Comparison of expression pattern of some artemisinin biosynthesis related genes and phytochemical profile in Artemisia fragrans and Artemisia annua species

Document Type : Research Paper

Authors

1 Ph.D. Student, Department of Plant Breeding and Biotechnology, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

2 Assistant Professor, Department of Plant Breeding and Biotechnology, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

3 Associate Professor, Department of Plant Breeding and Biotechnology, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

4 Assistant Professor, Seed and Plant Improvement Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

5 Department of Plant Breeding and Biotechnology, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

Abstract

Objective
So far around 500 species of Artemisia have been found in different regions of the world, and among them 34 species are endemic to Iran. It has been evidenced that Artemisinin is one of the medicinal compounds found in Artemisia that has various medical properties, especially antimalarial. The main objective of the present study was to comprise phytochemical profile and expression pattern of some genes related to artemisinin biosynthesis pathway in A. fragrans and A. annua species.
Materials and methods
In the present study, the accumulated artemisinin content in A. fragrans and A. annua species were detected using the high-performance liquid chromatography (HPLC) technique. Moreover, phytochemical compounds in leave tissue were identified using the gas chromatography–mass spectrometry (GC-MS) technique. The gene expression patterns for some artemisinin biosynthesis related genes including 4FPSF, DBR2, HMGR1, HMGR2, WIRKY, ADS, DXS, and SQS were comprised in two studied species. In each species, association between artemisinin content and the relative expression of investigated genes were determined through correlation analysis. All statistical analysis was performed based on three replication using R software.
Results
Based on obtained results, there was observed a significant difference between two Artemisia species in terms of artemisinin content, and the highest content was recorded for A. fragrans. Using GC-MS analysis, in general 26 and 20 phytochemical compounds were identified in A. fragrans and A. annua species, respectively. Among identified compounds, eight compounds were similar in both species. Moreover, some compounds such as Comphor, 1,8-Cineole, 4-Terpineol, and Pinocarvone were most important. According to the gene expression analysis, the highest relative expression of 4FPSF, ADS, and DXS genes were recorded in A. annua, while the highest numbers of transcripts for SQS, HMGR1, HMGR2, DRB2, and WIRKY genes were estimated in A. fragrans. The results of correlation analysis for both species showed that artemisinin content significantly and positively correlated with the expression of ADS, DBR2, DXS, and HMGR1 genes. However, in A. annua species, the SQS gene was not expressed and there was no correlation between these genes and artemisinin content.
Conclusions
In general, the obtained results revealed a significant difference between two studied Artemisia species in terms of the artemisinin content and other phytochemical compounds. Furthermore, our results indicated that A. fragrans could be used as an ideal source for extract artemisinin and other compounds. Hence, conducting other supplementary studies on this species is recommended.

