Expression analysis of the key genes involved in the biosynthesis pathway of Carvacrol in Satureja mutica Fisch and CA mey under drought stress

Document Type : Research Paper

Authors

1 MSc Student, Department of Plant Production and Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

2 Assistant Professor, Department of Plant Production and Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

3 Key Laboratory of Plant Resources/Beijing Botanical Garden, Institute of Botany, Chinese Academy of Sciences, Beijing, China

Abstract

Objective
Medicinal plants have long been recognized as a rich source of essential oils and have been cultivated for a variety of uses since ancient times. White savory (Satureja mutica Fisch & CA mey) is one of the medicinal plants belonging to the mint family with high content of carvacrol in its essential oil. A large body of literature has suggested that abiotic stresses, especially drought stress, could considerably boost the production of essential oils in plants. This study aimed to evaluate the effect of drought stress on the expression of genes affecting the biosynthesis of the main component of white savory essential oil carvacrol.
Materials and methods
For this goal, 20 pot-planted seedlings of white savory at the 6-8 leaf stage were subjected in a greenhouse to two conditions of control and drought stress (10 pots for each condition) with irrigation regimes of 100% and 30% of field capacity, respectively. To ensure the implementation of stress on plants and to determine the appropriate time for sampling, leaf proline and relative water content of plants were measured.
Results
The results of both proline and relative water content tests revealed a significant statistical difference between the control and stress conditions in white savory at 30% of field capacity. The KEGG (Kyoto Encyclopedia of Genes and Genomes) database was used to obtain information on the carvacrol biosynthesis pathway. The three key genes geranyl diphosphate synthase (gpps), terpene synthase (tps), and limonene hydroxylase (lh) were chosen for gene expression analysis. Due to the lack of genomic information on white savory, the sequence information of the genes in the closest genera within the family mint was used to design primers. The result of melting curve analysis from the real-time quantitative PCR (qPCR) assay confirmed the accuracy of the primer design. The raw qPCR data normalized to the housekeeping gene Actin, then differential expression analysis was performed using the delta-delta Ct method. qPCR result showed that expression of the genes gpps, tps, and lh were increased in stressed plants proximately 20, 18, and 50 times, compared to those in control, respectively.
Conclusions
The conclusion that must be drawn from the data available is that water stress leads to the process of plant response to stress and changes the expression of genes involved in the biosynthesis pathway of the active substance carvacrol. 

