses of sorbitol elicitor to cell suspension cultures to determine the changes in cell number, viability, dry weight, and camphor content. In vitro plantlets were obtained from plant seeds and stem segments of these plants were used as explant source. Cell cultures were established after callus formation. Then, 0 (control), 5, 25, and 50 g L-1 sorbitol was dissolved in distilled water and cultured. Samples were taken three times in total, starting from day 1 to day 3. The content of camphor was detected by gas chromatography-mass spectrometry (GC-MS). Cell number, viability,dry weight, and camphor content increased significantly with increasing doses of sorbitol compared to sampling times. Compared to the initial culture, the amount of camphor increased by 40% at the 5 g L-1 dose, 82% at the 25 g L-1 dose, and 154% at the 50 g L-1 dose. In A. gypsicola cell cultures, increasing doses of sorbitol have clearly demonstrated the secondary metabolite accumulation and its positive effect on cell growth.

Author Biography

Ebru Batı Ay, Amasya University, Suluova Vocational School, Medicinal and Aromatic Plants Program, Amasya, Türkiye

Tıbbi ve aromatik bitkiler, antioksidanlar


Abdel-Rahman R, Alqasoumi S, El-Desoky A, Soliman G, Paré G, Hegazy ME. 2015. Evaluation of the anti-inflammatory, analgesic and anti-ulcerogenic potentials of Achillea fragrantissima (Forssk.). South African Journal of Botany, 98: 122-127.

Açıkgöz MA. 2020a. Establishment of cell suspension cultures of Ocimum basilicum L. and enhanced production of pharmaceutical active ingredients. Ind Crops Prod 148:112278.

Açıkgöz MA. 2020b. Determination of essential oil compositions and antimicrobial activity of Achillea gypsicola Hub.-Mor. at different plant parts and phenological stages. Journal of Essential Oil Research, 32(4), 331-347.

Açıkgöz MA. 2021. Effects of sorbitol on the production of phenolic compounds and terpenoids in the cell suspension cultures of Ocimum basilicum L. Biologia, 76(1), 395-409.

Açıkgöz MA. 2017. Achillea gypsicola türünde kallus kültürü ile sekonder metabolit üretim potansiyelinin belirlenmesi. Ordu University (Doctoral dissertation, PhD Thesis (Printed), Ordu).

Açıkgöz MA, Kara ŞM. 2019. Effect of various pretreatments on germination of Turkish endemic Achillea gypsicola Hub.-Mor. species under in vivo and in vitro conditions. Journal of the Institute of Science and Technology, 9(4), 2321-2329.

Açıkgöz MA, Ebru BA, Aygün A, Kara ŞM. 2022. Light-Mediated Biosynthesis of Phenylpropanoid Metabolites in Cell Suspension Cultures of Turkish Yarrow (Achillea gypsicola Hub. Mor.). Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 25(6), 1234-1242

Açıkgöz MA, Ebru BA, Kara ŞM, Aygün A. 2023. Influence of Selected Heavy Metals on Cell Growth and Camphor Secretion in Achillea gypsicola Hub. Mor. In vitro Cell Cultures. Journal of Agricultural Faculty of Gaziosmanpaşa University (JAFAG), 40(1), 11-18.

Açıkgöz MA, Kara ŞM, Ebru BA, Odabaş S. 2018b. Effect of light on biosynthesis of alkamide, caffeic acid derivatives and echinacoside in Echinacea purpurea L. callus cultures. Akademik Ziraat Dergisi, 7(2), 179-184.

Açıkgöz MA, Yarılgaç T, Kara ŞM. 2018a. Enhancement of phytochemical compounds using biotic and abiotic elicitors in purple coneflower (Echinacea purpurea L.). Indian J Pharm Educ Res, 52, 140-145.

Açıkgöz MA, Kara SM, Aygun A, Ozcan MM, Ay EB. 2019. Effects of methyl jasmonate and salicylic acid on the production of camphor and phenolic compounds in cell suspension culture of endemic Turkish yarrow (Achillea gypsicola) species. Turkish Journal of Agriculture and Forestry

Açıkgöz MA. 2019. Evaluation of phytochemical compositions and biological properties of Achillea gypsicola at different phenological stages. Chemistry & Biodiversity, 16(12), e1900373.

