The effect of different concentrations of chitosan on the production of phenolic acids in cell culture of Lactuca undulate Ledeb.

Document Type : Research Paper

Authors

1 Ph.D. Student, Department of Biology, Faculty of Science, Golestan University, Gorgan, Iran.

2 Associated Professor, Department of Biology, Faculty of Science, Golestan University, Gorgan, Iran.

3 Assistant Professor, Department of Biology, Faculty of Science, Golestan University, Gorgan, Iran.

Abstract

Objective
One of the most important methods for increasing the production of secondary metabolites is the use of elicitors in plant cell culture. A biopolymer made from D-glucosamine units, chitosan can be found in the cell walls of fungi and the exoskeletons of arthropods. Plants respond to chitosan treatment by generating defense responses, increasing antioxidant enzyme activity, accumulating phenolic compounds, and releasing flavonoids. In the current study, chitosan was used as an elicitor to induce the production of phenolic acids in Lactuca undulata cells suspended in a liquid medium.
 
Materials and methods
First, cell suspension culture was prepared from 45 day old callus derived from leaf explants of Lactuca undulata on ½ MS medium supplemented with 0.1 and 1 mg/L 2,4-D and Kin. The effect of different concentrations of chitosan (0, 50, 100, and 200 mg/L) on cell suspension was evaluated during 24, 48, and 72 hours. After harvesting samples, the percentage of cell viability, total phenols and flavonoids, chicoric acid, chlorogenic acid, and caffeic acid contents, as well as phenylalanine-ammonia lyase (PAL) activity and lipid peroxidation were measured.
 
Results
In the present study, we found that the concentration and duration of chitosan treatment affect the production of phenolic compounds (including chicoric acid) in the Lactuca undulata cell suspension culture. After 24 hours of treatment with 50 mg/L chitosan, chicoric acid levels had increased 2.8-fold compared to the control. After 24 and 48 hours of treatment with 200 mg/L chitosan, the highest levels of chlorogenic and caffeic acids were observed. Furthermore, the present study found the chitosan treatment resulted in an increase in the amount of phenol and total flavonoids as well as an increase in PAL enzyme activity. Chitosan induces lipid peroxidation and reduces cell viability in high concentrations, indicating a negative effect.
 
Conclusions
The present study found that low concentrations of chitosan could induce chicoric acid production in Lactuca undulata cell suspension cultures, which can be utilized in the pharmaceutical industry as a new method in the production of chicoric acid and its derivatives.

