Document Type : Original Article

Authors

Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Background: Alfalfa (Medicago sativa L.) is an important source of phytoestrogens. The abundance of alkaloids, phenols, flavonoids, and isoflavonoids has made this plant a rich source of these plant estrogens. Cultivation of alfalfa as a rich source of phytoestrogens for medicinal purposes has provided opportunities for alternative use of this forage.
Methods: The study was carried out as a hydroponic culture in a perlite-cocopeat compacted bed with three replications. Roots and shoots of alfalfa plants were sampled separately in two stages (the 30th and 60th days after sowing). Plant samples were extracted with methanol solvent, and total phenols, flavonoids, and isoflavonoids contents were measured by spectrophotometric colorimetric method.
Results: The data analysis showed a significant effect of plant organ and harvest time on the contents of total phenols, flavonoids, and isoflavonoids (P≤0.05). The maximum accumulation of these compounds was in the plant shoots, and with the increase of the harvest time, the content of these phytoestrogens increased. Spearman's correlation analysis showed the different effects of the plant organ on the correlation level of the mentioned metabolites, so the flavonoids of the roots and shoots showed the most positive correlation, while isoflavonoids did not show a significant correlation (P≤0.05).
Conclusions: The presence of the maximum contents of total phenols, flavonoids, and isoflavonoids in the shoots of alfalfa can be concluded that the distribution of secondary metabolites in plants, the same as the primary metabolites, is mainly dependent on the plant organ and tissue. Furthermore, the maximum content of these metabolites in the vegetative stage of alfalfa is due to the transition from the vegetative stage to the reproductive stage in these plants. Therefore, the late vegetative stage is the best phenological stage and the most suitable harvest time for medicinal applications.

Graphical Abstract

The Correlation of Some Secondary Metabolites of Alfalfa (Medicago sativa L.) with Plant Organ and Harvest Time

