Document Type : Research paper


1 Department of Plant Ecophysiology, Faculty of Agriculture, University of Tabriz, Iran

2 Department of Plant Physiology, Faculty of Natural science, University of Tabriz, Iran

3 Department of Soil Science, Faculty of Agriculture, University of Tabriz, Iran


The present study was carried out to investigate biochemical responses of marigold (Calendula officinalis L.) to Bio-organic fertilizers. Effects of co-inoculation of two plant growth-promoting rhizobacteria (PGPR) (Azotobacter sp.145PI and Azospirillum sp.AC49I), humic acid (HA) (10 kg ha−1), vermicompost (VC) (7 T ha−1) and combinations of these treatments were evaluated in two marigold cultivars [Isfahan double flower (DF) and single flower (SF)]. The biosynthesis of leaf protein, soluble and insoluble carbohydrate contents were increased in treated plants, while total free amino acid content was not influenced by treatments. Fertilizers also had positive impact on leaf and flower pigments, total flavonoid content, and total phenolic contents. Maximum amounts of anthocyanins were obtained in the DF cultivar treated by VC+PGPR and VC, which were 11.414 and 11.192 µmol g-1 FW, respectively. The SF cultivar treated by PGPR (36.11 %) and also the same cultivar treated by VC (33.39 %) had the highest antioxidant activities. In general, simultaneous application of fertilizers and also vermicompost were recognized as the best treatment for marigold plants. In conclusion, the findings of the current study confirmed that chemical composition and antioxidant activity of marigold can be positively influenced by Bio-organic fertilizers; therefore they can be used to obtain medicinal plants with improved quality during sustainable agricultural practices.


Abdelaziz M, Pokluda R, Abdelwahab M. 2007. Influence of compost, microorganisms and NPK fertilization upon growth, chemical composition and essential oil production of rosmarinus officinalis L. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 35, 86-90.

Aliniaeifard S, Seif M, Arab M, Zare Mehrjerdi M, Li T, Lastochkina O. 2018. Growth and Photosynthetic Performance of Calendula officinalis under Monochromatic Red Light. International Journal of Horticultural Science and Technology 5(1), 123-132.

Ansari R.A, Rizvi R, Sumbul A, Mahmood I. 2017. PGPR: Current Vogue in Sustainable Crop Production. In: Probiotics and Plant Health. eds V. Kumar, M. Kumar, S. Sharma, Prasad R. Singapore, Springer Publisher, 455-472.

Baskaran K. 2017. Pharmacological Activities of Calendula officinalis. International Journal of Science and Research 6(5), 43-47.

Bradford M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Analytical Biochemistry 72, 248-259.

Chang C, Yang M, Wen H, Chern J. 2002. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of FoodandDrug Analysis 10, 178-182.

Chen Y, Aviad T. 1990. Effects of humic substances on plant growth. In: MacCarthy P, Clapp C.E, Malcom R.L, Bloom P.R. Humic Substances in Soils and Crop Science: Selected Readings. Madison: Soil Science Society of America, 161-186.

Cromack H.T.H, Smith J.M. 1998. Calendula officinalis– production potential and crop agronomy in Southern England. Industrial CropsandProducts 7, 223-229.

Ekinci M, Turan M,  Yildirim E,  Guneu A,  Kotan R, Dursun A. 2014. Effect of plant growth promoting rhizobacteria on growth, nutrient, organic acid, amino acid and hormone content of cauliflower (Brassica oleracea L. var. botrytis) transplants. ActaScientiarum Polonorum Hortorum Cultus 13(6), 71-85.

10. El-Shabrawi H, Bakry B.A, Mohamed A.A, Abou-El-Lail M. 2015. Humic and oxalic acid stimulates grain yield and induces accumulation of plastidial carbohydrate metabolism enzymes in wheat grown under sandy soil conditions. Agricultural Sciences 6, 175-185.

11. Fallah S, Rostaei M, Lorigooini Z, Abbasi Surki A. 2018. Chemical compositions of essential oil and antioxidant activity of dragonhead (Dracocephalum moldavica) in sole crop and dragonhead soybean (Glycine max) intercropping system under organic manure and chemical fertilizers Industrial Crops & Products 115, 158-165.