Ahmadi-Dastgerdi A, Ezzatpanah H, Asgary S, Dokhani S, Rahimi E. 2017. Phytochemical, Antioxidant and Antimicrobial Activity of the Essential Oil from Flowers and Leaves of Achillea millefolium subsp. millefolium. Journal of Essential Oil Bearing Plants, 20(2): 395-409.

Al-Khayri JM, Al-Bahrany AM. 2002. Callus growth and proline accumulation in response to sorbitol and sucrose-induced osmotic stress in rice. Biol Plant 45:609–611.

Almadiy AA, Nenaah GE, Al Assiuty BA, Moussa EA, Mira NM. 2016. Chemical composition and antibacterial activity of essential oils and major fractions of four Achillea species and their nanoemulsions against foodborne bacteria. LWT-Food Science and Technology, 69: 529-537.

Azarabadi S, Abdollahi H, Torabi M, Salehi Z, Nasiri J. 2017. ROS generation, oxidative burst and dynamic expression profiles of ROS-scavenging enzymes of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in response to Erwinia amylovora in pear (Pyrus communis L). Eur J Plant Pathol 147:279–294.

Bidabadi SS, Meon S, Wahab Z, Subramaniam S, Mahmood M. 2012. In vitro selection and characterization of water stress tolerant lines among ethyl methanesulphonate (EMS) induced variants of banana (Musa spp., with AAA genome). Australian Journal of Crop Science, 6(3), 567-575.

Cheng LH, Eugene ICW, Yu CS. 2009. Essential Oil Compositions and Bioactivities of the Various Parts of Cinnamomum camphora Sieb.var, linaloolifera Fujuta, Journal of Forestry Research, 2: 77-96.

Chong TM, Abdullah MA, Lai OM, Nor’Aini FM, Lajis NH. 2005. Effective elicitation factors in Morinda elliptica cell suspension culture.

Dağlioğlu Y, Açıkgöz MA, Özcan MM, Kara ŞM. 2022. Impact of application of alumina oxide nanoparticles on callus induction, pigment content, cell damage and antioxidant enzyme activities in Ocimum basilicum. Journal of International Environmental Application and Science, 17(1), 22-33.

de Costa F, Yendo ACA, Fleck JD, Gosmann G, Fett-Neto AG. 2013. Accumulation of a bioactive triterpene saponin fraction of Quillaja brasiliensis leaves is associated with abiotic and biotic stresses. Plant Physiol Biochem 66:56–62.

Demirci B, Başer KHC, Aytaç Z, Khan SI, Jacob MR, Tabanca N. 2017. Comparative Study of Three Achillea Essential Oils from Eastern Part of Turkey and their Biological Activities.

Dodds JH, Roberts LW. 1985. Experiments in plant tissue culture. International Potato Center.

Ebru BA, Kocaman B, Öner EK, Açıkgöz MA. 2022. Effects of elicitors on secondary metabolite (SM) production and antioxidant activity in sweet basil (Ocimum basilicum L.) cell suspension cultures. Notulae Scientia Biologicae, 14(2), 11246-11246.

Frizzo CD, Santos AC, Paraul N, Serafini LA, Dellacassa E, Lorenzo D, Moyna P. 2000. Essential Oils of Camphor Tree (Cinnamomum camphora Nees and Eberm) Cultivated in Southern Brazil. Journal of Brazilian Archives of Biology and Technology, 43(3): 313-316.

Ghasemi Pirbalouti A. 2017. Chemical Composition of the Essential Oils from the Leaves and Flowers of Two Achillea species from Iran. Journal of Essential Oil Bearing Plants, 20(1): 205-214.

Hong MLK, Bhatt ARVIND, Ping NS, Keng CL. 2012. Detection of elicitation effect on Hyoscyamus niger L. root cultures for the root growth and production of tropane alkaloids. Rom Biotech Lett 17:7340–7351

Kang SM, Min JY, Kim YD, Karigar CS, Kim SW, Goo GH, Choi MS. 2009 Effect of biotic elicitors on the accumulation of bilobalide and ginkgolides in Ginkgo biloba cell cultures. J Biotechnol 139:84–88.

