اثر ترکیبی کیتوزان و هورمون سالیسیلیک اسید بر صفات فیزیولویکی، بیوشیمیایی و بیان ژن‌های کلیدی مسیر بیوسنتز کارنوسیک‌اسید در گیاه رزماری (Rosmarinus officinalis L.)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه زیست‌شناسی، دانشگاه پیام نور، صندوق پستی ۴۶۹۷-۱۹۳۹۵، تهران، ایران.

2 استادیار، گروه زیست‌شناسی، دانشگاه پیام نور، صندوق پستی ۴۶۹۷-۱۹۳۹۵، تهران، ایران.

3 دانشیار، گروه زیست‌شناسی، دانشگاه پیام نور، صندوق پستی ۴۶۹۷-۱۹۳۹۵، تهران، ایران.

4 استاد، گروه زیست‌شناسی، دانشگاه پیام نور، صندوق پستی ۴۶۹۷ـ۱۹۳۹۵، تهران، ایران.

10.22103/jab.2026.25770.1752

چکیده

هدف: رزماری (Rosmarinus officinalis L) یکی از گیاهان دارویی و معطر مهم در صنعت دارو و غذا به حساب می­آید. در میان ترپن­های رزمای، کارنوسیک ­اسید به دلیل خواص درمانی نظیر آنتی­اکسیدانی، ضدمیکروبی و ضدتوموری اهمیت بسزائی یافته است. درنتیجه، توجه محققان به کاربرد الیسیتورها برای تقویت رشد و افزایش اسانس رزماری جلب شده است. با توجه به اهمیت دارویی رزماری و پتانسیل کیتوزان و سالیسیلیک ­اسید در تقویت صفات گیاهی، این پژوهش با هدف بررسی اثر این الیسیتورها بر صفات رزماری و بیان ژن­های کلیدی مسیر کارنوسیک­ اسید (CPS1 و KSL1) انجام شد.
مواد و روش‌ها: این پژوهش در قالب آزمایش فاکتوریل با طرح کاملا تصادفی با سه تکرار انجام شد. بعد از گذشت سه ماه از کشت نشاها، کیتوزان (Ch) با غلظت 10، 50 و 100 میلی­گرم بر لیتر و سالیسیلیک ­اسید (SA) با غلظت 5، 10 و 20 میکرو‌مولار به طور انفرادی و در ترکیب باهم در سه نوبت با فاصله سه هفته بر روی اندام هوایی رزماری محلول­پاشی شدند. بعد از گذشت یک هفته از آخرین محلول­پاشی، سنجش صفات فیزیولویکی، بیوشیمیایی و بیان ژن­های کلیدی بیوسنتز کارنوسیک اسید انجام شد.
نتایج: ترکیب­های تیماری، اثر هم­افزائی بر صفات رزماری داشتند. به عنوان موثرترین تیمار، تیمارهایSA 10 µM + Ch 100 mg/L  و SA 20 µM + Ch 50 mg/L به ترتیب سبب کاهش معنی­دار حدود 44 و 45% مالون­دی­آلدئید و افزایش معنی­دار 2 و 5/2 برابری فنل، 8/2 و 9/2 برابری قندها، 6/2 و 2 برابری فعالیت کاتالاز، 8/2 و 5/2 برابری فعالیت آسکوربات­پراکسیداز، 2 و 3/2 برابری فعالیت پلی­فنل­اکسیداز، 3 و 9/1 برابری کلروفیل، 2 و 2/2 برابری کارتنوئید، 3/2 و 6/2 برابری اسانس، 3 و 6/3 برابری بیان CPS1، و 4/3 و 6/3 برابری بیان KSL1 شدند.
نتیجه‌گیری: تیمارهای ترکیبی سبب کاهش معنی­دار اکسیداسیون لپیدی و افزایش رنگدانه­ها، سامانه آنتی­اکسیدانت، اسانس و بیان ژن­های کلیدی مسیر کارنوسیک ­اسید می­شوند.