Keywords


آروین پویا، فیروزه رعنا (1401) بررسی کیفیت، کمیت و فعالیت آنتی اکسیدانی مواد مؤثره اسانس دو گونه دارویی درمنه کپت داغی (Artemisia kopetdaaghensis Krasch.) و درمنه سیبریایی (A. siberi Besser.). فرایند و کارکرد گیاهی، 50، 211-199.
پاشازانوسی محمدباقر (1401) جداسازی و شناسایی ترکیبات شیمیایی روغن اسانسی دو گونه درمنه (آرتمیزیا) در شمال ایران با استفاده از کروماتوگرافی گازی-طیف‌سنج جرمی. شیمی کوانتومی و اسپکتروسکوپی. 40، 30-17.
جعفری احمدآبادی سید علی اصغر، عسکری­همت حشمت­اله، محمدآبادی محمدرضا (1402) تاثیر شاهدانه بر بیان ژن DLK1 در بافت‌ قلب بره‌های کرمانی. مجله بیوتکنولوژی کشاورزی، 15(1)، 217-234.
شکری سمیرا، خضری امین، محمدآبادی محمدرضا، خیرالدین حمید (1402) بررسی بیان ژن MYH7  در بافت‌های ران، دست و راسته بره‌های پرواری نژاد کرمانی. مجله بیوتکنولوژی کشاورزی، 15(2)، 217-236.شرفی ع، هاشمی­سهی ه، کاظمی­تبار ک (1385) مروری بر کشت بافت، باززایی و انتقال ژن به Artemisia annua L.. فصلنامه گیاهان دارویی، 20، 10-1.
References
Alam P, Kamaluddin Khan MA, Mohammad A, et al. (2014) Enhanced artemisinin accumulation and metabolic profiling of transgenic Artemisia annua L. plants over-expressing by rate-limiting enzymes from isoprenoid pathway. J Plant Interact 9, 655–665.
Aquil S, Husaini AM, Abdin MZ, Rather GM (2009) Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants. Planta Med 75, 1453–1458.
Arsenault P, Vail DR, Wobbe KK, Weathers PJ (2010) Effect of sugars on artemisinin production in Artemisia annua L.: Transcription and metabolite measurements. Molecules 15, 2302–2318.
Arvin P, Firouzeh R (2022) Study of quality, quantity and antioxidant activity of essential oils of two medicinal species of Artemisia kopetdaghensis Krasch. and A. sieberi Besser. Plant Process Funct 11, 199–212.
Barazandeh A, Mohammadabadi MR, Ghaderi-Zefrehei M, Nezamabadipour H (2016a) Predicting CpG Islands and Their Relationship with Genomic Feature in Cattle by Hidden Markov Model Algorithm. Iran J Appl Anim Sci 6 (3), 571-579.
Barazandeh A, Mohammadabadi MR, Ghaderi-Zefrehei M, Nezamabadipour H (2016b) Genome-wide analysis of CpG islands in some livestock genomes and their relationship with genomic features. Czech J Anim Sci 61, e487.
Benyagoub E, Nabbou N, Dine A (2020) Antimicrobial effect of Quercus robur L. leaves selective extracts from the Mezi Mountain of Djeniene Bourezg (West of Algeria). Curr Bioact Compd 16, 1181–1190.
Bordbar F, Mohammadabadi M, Jensen J, et al. (2022) Identification of candidate genes regulating carcass depth and hind leg circumference in simmental beef cattle using Illumina Bovine Beadchip and next-generation sequencing. Animals 12(9), e1103.
Czechowski T, Larson TR, Catania TM, et al. (2018) Detailed phytochemical analysis of high-and low artemisinin-producing chemotypes of Artemisia annua. Front Plant Sci 9, e641.
Efferth T, Zacchino S, Georgiev MI, et al. (2015) Nobel Prize for artemisinin brings phytotherapy into the spotlight. Phytomedicine 22, 1–3.
Ghassemi G, Dehghan Nayeri F, Hosseini R (2021) The effects of chitosan nanoparticles on genes expression of artemisinin synthase in suspension culture of Artemisia annua L: A comparative study. Int J Adv Biol Biomed Res 2021, 214–227.
Guo XX, Yang XQ, Yang RY, Zeng QP (2010) Salicylic acid and methyl jasmonate but not Rose Bengal enhance artemisinin production through invoking burst of endogenous singlet oxygen. Plant Sci 178, 390–397.
Hamidi F, Karimzadeh G, Rashidi Monfared G, Salehi M (2018) Assessment of Iranian endemic Artemisia khorassanica: karyological, genome size, and gene expressions involved in artemisinin production, Turk J Biol 42, 322–333.
Han J, Wang H, Lundgren A, Brodelius PE (2014) Effects of overexpression of AaWRKY1 on artemisinin biosynthesis in transgenic Artemisia annua plants. Phytochem 102, 89–96.
Jafari Ahmadabadi SAA, Askari-Hemmat H, Mohammadabadi M, et al. (2023) The effect of Cannabis seed on DLK1 gene expression in heart tissue of Kermani lambs. Agric Biotechnol J 15 (1), 217-234 (In Persian).
Kim SH, Chang YJ, Kim SU (2008) Tissue specificity and developmental pattern of amorpha-4, 11-diene synthase (ADS) proved by promoter-driven GUS expression in the heterologous plant Arabidposis thaliana. Planta Med 74, 188–193.
Liu H, Qiu N, Ding H, Yao R (2008) Polyphenols contents and antioxidant capacity of 68 Chinese herbals suitable for medical or food uses, Food Res Int 41, 363–370.
Liu Y, Ye HC, Wang H, Li GF (2003) Molecular cloning Escherichia coli expression and genomic organization of squalene synthase gene from Artemisia annua. Acta Bot Sin 45, 608–613.