Keywords

References

 
Ahmad W, Zahir A, Nadeem M et al. (2018) Enhanced production of lignans and neolignans in chitosan-treated flax (Linum usitatissimum L.) cell cultures, Process Biochemistry. https://doi.org/10.1016/j.procbio.2018.12.025.
 Cabrera JC, Messiaen J, Cambier P, Van Cutsem P (2006) Size, acetylation and concentration of chitooligosaccharide elicitors determine the switch from defence involving PAL activation to cell death and water peroxide production in Arabidopsis cell suspensions. Physiol Plant 127, 44-56.
Chakrabortya M, Karunb A, Mitra A (2009) Accumulation of phenylpropanoid derivatives in chitosan-induced cell suspension culture of Cocos nucifera. J Plant Physiol 166, 63-71.
Chang C, Yang M, Wen H, Chern J (2002) Estimation of total flavonoid content in Propolis by Two complementary colorimetric Methods. J Food and Druge Analysis 10, 178-182.
Fernández-Bautista N, Domínguez-Núñez JA, Castellano MM, Berrocal-Lobo M (2016) Plant Tissue Trypan Blue Staining During Phytopathogen Infection. Bio-Protocol 6, e2078
Ferri M, Tassoni A (2011) Chitosan as elicitor of health beneficial secondary metabolites in in vitro plant cell cultures. In R. G. Mackay & J. M. Tait (Eds.), Handbook of chitosan research and applications 389-414. Nova Science Publishers.
Hall RD (2000) Plant Cell Culture initiation. Practical tips. Mol Biotechnol 16, 161-173.
Hammerschmidt R (2014) Chlorogenic acid: A versatile defense compound. Physiol Mol Plant Pathol 88, e35.
Hudec J, Burdova M, Kobida L, et al. (2007) Antioxidant Capacity Changes and Phenolic Profile of Echinacea purpurea, Nettle (Urtica dioica L.) and Dandelion (Taraxacum officinale) after Application of Polyamine and Phenolic Biosynthesis Regulators. J Agric Food Chem 55, 5689−5696.
Iriti M, Giulia C, Sara V, et al. (2010) Chitosan-induced ethylene-independent resistance does not reduce crop yield in bean. Biol Control 54, 241-247.
Kamalipourazad M, Sharifi M, Zare Maivan H, Behmanesh M et al. (2016) Induction of aromatic amino acids and phenylpropanoid compounds in Scrophularia striata Boiss. Cell culture in response to chitosan-induced oxidative stress. Plant Physiol Biochem107, 374-384.
Kazi GAS, Yamanaka T, Osamu Y (2019) Chitosan coating an Efficient Approach to Improve the Substrate Surface for In Vitro Culture System. J Electrochem Soc 166, B3025
Khan W, Prithiviraj B, Smith DL (2003) Chitosan and chitin oligomers increase phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities in soybean leaves. J Plant Physiol 160, 859-863.
Landi L, Feliziani E, Romanazzi G (2014) Expression of Defense Genes in Strawberry Fruits Treated with Different Resistance Inducers. J Agric Food Chem 62, 3047−3056.
Lee J, Scagel CF (2013) Chicoric acid: chemistry, distribution, and production. Front Chem 1, 117-123.
Luo XB, Chen B, Yao S Z, Zeng JG (2003) Simultaneous analysis of caffeic acid derivatives & alkamides in roots and extracts of Echinacea purpurea by HPLC-photodiode array detection-electrospray mass spectrometry. J Chromatogr 986, 73-81.
Meda A, Euloge C, Romito M, et al. (2005) Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity. Food Chem 91, 571-577.
Mejía-Teniente L, Dalia Duran-Flores F, Chapa-Oliver AM, et al. (2013) Oxidative and molecular responses in Capsicum annuum L. after hydrogen peroxide, salicylic acid and chitosan foliar applications. Int J Mol Sci 14,10178-10196.
Mhlongo MI, Piater LA, Madala NE, et al. (2016) Phenylpropanoid Defences in Nicotiana tabacum Cells: Overlapping Metabolomes Indicate Common Aspects to Priming Responses Induced by Lipopolysaccharides, Chitosan and Flagellin-22. PLoS One 11, e0151350.
Mofid Bojnoordi M, Aghdasi M, Fatemi M (2020) An investigation on phytochemical components and antioxidant activity of Luctuca undulate in 5 natural habitats of Iran. Medic Plant 75, 65-75.
Nair VD, Panneerselvam R, Gopi R, Hong-bo S (2013) Elicitation of pharmacologically active phenolic compounds from Rauvolfia serpentina Benth. Ex. Kurtz. Ind Crops Prod 45, 406-415.
Nuissier G, Rezzonico B, Grignon-Dubois M 2010. Chicoric acid from Syringodium filiforme. Food Chem 120, 783-788.
Omidbaigi R (2002) Study of cultivation and adaptability of purple coneflower (Echinaceae purpurea) in the North of Tehran. JWSS-Isfahan University of Technology, 6 231-241.
Prochazkova D, Sairam RK, Srivastava GC, Singh DV (2001) Oxidative stress and antioxidant activity as the basis of senescence in maize leaves. Plant Sci 161, 765-771.
Ramezannezhad R (2018) An investigation on antioxidative activity and Caffeic acid derivatives production in some Iranian native species of Asteraceae family under nature and tissue culture condition. PhD thesis, Golestan University. pp:124-139.
Ramezannezhad R, Aghdasi M, Fatemi M (2019) An investigation on cichoric acid content and antioxidant activity in some Iranian native species compared to Echinacea purpurea L. in different developmental stages. Iran J Medic Aromatic Plant 34, 909-923.
Singh S (2014) A review on possible elicitor molecules of cyanobacteria: Their role in improving plant growth and providing tolerance against biotic or abiotic stress. J Appl Microbiol 117, 1221-1244.
Singh S (2016) Enhancing phytochemical levels, enzymatic and antioxidant activity of spinach leaves by chitosan treatment and an insight into the metabolic pathway using DART-MS technique. Food Chem 199, 176-184.
Sircar D, Mitra A (2009) Accumulation of p-hydroxybenzoic acid in hairy roots of Daucus carota 2: Confirming biosynthetic steps through feeding of inhibitors and precursors. J Plant Physiol 166, 1370-1380.
Sykłowska-Baranek k, Pietrosiuk A, Naliwajski MR, et al. (2012) Effect of L-phenylalanine on PAL activity and production of naphthoquinone pigments in suspension cultures of Arnebia euchroma (Royle) Johnst. In Vitro Cell Dev Biol-Plant 48, 555-564.
Valletta A, De Angelis G, Badiali C, et al. (2016) Acetic acid acts as an elicitor exerting a chitosan-like effect on xanthone biosynthesis in Hypericum perforatum L. root cultures. Plant Cell Rep 35, 1009-1020.
Vasconsuelo A, Picotto G, Giuletti AM, Boland R (2006). Involvement of G‐proteins in chitosan‐induced Anthraquinone synthesis in Rubia tinctorum. Physiol Plantarum 128, 29-37.
Yin H, Frette XC, Christensen LP, Grevsen K (2012) Chitosan oligosaccharides promote the content of polyphenols in Greek oregano (Origanum vulgare ssp. hirtum). J Agric Food Chem 60, 136-143.
Zhang X, Li K, Xing R, et al. (2018) miRNA and mRNA Expression Profiles Reveal Insight into Chitosan-Mediated Regulation of Plant Growth. J Agric Food Chem 66, 3810-3822.
Zlotek U, Swieca M (2016) Elicitation effect of Saccharomyces cerevisiae yeast extract on main health-promoting compounds and antioxidant and anti-inflammatory potential of butter lettuce (Lactuca sativa L.). J Sci Food Agric 96, 2565-2572.
Zuppini A, Baldan B, Millioni R, Favaron F et al. (2003) Chitosan induces Ca Blackwell Publishing, Ltd. 2+-mediated programmed cell death in soybean cells. New Phytologist 161, 557-568
Agricultural Biotechnology Journal
Volume 14, Issue 3
September 2022
Pages 1-20

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