کلیدواژه‌ها

عنوان مقاله [English]

Combined effect of chitosan and salicylic acid hormone on physiological and biochemical traits and expression of key genes of carnosic acid biosynthesis pathway in rosemary (Rosmarinus officinalis L.)

نویسندگان [English]

  • Morteza Najafpour Mohammadabady 1
  • Enayatollah Yazdanpanah 2
  • Hamid Sobhanian 3
  • Gholamreza Bakshi Khaniki 4
1 Ph.D. Student, Department of Biology, Payame Noor University (PNU), P.O. Box 19395-4697, Tehran, Iran.
2 Assistant Professor, Department of Biology, Payame Noor University (PNU), P.O. Box 19395-4697, Tehran, Iran.
3 Associate Professor, Department of Biology, Payame Noor University (PNU), P.O. Box 19395-4697, Tehran, Iran.
4 Professor, Department of Biology, Payame Noor University (PNU), P.O. Box 19395-4697, Tehran, Iran.
چکیده [English]

Objective
Rosemary (Rosmarinus officinalis L.), as one of the important medicinal and aromatic plants in the pharmaceutical and food industries. Among the terpenes of rosemary, carnosic acid has gained significant importance due to its therapeutic properties such as antioxidant, antimicrobial, and antitumor effects. Consequently, researchers' attention has been drawn to the application of elicitors to enhance the growth and increase the essential oil of rosemary. Given the medicinal importance of rosemary and the potential of chitosan and salicylic acid in enhancing plant traits, this research was conducted with the aim of investigating the effect of these elicitors on rosemary traits and the expression of key genes in the carnosic acid pathway (KSLI and CPSI).
Materials and methods
This research was carried out as a factorial experiment based on a completely randomized design with three replications. Three months after transplanting the seedlings, chitosan (Ch) at concentrations of 10, 50, and 100 mg/L and salicylic acid (SA) at concentrations of 5, 10, and 20 µM were applied individually and in combination, in three stages at three-week intervals, as foliar sprays on the aerial parts of rosemary. Physiological and biochemical traits and expression of key genes of carnosic acid biosynthesis pathway were measured one week after the last spraying.
Results
The treatment combinations had synergistic effects on rosemary traits. As the most effective treatments, SA 10 µM + Ch 100 mg/L and SA 20 µM + Ch 50 mg/L treatments led to a 44 and 45% significant decrease in MDA, 2 and 2.5-fold significant increase in phenol, 2.8 and 2.9-fold increase in sugars, 2.6 and 2-fold increase in catalase activity, 2.8 and 2.5-fold increase in SOD activity, 2 and 2.3-fold increase in polyphenol oxidase activity, 3 and 1.9-fold increase in chlorophyll content, 2 and 2.2-fold increase in carotenoid content, 2.3 and 2.6-fold increase in essential oil content, 3 and 3.6-fold increase in CPSI gene expression, 3.4 and 3.6-fold increase in KSLI gene expression, respectively.
Conclusions
Combined treatments significantly reduce lipid peroxidation and increase pigments, the antioxidant system, essential oil, and the expression of key genes in the carnosic acid pathway.

کلیدواژه‌ها [English]