Keywords

References

الیاسی ریزان، مجدی محمد، بهرام‌نژاد بهمن، میرزا قادری قادر. (1394). بررسی الگوی بیان ژن‌های دخیل در مسیر بیوسنتزی مونوترپن‌‌ها و تری‌ترپن‌ها تحت تأثیرتیمار اسیدسالیسیلیک در گیاه دارویی سیاه‌دانه. تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران، 23(2)، 174-164.
حبیبی شکوفه، قادری اردشیر، فاتحی فواد (1396). بررسی بیان نسبی ژنهای کلیدی مسیر بیوسنتزی تیمول تحت تنش سرمایی در گیاه آویشن باغی رقم ’واریکو3’) (Thymus vulgaris cv. Varico 3’) با استفاده از PCR در زمان واقعی. فصلنامه گیاهان دارویی، 4(11)، 50-58.
حسینی حسیبه، موسوی فرد صادق، فاتحی فواد، قادری اردشیر (1395) تغییرات فیتوشیمیایی و صفات مرفو- فیزیولوژیکی گیاه دارویی آویشن باغی (Thymus vulgaris cv. Varico 3’) تحت تنش شوری. فصلنامه گیاهان داروی، 16(61)، 33-22.
رهایی مهدی، نقوی محمدرضا، علیزاده هوشنگ، و همکاران (1389) بررسی الگوی بیان ژن‌های MYB در گندم (Triticum aestivum L.) تحت دو تنش کوتاه‌مدت شوری و سرما با استفاده از راهکار RT-PCR کمی. علوم گیاهان زراعی ایران، 41(3)، 433-346.
صالحی ارجمند حسین (1384) تأثیر تنش‌های محیطی در افزایش متابولیت‌های ثانویه در گیاهان. همایش ملی توسعه پایدار گیاهان دارویی. انتشارات مؤسسه تحقیقات جنگل‌ها و مراتع، کرج، ص. 307-305.
فرزانه اکرم، غنی عسکر، عزیزی مجید (1391)  تأثیر تنش آبی بر ویژگی‌های ظاهری، عملکرد و درصد اسانس در گیاه ریحان (رقم کشکنی لو لو) (keshkeni luvelou). پژوهش‌های تولید گیاهی، 17(1)، 111-103.
محمدی ولی‌اله، مندک بهنام، هادیان جواد، زینالی خانقاه حسن (1399). اثر دما بر متابولیت‌ها و بیان برخی از ژن‌های مسیر بیوسنتز تیمول و کارواکرول در آویشن باغی (Thymus vulgaris). علوم گیاهان زراعی ایران، 51(1)، 205-195.
محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده از‌Real Time PCR . مجله بیوتکنولوژی کشاورزی،  12(1)، 177-192.
محمدآبادی محمدرضا، سفلایی، محمد (1399) پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی، 12(3)، 208-191.
محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافتهای مختلف گوسفند کرمانی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی، 10(3)، 122-111.
References
Abdalla MM, El-Khoshiban NH (2007) The influence of water stress on growth, relative water content, photosynthetic pigments, some metabolic and hormonal contents of two Triticum aestivum cultivars. J Appl Sci Res 3(12), 2062-2074.
Ashrafi M, Azimi-Moqadam MR, MohseniFard E et al. (2022). Physiological and molecular aspects of two thymus species differently sensitive to drought stress. BioTech 11(8), 1-11.
Baher ZF, Mirza M, Ghorbanli M, Bagher Rezaii M (2002). The influence of water stress on plant height, herbal and essential oil yield and composition in Satureja hortensis L. Flavour Fragr J 17(4), 275-277.‏
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of proline for water stress studies. Plant Soil 39, 205-208.
Bose SK, Yadav RK, Mishra S et al. (2013) Effect of gibberellic acid and calliterpenone on plant growth attributes, trichomes, essential oil biosynthesis and pathway gene expression in differential manner in Mentha arvensis L. Plant Physiol Biochem 66, 150-158.
Bouvier F, Suire C, d'Harlingue A et al. (2000) Molecular cloning of geranyl diphosphate synthase and compartmentation of monoterpene synthesis in plant cells. Plant J 24(2), 241-252.
Carretero-Paulet L, Ahumada I, Cunillera N et al. (2002) Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiol 129(4), 1581-1591.
Chai Q, Gan Y, Zhao C et al. (2016) Regulated deficit irrigation for crop production under drought stress: A review. Agron Sustain Dev 36, 1-21.
Dubey VS, Bhalla R, Luthra R (2003) An overview of the non-mevalonate pathway for terpenoid biosynthesis in plants. J Biosci 28, 637-646.
Elyasi R, Majdi M, Bahramnejad B, Mirzaghaderi GH (2015) Expression pattern analysis of genes involved in the biosynthetic pathway of monoterpenes and triterpenes in black cumin (Nigella sativa) plants treated with salicylic acid. Iran J Rangeland Forest Plant Breed Genet Res 23(2), 164-174. )In Persian(