Keywords

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  1. Bora KS, Sharma A. Phytochemical and pharmacological potential of Medicago sativa: A review, Pharmaceutical biology; 2011 Feb 1; 49(2):211-20. [Crossref], [Google Scholar], [Publisher]
  2. Gholami A, De Geyter N, Pollier J, Goormachtig S, Goossens A. Natural product biosynthesis in Medicago species, Natural Product Reports; 2014; 31(3):356-80. [Crossref], [Google Scholar], [Publisher]
  3. Shailajan S, Sayed N, Menon S, Singh A, Mhatre M. A validated RP-HPLC method for quantitation of trigonelline from herbal formulations containing Trigonella foenum-graecum (L.) seeds, Pharmaceutical Methods; 2011 Jul 1; 2(3):157-60. [Crossref], [Google Scholar], [Publisher]
  4. Ludwig IA, Clifford MN, Lean ME, Ashihara H, Crozier A. Coffee: biochemistry and potential impact on health, Food & Function; 2014; 5(8):1695-717. [Crossref], [Google Scholar], [Publisher]
  5. Preedy VR, editor, Coffee in health and disease prevention; Academic Press; 2014 Nov 12. [Crossref], [Google Scholar], [Publisher]
  6. Ashihara H, Ludwig IA, Katahira R, Yokota T, Fujimura T, Crozier A. Trigonelline and related nicotinic acid metabolites: occurrence, biosynthesis, taxonomic considerations, and their roles in planta and in human health, Phytochemistry Reviews; 2015 Oct; 14:765-98. [Crossref], [Google Scholar], [Publisher]
  7. Soto-Zarazúa MG, Rodrigues F, Pimentel FB, Bah MM, Oliveira MB. The isoflavone content of two new alfalfa-derived products for instant beverage preparation, Food & function; 2016; 7(1):364-71. [Crossref], [Google Scholar], [Publisher]
  8. Moore KJ, Collins M, Nelson CJ, Redfearn DD, editors. Forages, Volume 2: The science of grassland agriculture; John Wiley & Sons; 2020 Aug 24. [Crossref], [Google Scholar], [Publisher]
  9. Katoch R. Nutritional Quality Estimation of Forages. InNutritional Quality Management of Forages in the Himalayan Region 2022 Apr 13 (pp. 225-278). Singapore: Springer Singapore. [Crossref], [Google Scholar], [Publisher]
  10. Yildiz F, editor. Phytoestrogens in functional foods, CRC Press; 2019 Jul 17. [Crossref], [Google Scholar], [Publisher]
  11. César ID, Braga FC, Vianna-Soares CD, Nunan ED, Pianetti GA, Moreira-Campos LM. Quantitation of genistein and genistin in soy dry extracts by UV-Visible spectrophotometric method, Química Nova; 2008; 31(8):1933-6. [Crossref], [Google Scholar], [Publisher]
  12. Saunders JA, Matthews BF, Romeo J. Regulation of phytochemicals by molecular techniques, Elsevier; 2001 Jul 23. [Google Scholar], [Publisher]
  13. Du H, Huang Y, Tang Y. Genetic and metabolic engineering of isoflavonoid biosynthesis, Applied Microbiology and Biotechnology; 2010 May; 86:1293-312. [Crossref], [Google Scholar], [Publisher]
  14. Seguin P, Zheng W. Phytoestrogen content of alfalfa cultivars grown in eastern Canada, Journal of the Science of Food and Agriculture; 2006 Apr 15; 86(5):765-71. [Crossref], [Google Scholar], [Publisher]
  15. Atumo T, Jones CS. Varietal differences in yield and nutritional quality of alfalfa (Medicago sativa) accessions during 20 months after planting in Ethiopia, Tropical Grasslands-Forrajes Tropicales; 2021 Jan 30; 9(1):89-96. [Crossref], [Google Scholar], [Publisher]
  16. Hoagland DR, Arnon DI. The water-culture method for growing plants without soil, Circular, California agricultural experiment station; 1950; 347(2nd edit). [Crossref], [Google Scholar], [Publisher]
  17. Ismail HI, Chan KW, Mariod AA, Ismail M. Phenolic content and antioxidant activity of cantaloupe (Cucumis melo) methanolic extracts, Food chemistry; 2010 Mar 15; 119(2):643-7. [Crossref], [Google Scholar], [Publisher]
  18. Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colometric methods, Journal of food and drug analysis; 2002; 10(3):3. [Crossref], [Google Scholar], [Publisher]
  19. Turgut BA, Bezirganoglu İ. Callus Induction and Bioactive Compounds Production from Various Cultivars of Medicago sativa L.(alfalfa), Journal of the Institute of Science and Technology; 2023 Sep 1; 13(3):1625-32. [Crossref], [Google Scholar], [Publisher]
  20. Butkutė B, Padarauskas A, Cesevičienė J, Taujenis L, Norkevičienė E. Phytochemical composition of temperate perennial legumes, Crop and Pasture Science; 2018 Aug 31; 69(10):1020-30. [Crossref], [Google Scholar], [Publisher]
  21. Krakowska A, Rafińska K, Walczak J, Kowalkowski T, Buszewski B. Comparison of various extraction techniques of Medicago sativa: yield, antioxidant activity, and content of phytochemical constituents, Journal of AOAC International; 2017 Nov 1; 100(6):1681-93. [Crossref], [Google Scholar], [Publisher]