  • Antioxidant
  • Carnosic acid
  • Elicitor
  • Rosemary

مراجع

Alizadeh, A., Moghaddam, M., Asgharzade, A., & Sourestani, M. M. (2020). Phytochemical and physiological response of Satureja hortensis L. to different irrigation regimes and chitosan application. Industrial Crops and Products, 158, 112990.  https://doi.org/10.1016/j.indcrop.2020.112990
Amirteymoori, E., Khezri, A., Dayani, O., Mohammadabadi, M., Khorasani, S., Mousaie, A., & Kazemi-Bonchenari, M. (2021). Effects of linseed processing method (ground versus extruded) and dietary crude protein content on performance, digestibility, ruminal fermentation pattern, and rumen protozoa population in growing lambs. Italian Journal of Animal Science, 20(1), 1506-1517. https://doi.org/10.1080/1828051X.2021.1984324
Angouti, F., Nourafcan, H., Saeidi Sar, S., Asadi, A., & Ebrahimi, R. (2022). The effect of different levels of chitosan and salicylic acid on morphological traits of the medicinal plant Galega (Galega officinalis). Journal of Crops Improvement, 24(4), 1341-1358. https://doi.org/10.22059/jci.2022.329753.2607
Armon, D. I. (1949). Copper enzymes in isolated chloroplasts. Plant Physiology, 24(1), 1-5.
Azadvari, H., Naeemi, M., Gholizadeh, A., & Nakhzari Moghaddam, A. (2019). Effect of salicylic acid on some physiological traits and yield of black cumin medicinal plant (Nigella sativa L.) under different humidity regimes. Journal of Crop Production, 12(3), 85-100. https://doi.org/10.22069/EJCP.2019.16031.2192
Bayrak, S., Öztürk, C., Demir, Y., Alım, Z., & Küfrevioglu, Ö. İ. (2020). Purification of polyphenol oxidase from potato and investigation of the inhibitory effects of phenolic acids on enzyme activity. Protein and Peptide Letters, 27(3), 187-192. https://doi.org/10.2174/0929866526666191002142301
Da Silva, E. A., Silva, V. N., De Alvarenga, A. A., & Bertolucci, S. K. (2021). Biostimulating effect of chitosan and acetic acid on the growth and profile of the essential oil of Mentha arvensis L. Industrial Crops and Products, 171, 113987. https://doi.org/10.1016/j.indcrop.2021.113987
Dhindsa, R. S., Plumb-Dhindsa, P., & Thorpe, T. A. (1981). Leaf senescence: Correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. Journal of Experimental Botany, 32(1), 93-101. https://doi.org/10.1093/jxb/32.1.93
Heath, R. L., & Packer, L. (1968). Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics, 125(1), 189-198. https://doi.org/10.1016/0003-9861(68)90654-1
Jahan, S., Anjali, K., Panwar, M., Mishra, R., Shankhdhar, S. C., & Shankhdhar, D. (2024). Integrative impacts of salicylic acid and water deficit stress on physiological processes of medicinal herb Bacopa monnieri (L.). Plant Physiology Reports, 29(1), 65-75. https://doi.org/10.1007/s40502-023-00770-4
Jami, S., Esmaeilzadeh Bahabadi, S., & Modarres, M. (2018). Effect of chitosan on micropropagation, secondary metabolites content and antioxidant activity of in vitro cultured Salvia leriifolia. Journal of Plant Research (Iranian Journal of Biology), 31(3), 568-578. https://doi.org/20.1001.1.23832592.1397.31.3.7.7
Khalafi, M., Pourbeyrami Hir, Y., Chamani, E., & Maleki Lajayer, H. (2022). The effect of chitosan on regeneration and secondary metabolite production of two species of lily flower. Journal of Plant Research, 35(2), 374-388. https://doi.org/20.1001.1.23832592.1401.35.2.9.5
Khezri, A., Shafabakhsh, H., Alizadeh, A., Mohammadabadi, M., & Shakeri, M. (2025). Effects of Encapsulated Mixtures of Plant Essential Oils and Organic Acids as an Alternative to Antibiotic Growth Promoters on Humoral Immune Response and Expression of Interleukin-4 and Interferon-Gamma Genes in Broilers. Journal of Poultry Sciences and Avian Diseases 3(3), 12-19. https://doi.org/10.61838/kman.jpsad.3.3.3  