Farooq M, Hussain M, Wahid A, Siddique KHM (2012).Drought Stress in Plants: An Overview. In: Aroca, R. (eds) Plant Responses to Drought Stress. Springer, Berlin, Heidelberg.
Farzaneh A, Ghani A, Azizi M (2010) The effect of water stress on morphological characteristic and essential oil content of improved sweet basil (Ocimum basilicum L). J Plant Production 17(1), 103-111. )In Persian(
Habibi Sh, Qaderi A, Fatehi F (2017) The study of relative expression of key genes of thymol biosynthesis pathway in thymus vulgaris cv.‘Varico 3’under cold stress using real-time PCR. J Med Plants 4(11), 50-58. )In Persian(
Hadian J, Akramian M, Heydari H et al. (2012) Composition and in vitro antibacterial activity of essential oils from four Satureja species growing in Iran. Nat Prod Res 26(2), 98-108.
Haileselasie TH, Teferii G (2012) The effect of salinity stress on germination of chickpea (Cicer arietinum L.) land race of Tigray. Curr Res J Biol Sci 4, 578-583.‏
Hassan FA, Ali EF (2014) Impact of different water regimes based on class-A pan on growth, yield and oil content of Coriandrum sativum L. plant. Journal of the Saudi Society of Agricultural Sciences 13, 155-161.
Hassanpour S, Heppler GR. (2014). Step-by-step simplification of the micropolar elasticity theory to the couple-stress and classical elasticity theories. Proceedings of the ASME 2014 International Mechanical Engineering Congress and Exposition IMECE2014. November 14-20, 2014, Montreal, Quebec, Canada.‏
Hosseini H, Mousavi-Fard S, Fatehi F, Qaderi A. (2017) Changes in phytochemical and morpho-physilogical traits of thyme (Thymus vulgaris CV Varico 3) under different salinity levels. J Med Plants 16(61), 22-33. (In Persian)
Jamzad Z (2009) Thymus and Satureja species of Iran. Research Institute of Forests and Rangelands, Tehran, Iran. (In Persian)
Karimi E, Ghasemnezhad A, Hadian J, Ghorbanpour M (2016) Assessment of essential oil constituents and main agro-morphological variability in Satureja mutica populations. Braz J Bot 39, 77-85.
Karunanithi PS, Berrios DI, Wang S et al. (2020) The foxtail millet (Setaria italica) terpene synthase gene family. Plant J 103(2), 781-800.
Khorasaninejad S, Mousavi A, Soltanloo H et al. (2011) The effect of drought stress on growth parameters, essential oil yield and constituent of Peppermint (Mentha piperita L.). J Med Plants Res 5(22), 5360-5365.
Masoudzadeh SH, Mohammadabadi M, Khezri A et al. (2020) Effects of diets with different levels of fennel (Foeniculum vulgare) seed powder on DLK1 gene expression in brain, adipose tissue, femur muscle and rumen of Kermani lambs. Small Rumin Res 193, e106276.
Miranshahi B, Sayyari M (2016) Methyl jasmonate mitigates drought stress injuries and affects essential oil of summer savory. J Agr Sci Tech 18(6), 1635-1645.
Mohammadabadi M, Masoudzadeh SH, Khezri A et al. (2021) Fennel (Foeniculum vulgare) seed powder increases Delta-Like Non-Canonical Notch Ligand 1 gene expression in testis, liver, and humeral muscle tissues of growing lambs. Heliyon 7‌(12), e08542.
Mohammadabadi M, Soflaei M (2020) Tissue-specific mRNA expression profile of BMP15 gene in goat. Agricultural Biotechnology Journal 12(3), 191-208. (In Persian)
Mohammadabadi MR (2020) Expression of ESR1 gene in Raini Cashmere goat using Real Time PCR. Agricultural Biotechnology Journal 12‌(1), 177-192. (In Persian)
Mohammadabadi MR, Kord M, Nazari M (2018) Studying expression of leptin gene in different tissues of Kermani Sheep using Real Time PCR. Agric Biotechnol J 10(3), 111-122 (In Persian)
Mohammadabadi MR, Tohidinejad F (2017) Charachteristics determination of Rheb gene and protein in Raini Cashmere goat. Iranian Journal of Applied Animal Science 7(2), 289-295.
Mohammadi V, Mondak B, Hadian J, Zeinali Khanghah H (2020) The impact of temperature on metabolites and the expression of genes involved in thymol and carvacrol biosynthesis pathway in Thymus vulgaris. Iran J Field Crop Sci 51(1), 195-205. (In Persian)