  22. RUGNA AZ, GURNI AA, WAGNER ML. Phenological variations of polyphenols in Smilax campestris (Smilacaceae), Turkish Journal of Botany; 2013; 37(2):350-4. [Crossref], [Google Scholar], [Publisher]
  23. Lee J, Jung J, Son SH, Kim HB, Noh YH, Min SR, Park KH, Kim DS, Park SU, Lee HS, Kim CY. Profiling of the major phenolic compounds and their biosynthesis genes in Sophora flavescens aiton, The Scientific World Journal; 2018 Jan 1; 2018. [Crossref], [Google Scholar], [Publisher]
  24. Oliveira I, Valentão P, Lopes R, Andrade PB, Bento A, Pereira JA. Phytochemical characterization and radical scavenging activity of Portulaca oleraceae L. leaves and stems, Microchemical Journal; 2009 Jul 1; 92(2):129-34. [Crossref], [Google Scholar], [Publisher]
  25. Petropoulos SA, Fernandes Â, Dias MI, Vasilakoglou IB, Petrotos K, Barros L, Ferreira IC. Nutritional value, chemical composition and cytotoxic properties of common purslane (Portulaca oleracea L.) in relation to harvesting stage and plant part, Antioxidants; 2019 Aug 8; 8(8):293. [Crossref], [Google Scholar], [Publisher]
  26. Páramo L, Feregrino-Pérez AA, Vega-González M, Escobar-Alarcón L, Esquivel K. Medicago sativa L. Plant Response against Possible Eustressors (Fe, Ag, Cu)-TiO2: Evaluation of Physiological Parameters, Total Phenol Content, and Flavonoid Quantification, Plants; 2023 Feb 2; 12(3):659. [Crossref], [Google Scholar], [Publisher]
  27. Seguin P, Zheng W, Souleimanov A. Alfalfa phytoestrogen content: Impact of plant maturity and herbage components, Journal of Agronomy and Crop Science; 2004 Jun; 190(3):211-7. [Crossref], [Google Scholar], [Publisher]
  28. Caunii A, Pribac G, Grozea I, Gaitin D, Samfira I. Design of optimal solvent for extraction of bio–active ingredients from six varieties of Medicago sativa, Chemistry Central Journal; 2012 Dec; 6(1):1-8. [Crossref], [Google Scholar], [Publisher]
  29. Ayan AK, Yanar P, Cirak C, Bilgener M. Morphogenetic and diurnal variation of total phenols in some Hypericum species from Turkey during their phenological cycles, Bangladesh Journal of Botany; 2007; 36(1):39-46. [Crossref], [Google Scholar]
  30. ÇIRAK C, Radusiene J, Ivanauskas L, Jakstas V, ÇAMAŞ N. Phenological changes in the chemical content of wild and greenhouse-grown Hypericum pruinatum: flavonoids, Turkish Journal of Agriculture and Forestry; 2014; 38(3):362-70. [Crossref], [Google Scholar], [Publisher]
  31. Saeb K, Gholamrezaee S, Asadi MA. Variation of antioxidant activity of Melissa officinalis leaves extracts during the different stages of plant growth, Biomedical and Pharmacology Journal; 2011; 4(2):237-43. [Crossref], [Google Scholar], [Publisher]
  32. Nurzyńska-Wierdak R. Phenolic Compounds from New Natural Sources—Plant Genotype and Ontogenetic Variation, Molecules; 2023 Feb 11; 28(4):1731. [Crossref], [Google Scholar], [Publisher]
  33. Sharma A, Shahzad B, Rehman A, Bhardwaj R, Landi M, Zheng B. Response of phenylpropanoid pathway and the role of polyphenols in plants under abiotic stress, Molecules; 2019 Jul 4; 24(13):2452. [Crossref], [Google Scholar], [Publisher]
  34. Soufan W, Okla MK, Salamatullah A, Hayat K, Abdel-Maksoud MA, Al-Amri SS. Seasonal variation in yield, nutritive value, and antioxidant capacity of leaves of alfalfa plants grown in arid climate of Saudi Arabia, Chilean journal of agricultural research; 2021 Jun; 81(2):182-90. [Crossref], [Google Scholar], [Publisher]
  35. Rodrigues F, Almeida I, Sarmento B, Amaral MH, Oliveira MB. Study of the isoflavone content of different extracts of Medicago spp. as potential active ingredient, Industrial Crops and Products; 2014 Jun 1; 57:110-5. [Crossref], [Google Scholar], [Publisher]
  36. Chouaieb H, Ayadi I, Zouari S, Fakhfakh N, Zaidi S, Zouari N. Effect of phenological stage and geographical location on antioxidant activities of tunisian horehound: Marrubium vulgare L.(Lamiaceae), Journal of Biologically Active Products from Nature; 2012 Jan 1; 2(4):232-8. [Crossref], [Google Scholar], [Publisher]
  37. Li Y, Zidorn C. Seasonal variations of natural products in European herbs, Phytochemistry Reviews; 2022 Oct; 21(5):1549-75. [Crossref], [Google Scholar], [Publisher]
  38. Szakiel A, Pączkowski C, Henry M. Influence of environmental abiotic factors on the content of saponins in plants, Phytochemistry Reviews; 2011 Dec; 10:471-91. [Crossref], [Google Scholar], [Publisher]
  39.  Yang L, Wen KS, Ruan X, Zhao YX, Wei F, Wang Q. Response of plant secondary metabolites to environmental factors, Molecules; 2018 Mar 27; 23(4):762. [Crossref], [Google Scholar], [Publisher