12. Garcia-Seco D, Zhang Y. Gutierrez-Mañero F.J, Martin C, Ramos-Solano B. 2015. Application of Pseudomonas fluorescens to Blackberry under Field Conditions Improves Fruit Quality by Modifying Flavonoid Metabolism. PLoS ONE 10(11).

13. Gholami H, Ghani  A, Raouf Fard F, Saharkhiz M.J, Hazrati H. 2018a. Changes in photosynthetic pigments and uptake of some soil elements by chicory supplied with organic fertilizers. Acta Ecologica Sinica, In press.

14. Gholami H, Saharkhiz M.J, Raouf Fard F, Ghani A, Nadaf F. 2018b. Humic acid and vermicompost increased bioactive components, antioxidant activity and herb yield of Chicory (Cichorium intybus L.). Biocatalysis and Agricultural Biotechnology 14, 286-292.

15. Guinot, P,GargadenneA,ValetteG,Fruchier, A, AndaryC. 2008. Primary flavonoids in Marigold dye: Extraction, structure and involvement in the dyeing process. PhytochemicalAnalysis 19, 46-51.

16. Gutiérrez-MiceliF.A, García-GómezR.C,Oliva-LlavenM.A,Montes-MolinaJ.A,DendoovenL. 2017. Vermicomposting leachate as liquid fertilizer for the cultivation of sugarcane (Saccharum sp.). Journal of Plant Nutrition 40(1), 40-49.

17. He Y, Wu Z, Tu L, Shan C. 2017. Effect of encapsulated pseudomonas putida Rs-198 strain on alleviating salt stress of cotton. Journal of Plant Nutrition 40(8), 1180-1189.

18. Hosseinzadeh S.R, Amiri H, Ismaili A. 2018. Evaluation of photosynthesis, physiological, and biochemical responses of chickpea (Cicer arietinum L. cv. Pirouz) under water deficit stress and use of vermicompost fertilizer. Journal of Integrative Agriculture 17(11), 2426-2437.

19. Hwang M, Ederer G.M. 1975. Rapid hippurate hydrolysis method for presumptive identification of group streptococci. Journal of Clinical Microbiology1, 114-115.

    20.  Ipek M,Pirlak L,EsitkenA,Dönmez M.F,Turan M,Sahin F. 2014. Plant Growth-Promoting Rhizobacteria (Pgpr) Increase Yield, Growth And Nutrition Of Strawberry Under High-Calcareous Soil Conditions. Journal of Plant Nutrition 37(7), 990-1001.

    21. KhanA,Khan M.Z,Hussain F,Akhtar M.E,Gurmani A.R,Khan S. 2013. Effect of humic acid on the growth, yield, nutrient composition, photosynthetic pigment and total sugar contents of peas (pisum sativum L.). Journal of the Chemical Society of Pakistan 35(1), 206-211.

    22. Kochert G. 1978. Carbohydrate determination by the phenol-sulfuric acid method. In Hellebust J.A, Craigie J.S. Handbook of phycological methods: Physiological and Biochemical Methods. Cambridge University Press: Cambridge, 95-97. 

    23. Lichtenthaler H.K, Wellburn A.R. 1983. Deter-minations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions 11, 591-592.

    24. Meda A, Lamicn C.E, Romito M, Millogo J, Nacoulma O.G. 2005. Determination of the total phenolic, flavonoid and proline contents in Burkina fasan honey, as well as their radical scavenging activity. Food Chemistry 91, 571-577.

    25. Miliauskas G, Venskutonis P.R. Beek T.A. 2004. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry 85, 231-237.

    26. Mita S, Murano N, Akaike M, Nakamura K. 1997. Mutants of Arabidopsis thaliana with pleiotropic effects on the expression of the gene for beta-amylase and on the accumulation of anthocyanin that are inducible by sugars. The Plant Journal 11, 841-851.