Nemeth-Zambori E, Szabo K, Pluhar Z et al. (2016) Changes in biomass and essential oil profile of four Lamiaceae species due to different soil water levels. Journal of Essential Oil Research 28(5), 1-9.
Peltzer M, Wagner JR, Jimenez A (2007) Thermal characterization of uhmwpe stabilized with natural antioxidants. J Therm Anal Calorim 87, 493-497.
Pirzad A, Shakiba MR, Zehtab-Salmasi S, et al. (2011) Effect of water stress on leaf relative water content, chlorophyll, proline and soluble carbohydrates in Matricaria chamomilla L. J Med Plant Res 5(12), 2483-2488.‏
Poulose AJ, Croteau R (1978a) Biosynthesis of aromatic monoterpenes: Conversion of γ-terpinene to p-cymene and thymol in Thymus vulgaris L. Arch Biochem Biophys 187(2), 307-314.
Poulose AJ, Croteau R (1978b) γ-Terpinene synthetase: A key enzyme in the biosynthesis of aromatic monoterpenes. Arch Biochem Biophys 191, 400-411.
Rahaei M, Naghavi M, Alizadeh H et al. (2010) The gene expression analysis of MYB transcription factors in wheat (Triticum aestivum L.) during short term salinity and cold stresses through quantitative RT-PCR. Iran J Field Crop Sci 41(3), 433-346. (In Persian)
Rahimi Y, Taleei A, Ranjbar M (2017a) Changes in the expression of key genes involved in the biosynthesis of menthol and menthofuran in Mentha piperita L. under drought stress. Acta Physiol Plant 39, 1-9.
Rahimi Y, Taleei A, Ranjbar M (2017b) Investigation of lim, medh gene expression level and morpho-physiological changes of Mentha piperiat L. in response to drought. Iran J Field Crop Sci 48(2), 507-516.
Rai V, Vajpayee P, Singh SN, Mehrotra S (2004) Effect of chromium accumulation on photosynthetic pigments, oxidative stress defense system, nitrate reduction, proline level and eugenol content of Ocimum tenuiflorum L. Plant Science 167(5), 1159-1169.
Saei‐Dehkordi S, Fallah AA, Heidari‐Nasirabadi M, Moradi M (2012) Chemical composition, antioxidative capacity and interactive antimicrobial potency of Satureja khuzestanica Jamzad essential oil and antimicrobial agents against selected food‐related microorganisms. Int J Food Sci Technol 47(8), 1579-1585.
Saharkhiz MJ, Zomorodian K, Taban A et al. (2016) Chemical composition and antimicrobial activities of three Satureja Species against food-borne pathogens. J Essential Oil Bearing Plants 19, 1984-1992.
Salehi Arjomand H (2005) The effect of environmental stresses on the increase of secondary metabolites in plants. National Conference on Sustainable Development of Medicinal Plants. Publications of Forests and Rangelands Research Institute, Karaj, p. 307-305. (In Persian)
Sanchez SR (1998) Turgor maintenance, osmotic adjustment and soluble sugar and proline accumulation in 49 pea cultivars in response to water stress. Field Crops Res 59, 225-235.
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2021) Correlation between insulin-like growth factor 1 gene expression and fennel (Foeniculum vulgare) seed powder consumption in muscle of sheep. Anim Biotechnol 33, 1-11.
Usano-Alemany J, Pala-Paul J, Herraiz-Penalver D (2014) Temperature stress causes different profiles of volatile compounds in two chemotypes of Salvia lavandulifolia Vahl. Biochem Syst Ecol 54, 166-171.‏
Yang WC, Bao HY, Liu YY et al. (2018) Depsidone derivatives and a cyclopeptide produced by marine fungus Aspergillus unguis under chemical induction and by its plasma induced mutant. Molecules 23(9), 2245.
Zarshenas MM, Krenn L (2015) A critical overview on Thymus daenensis Celak. phytochemical and pharmacological investigations. J Integr Med 13(2), 91-98.
Zhou HC, Shamala LF, Yi XK et al. (2020) Analysis of terpene synthase family genes in Camellia sinensis with an emphasis on abiotic stress conditions. Sci Rep 10, 933.
Agricultural Biotechnology Journal
Volume 14, Issue 3
September 2022
Pages 151-170

Files

Share

How to cite

Statistics