Kumari, S., Singh, A. K., Kumar, A., Singh, K. P., & Bains, G. (2021). Evaluating the efficacy of chitosan and salicylic acid on photosynthetic pigments and antioxidant enzymes towards resistance of mango malformation. Scientia Horticulturae, 285, 110160. https://doi.org/10.1016/j.scienta.2021.110160
Li, K., Xing, R., Liu, S., & Li, P. (2020). Chitin and chitosan fragments responsible for plant elicitor and growth stimulator. Journal of Agricultural and Food Chemistry, 68(44), 12203-12211. https://doi.org/10.1021/acs.jafc.0c05316
Macedo, L. M., Santos, É. M. D., Militão, L., Tundisi, L. L., Ataide, J. A., Souto, E. B., & Mazzola, P. G. (2020). Rosemary (Rosmarinus officinalis L., syn Salvia rosmarinus Spenn.) and its topical applications: A review. Plants, 9(5), 651. https://doi.org/10.3390/plants9050651 137
Miclea, I., Suhani, A., Zahan, M., & Bunea, A. (2020). Effect of jasmonic acid and salicylic acid on growth and biochemical composition of in-vitro-propagated Lavandula angustifolia Mill. Agronomy, 10(11), 1722. https://doi.org/10.3390/agronomy10111722
Mohammadabadi, M., Afsharmanesh, M., Khezri, A., Kheyrodin, H., Babenko, O., Borshch, O. O., Kalashnyk, O., Nechyporenko, O., Afanasenko, V., Slynko, V., & Usenko, S. (2025). Effect of mealworm on GBP4L gene expression in the spleen tissue of Ross broiler chickens. Agricultural Biotechnology Journal, 17(2), 343-360. https://doi.org/10.22103/jab.2025.25277.1714
Mohammadabadi, M., Babenko, O., Borshch, O. O., Kalashnyk, O., Ievstafiieva, Y., & Buchkovska, V. (2024). Measurement of the relative expression pattern of the UCP2 gene in different tissues of the Raini Cashmere goat. Agricultural Biotechnology Journal, 16(3), 317-332. https://doi.org/10.22103/jab.2024.24337.1627
Mohammadabadi, M., Golkar, A., & Askari Hesni, M. (2023). The effect of fennel (Foeniculum vulgare) on insulin-like growth factor 1 gene expression in the rumen tissue of Kermani sheep. Agricultural Biotechnology Journal, 15(4), 239-256. https://doi.org/10.22103/jab.2023.22647.1530
Mohammadabadi, M., Shaban Jorjandy, D., Arabpoor Raghabadi, Z., Abareghi, F., Sasan, H. A., & Bordbar, F. (2022). The role of fennel on DLK1 gene expression in sheep heart tissue. Agricultural Biotechnology Journal, 14(2), 155-170. https://doi.org/10.22103/jab.2022.19402.1399
Momeni, M., Pirbalouti, A. G., Mousavi, A., & Badi, H. N. (2020). Effect of foliar applications of salicylic acid and chitosan on the essential oil of Thymbra spicata L. under different soil moisture conditions. Journal of Essential Oil Bearing Plants, 23(5), 1142-1153. https://doi.org/10.1080/0972060X.2020.1828178
Nakano, Y., & Asada, K. (1981). Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant and Cell Physiology, 22(5), 867-880. https://doi.org/10.1093/oxfordjournals.pcp.a076232
Naderi, S., Esmaeilzadeh Bahabadi, S., & Fakheri, B. (2015). The effect of chitosan on some physiological and biochemistry characterization in basil (Ocimum basilicum). Journal of Plant Process and Function, 4(12), 29-41. https://dor.isc.ac/dor/20.1001.1.23222727.1394.4.12.9.9
Peng, Y., Yang, J., Li, X., & Zhang, Y. (2021). Salicylic acid: Biosynthesis and signaling. Annual Review of Plant Biology, 72(1), 761-791. https://doi.org/10.1146/annurev-arplant-081320-092855
Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research, 29(9), e45. https://doi.org/10.1093/nar/29.9.e45
Poznanski, P., Shalmani, A., Bryla, M., & Orczyk, W. (2024). Salicylic acid mediates chitosan-induced immune responses and growth enhancement in barley. International Journal of Molecular Sciences, 25(24), 13244. https://doi.org/10.3390/ijms252413244