Khiyari MEA, Kasrati A, Jamali CA, Zeroual S, Markouk M, Bekkouche K, Abbad A. 2014. Chemical composition, antioxidant and insecticidal properties of essential oils from wild and cultivated Salvia aucheri subsp. blancoana (Webb. & Helder)), an endemic, threatened medicinal plant in Morocco. Industrial Crops and Products, 57: 106-109.

Kubeš J, Tůmová L, Martin J, Vildová A, Hendrychová H, Sojková K. 2014 The production of isoflavonoids in Genista tinctoria L. cell suspension culture after abiotic stressors treatment. Nat Prod Res 28:2253–2263.

Laloue M, Courtois D, Manigault P. 1980 Convenient and rapid fluorescent staining of plant cell nuclei with ‘33258’Hoechst. Plant Sci Lett 17:175–179.

Lin TC, Chu FH, Tseng YH, Tsai JB, Chang ST, Wang SY. 2007. Bioactivity Investigation of Lauraceae Trees Grown in Taiwan. Pharmaceutical Biology, 8: 638-644.

Ling OS, Kiong ALP, Hussein S. 2008. Establishment and optimisation of growth parameters for cell suspension cultures of Ficus deltoidea. American-Eurasian Journal of Sustainable Agriculture, 2(1): 38-49.

Moroff G, Eich J, Dabay M. 1994. Validation of use of the nageotte: hemocytometer to count low levels of white cells in white cell reduced platelet components. Transfusion, 34: 35-38.

Namdeo AG. 2007. Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev 1:69–79

Nazir M, Tungmunnithum D, Bose S, Drouet S, Garros L, Giglioli-Guivarc’h N, Abbasi BH, Hano C. 2019. Differential production of phenylpropanoid metabolites in callus cultures of Ocimum basilicum L. with distinct in vitro antioxidant activities and in vivo protective effects against UV stress. J Agric Food Chem 67:1847–1859.

Patil JG, Ahire ML, Nitnaware KM, Panda S, Bhatt VP, Kishor Kavi PB, Nikam TD. 2013. In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding. Applied microbiology and biotechnology, 97(6): 2379-2393.

Pavlović M, Petrović S, Milenković M, Kukić J, Couladis M, Tzakou O, Niketić M. 2008. Composition and antiradical capacity of Achillea grandifolia essential oil from Serbia. Planta Medica, 74(9): 1191-1191.

Placide R, Carpentier S, Swennen R. 2012. Development of in vitro technique to screen for drought tolerant banana varieties by sorbitol induced osmotic stress. African Journal of Plant Science, 6(15), 416-425. Process Biochem 40:3397–3405

Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan AK. 2011. Developing stress tolerant plants through in vitro selection- an overview of the recent progress. Environmental and Experimental Botany, 71(1), 89-98.

Ramulifho E, Goche T, Van As J, Tsilo TJ, Chivasa S, Ngara R. 2019. Establishment and characterization of callus and cell suspension cultures of selected Sorghum bicolor (L.) Moench varieties: a resource for gene discovery in plant stress biology. Agronomy 9:218.

Razavizadeh R, Adabavazeh F, Chermahini MR. 2017. Adaptive Responses of Carum copticum to in vitro Salt Stress. World Academy of Science, Engineering and Technology, International Journal of Biological, Biomolecular, Agricultural, Food and Biotechnological Engineering, 11(1): 37-42.

Salehi M, Karimzadeh G, Naghavi MR. 2019 Synergistic effect of coronatine and sorbitol on artemisinin production in cell suspension culture of Artemisia annua L. cv. Anamed. Plant Cell Tissue Organ Cult 137:587–597.

Sampietro DA, Lizarraga EF, Ibatayev ZA, Omarova AB, Suleimen YM, Catalán CA. 2016. Chemical composition and antimicrobial activity of essential oils from Acantholippia deserticola, Artemisia proceriformis, Achillea micrantha and Libanotis buchtormensis against phytopathogenic bacteria and fungi. Natural product research, 30(17): 1950-1955.