      27.  Moreno-Reséndez A, Meza-Morales H, Rodríguez-Dimas N, Reyes-Carrillo J.L. 2010. Development of muskmelon with different mixtures of vermicompost: sand under greenhouse. Journal of Plant Nutrition 33(11), 1672-1680.

      28. Naderi D, Fallahzade J. 2017. Investigation of the potential use of recycling spent mushroom compost as Marigold (Calendula officinalis) bedding medium. Journal of Plant Nutrition 40(19), 2662-2668.

      29. Omara A, Hauka F, Afify A, Nour El-Din M, Kassem M. 2017. The role of some PGPR strains to biocontrol Rhizoctonia solani in soybean and enhancement the growth dynamics and seed yield. Environment,Biodiversityand Soil Security 1, 47-59.

      30. Onofrei V, Teliban G.C, Burducea M, Lobiuc A, Sandu C.B, Tocai M, Robu T. 2017. Organic foliar fertilization increases polyphenol content of Calendula ocinalis L. Industrial Crops & Products 109, 509-513.

      31. Parandian F, Samavat S. 2012. Effects of Fulvic and Humic acid on Anthocyanin, soluble Sugar, ∝ -Amylase Enzyme and some micronurient elements in Lilium. International Research Journal of Applied and Basic Sciences 3(5), 924-929.

      32. Porra R.J. 2002. The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. Photosynthesis Research 73(1-3), 149-56.

      33. Salehi A, Tasdighi H, Gholamhoseini M. 2016. Evaluation of proline, chlorophyll, soluble sugar content and uptake of nutrients in the German chamomile (Matricaria chamomilla L.) under drought stress and organic fertilizer treatments. Asian Pacific Journal of Tropical Biomedicine 12(10), 886-891.

      34. Schiavon M, Pizzeghello D,  Muscolo A, Vaccaro S,  Francioso O, Nardi S. 2010. High molecular size humic substances enhance phenylpropanoid metabolism in maize (Zea mays L.). JournalofChemical Ecology 36(6), 662-9.

      35. Simsek-Ersahin Y. 2011. The use of vermicompost products to control plant diseases and pests. In A. Karaca, Biology of earthworms: Soil Biology. Berlin Heidelberg: Springer-Verlag, 191-213.

      36. Song X, Liu M, Wu D, Griffiths B.S, Jiao J, Li H, Hu F. 2015. Interaction matters: Synergy between vermicompost and PGPR agents improves soil quality, crop quality and crop yield in the field. Applied Soil Ecology 89, 25-34.

      37. Sousa C, Pereira D.M, Pereira J.A, Bento A, Rodrigues M.A, Garcı´a D.S, Valenta˜ o P, Lopes G,  Ferreres F, Seabra R.M, Andrade P.B. 2008. Multivariate analysis of tronchuda cabbage (Brassica oleracea L. var. costata DC) phenolics: influence of fertilizers. Journal of Agricultural and Food Chemistry 56, 2231-2239.

      38. Tan K.H. 2003. Humic Matter in Soil and Environment: Principles and Controversies. New York: Marcel Dekker.

      39. Theunissen J, Ndakidemi P.A, Laubscher C.P. 2010. Potential of vermicompost produced from plant waste on the growth and nutrient status in vegetable production. International Journal of the Physical Sciences 5(13), 1964-1973.

      40. Tlili N, Elguizani T, Nasri N, Khaldi A, Triki S. 2011. Protein, lipid, aliphatic and triterpenic alcohols content ofcaper seedsCapparisspinosa”. Journal of the American OilChemists' Society88, 265-270.

      41. Trinh C.S, Jeong C.Y, Lee W.J, Truong H.A, Chung N, Han J, Hong S.W, Lee H. 2018. Paenibacillus pabuli strain P7S promotes plant growth and induces anthocyanin accumulation in Arabidopsis thaliana. Plant Physiology and Biochemistry 129, 264-272.

      42. Uma B, Malathi M. 2009. Vermicompost as a soil supplement to improve growth and yield of Amaranthus species. Research Journal of Agriculture and Biological Sciences 5(6), 1054-1060.