Rahmani, M., Kheiry, A., Sanikhani, M., Mortazavi, N., & Farzin, N. (2020). Effect of foliar application of chitosan on some physiological characteristics of Mentha spicata L. under deficit irrigation stress. Iranian Journal of Horticultural Science, 51(2), 479-492. https://doi.org/10.22059/ijhs.2019.271832.1565 1
Rayanoothala, P. S., Dweh, T. J., Mahapatra, S., & Kayastha, S. (2024). Unveiling the protective role of chitosan in plant defense: A comprehensive review with emphasis on abiotic stress management. Crop Design, 3(4), 100076. https://doi.org/10.1016/j.cropd.2024.100076
Roudbar, M. A., Mohammadabadi, M. R., & Salmani, V. (2015). Epigenetics: A new challenge in animal breeding. Genetics in the Third Millennium, 12(4), 3736-3751. https://www.researchgate.net/publication/281109256_Epigenetics_A_new_Challenge_In_Animal_Breeding
Safaei, S. M. H., Mohammadabadi, M., Moradi, B., Kalashnyk, O., Klopenko, N., Babenko, O., Borshch, O. O., & Afanasenko, V. (2025). Corrigendum: Role of fennel (Foeniculum vulgare) seed powder in increasing testosterone and IGF1 gene expression in the testis of lamb. Gene Expression, 24(4), Article e00020C. https://doi.org/10.14218/GE.2023.00020C
Salarpour, F., & Farahbakhsh, H. (2016). Effects of salicylic acid on some physiological traits, yield and yield components of fennel (Foeniculum vulgare) under drought stress. Iranian Journal of Medicinal and Aromatic Plants Research, 32(2), 216-230. https://doi.org/10.22092/ijmapr.2016.106557
Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology, 299, 152-178. https://doi.org/10.1016/S0076-6879(99)99017-1
Touhidi-Nejad, Z., Farahbakhsh, H., & Maghsoudi, A. (2016). Evaluation of salicylic acid effects on some physiological traits of fenugreek under drought stress. Journal of Plant Process and Function, 5(16), 85-96. https://dor.isc.ac/dor/20.1001.1.23222727.1395.5.16.2.7
Vahabzadeh, M., Chamani, M. M., Dayani, O., Sadeghi, A. A., & Mohammadabadi, M. R. (2021). Effects of sweet marjoram (Origanum majorana) powder on growth performance, nutrient digestibility, rumen fermentation, meat quality and humoral immune response in fattening lambs. Iranian Journal of Applied Animal Science, 11(3), 567-576. https://sanad.iau.ir/Journal/ijas/Article/1023983
Vahabzadeh, M., Chamani, M., Dayani, O., & Sadeghi, A. A. (2020). Effect of Origanum majorana leaf (sweet marjoram) feeding on lamb’s growth, carcass characteristics, and blood biochemical parameters. Small Ruminant Research, 192, Article 106233. https://doi.org/10.1016/j.smallrumres.2020.106233
Yang, H., Mu, J., Chen, L., Feng, J., Hu, J., Li, L., Zhou, J. M., & Zuo, J. (2015). S-nitrosylation positively regulates ascorbate peroxidase activity during plant stress responses. Plant Physiology, 167(4), 1604-1615. https://doi.org/10.1104/pp.114.255216
Yang, Y., Han, X., Liang, Y., Ghosh, A., Chen, J., & Tang, M. (2015). The combined effects of arbuscular mycorrhizal fungi (AMF) and lead (Pb) stress on Pb accumulation, plant growth parameters, photosynthesis, and antioxidant enzymes in Robinia pseudoacacia L. PLoS ONE, 10(12), e0145726. https://doi.org/10.1371/journal.pone.0145726
Yin, H., Kjaer, A., Fretté, X. C., Du, Y., Christensen, L. P., Jensen, M., & Grevsen, K. (2012). Chitosan oligosaccharide and salicylic acid up-regulate gene expression differently in relation to the biosynthesis of artemisinin in Artemisia annua L. Process Biochemistry, 47(11), 1559-1562. https://doi.org/10.1016/j.procbio.2012.05.006
مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 3
شهریور 1405
صفحه 1-26

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