Sarmadi M, Karimi N, Palazón J, Ghassempour A, Mirjalili MH. 2019. Improved effects of polyethylene glycol on the growth, antioxidative enzymes activity and taxanes production in a Taxus baccata L. callus culture. Plant Cell Tissue Organ Cult 137:319–328.

Sherkheli MA, Benecke H, Doerner JF. 2009. Monoterpenoids induce agonist-specific desensitization of transient receptor potential vanilloid-3 (TRPV3) ion channels, Journal Pharmaclogy Science, 12: 116-128.

Singh B, Sahu PM, Sharma RA. 2017. Effect of elicitors on the production of pyrroloquinazoline alkaloids by stimulating anthranilate synthase activity in Adhatoda vasica Nees cell cultures. Planta, 1-13.

Smelcerovic A, Lamshoeft M, Radulovic N, Ilic D, Palic R. 2010. LC–MS Analysis of the Essential Oils of Achillea millefolium and Achillea crithmifolia. Chromatographia, 71(1-2): 113-116.

Sytar O, Kumar A, Latowski D, Kuczynska P, Strzałka K, Prasad MNV. 2013. Heavy metal-induced oxidative damage, defense reactions, and detoxification mechanisms in plants. Acta Physiol Plant 35:985–999.

Thompson MR, Douglas TJ, Obata‐Sasamoto H, Thorpe TA. 1986. Mannitol metabolism in cultured plant cells. Physiologia Plantarum, 67(3), 365-369.

Toncer O, Basbağ S, Karaman S, Diraz E, Basbağ M. 2010. Chemical composition of the essential oils of some Achillea species growing wild in Turkey. International Journal of Agriculture and Biology, 12(4): 527-530.

Valayıl JM, Kuriakose GC, Jayabaskaran C. 2015. Modulating the biosynthesis of a bioactive steroidal saponin, cholestanol glucoside by Lasiodiplodia theobromae using abiotic stress factors. International Journal of Pharmacy and Pharmaceutical Sciences, 7: 114-117.

Vanhove AC, Vermaelen W, Panis B, Swennen R, Carpentier, SC. 2012. Screening the banana biodiversity for drought tolerance: can an in vitro growth model and proteomics be used as a tool to discover tolerant varieties and understand homeostasis. Frontiers in Plant Science, 3, 176.

Vuleta A, Jovanović SM, Tucić B. 2016. Adaptive flexibility of enzymatic antioxidants SOD, APX and CAT to high light stress: the clonal perennial monocot Iris pumila as a study case. Plant Physiol Biochem 100:166–173.

Wu JY, Shi M. 2008. Ultrahigh diterpenoid tanshinone production through repeated osmotic stress and elicitor stimulation in fed-batch culture of Salvia miltiorrhiza hairy roots. Applied Microbiology and Biotechnology, 78(3): 441-448.

Yang Y, Pu X, Qu X, Chen F, Zhang G, Luo Y. 2017. Enhanced production of camptothecin and biological preparation of N1-acetylkynuramine in Camptotheca acuminata cell suspension cultures. Applied microbiology and biotechnology, 101(10): 4053-4062.

Zaker A, Sykora C, Gössnitzer F, Abrishamchi P, Asili J, Mousavi SH, Wawrosch C. 2015. Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel. Industrial Crops and Products, 67: 97-102.

Zhao JL, Zhou LG, Wu JY. 2010. Effects of biotic and abiotic elicitors on cell growth and tanshinone accumulation in Salvia miltiorrhiza cell cultures. Applied Microbiology and Biotechnology, 87(1): 137-144.




How to Cite

Açıkgöz, M. A., Aygün, A., Batı Ay, E., & Kara, Şevket M. (2023). Callus formation and camphor aggregation in response to sorbitol stimulated osmotic stress in yarrow. Turkish Journal of Agriculture - Food Science and Technology, 11(10), 1882–1888.



Research Paper