Mannan A, Ahmed I, Arshad W, Asim MF, Qureshi RA, Hussain I, Mirz B (2010) Survey of artemisinin production by diverse Artemisia species in northern Pakistan. Malaria J 9, 310.
Masoudzadeh SH, Mohammadabadi MR, Khezri A, et al. (2020) Dlk1 gene expression in different Tissues of lamb. Iran J Appl Anim Sci 10, 669-677.
Matsushita Y, Kang W, Charlwood BV (1996) Cloning and analysis of a cDNA encoding famesyl diphosphate synthase from Artemisia annua. Gene 172, 207–209.
Mohadjerani M, Rasolizadeh A (2020) Evaluation of kinetic parameters of acetylcholinesterase from the bovine brain and its inhibition with Mentha pulegium L., extract. J Ilam Univ 27, 74–87.
Mohamadipoor L, Mohammadabadi M, Amiri Z, et al. (2021) Signature selection analysis reveals candidate genes associated with production traits in Iranian sheep breeds. BMC Vet Res 17 (1), 1-9.
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.
Mohammadinejad F, Mohammadabadi M, Roudbari Z, Sadkowski T (2022) Identification of Key Genes and Biological Pathways Associated with Skeletal Muscle Maturation and Hypertrophy in Bos taurus, Ovis aries, and Sus scrofa. Animals 12 (24), 3471.
Mozafarian V (1989) Investigation and identification of Artemisia spp. in Iran. MSc thesis, Faculty of Natural Resources, Tehran University.
Nguyen KT, Arsenault RR, Weathers PJ (2011) Trichomes + roots + ROS = artemisinin: regulating artemisinin biosynthesis in Artemisia annua L., In Vitro Cell. Dev. Biol-Plant. 47, 329–338.
Numonov S, Sharopov F, Salimov A, Sukhrobov P, et al. (2019) Assessment of artemisinin contents in selected Artemisia species from Tajikistan (central Asia), Medicines 6, e23.
Olofsson L, Engström A, Lundgren A, Brodelius PE (2011) Relative expression of genes of terpene metabolism in different tissues of Artemisia annua L. BMC Plant Biol 11, 45–57.
Pashazanosi M (2023) Solvent-free microwave extraction of essential oil of two artemisia species (A. spicigera and A. chamaemelifolia) (from from north of Iran. J Quant Chem Spect 40, 30–37.
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR.
Nucl Acids Res 29, e45.
Ranjbar M, Naghavi MR, Alizadeh H, Soltanloo H (2015) Expression of artemisinin biosynthesis genes in eight Artemisia species at three developmental stages. Ind Crop Prod 76, 836–843.
Safaei SMH, Dadpasand M, Mohammadabadi M, et al. (2022) An Origanum majorana Leaf Diet Influences Myogenin Gene Expression, Performance, and Carcass Characteristics in Lambs. Animals 13 (1), e14.
Salehi M, Karimzadeh G, Naghavi MR, et al. (2018b) Expression of artemisinin biosynthesis and trichome formation genes in five Artemisia species. Ind Crop Prod 112, 130–140.
Salehi M, Karimzadeh G, Naghavi MR, et al. (2018a) Expression of key genes affecting artemisinin content in five Artemisia species, Sci Rep 8, 12659.
Salih AM, Qahtan AA, Al-Qurainy F (2023) Phytochemicals identification and bioactive compounds estimation of Artemisia species grown in Saudia Arabia, Metabolites 13, e443.
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2022) Effect of Fennel (Foeniculum Vulgare) Seed Powder Consumption on Insulin-like Growth Factor 1 Gene Expression in the Liver Tissue of Growing Lambs. Gene Express 21 (2), 21-26.
Sharafi A, Hashemi Sohi H, Kazemi Tabar K (2006) Overview on Tissue Culture, Regeneration and Gene Transfornation to Artemisia annua L. J Med Plants 5, 1-10.
Shokri S, Khezri A, Mohammadabadi M, Kheyrodin H (2023). The expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of the Kermani breed. Agric Biotechnol J 15 (2), 217-236 (In Persian).
Verdian-rizi MR, Sadat-Ibrahimi E, Hajiakhoondi A (2008) Chemical composition and antimicrobial activity of Artemisia annua L. essential oil from Iran. J Med Plant 7, 59-62.
Wang H, Liu Y, Chong K, et al. (2007) Earlier flowering induced by over-expression of CO gene does not accompany increase of artemisinin biosynthesis in Artemisia annua. Plant Biol 9, 442–446.
Wang H, Ma C, Li Z, et al. (2009) Effects of exogenous methyl jasmonate on artemisinin biosynthesis and secondary metabolites in Artemisia annua L. Ind Crops Prod 31, 214–218.
Watson LE (2002) Molecular phylogeny of subtribe Artemisiinae (Asteracea), including Artemisia and its allied and segregate genera. BMC Evol Biol 2, e17.
Weathers PJ, Elkholy S, Wobbe KK (2006) Artemisinin: the biosynthetic pathway and its regulation in Artemisia annua, a terpenoid-rich species, In Vitro Cell. Dev-Pl. 42, 309–317.
Zia M, Mannan A, Chaudhary MF (2007) Effect of growth regulators and amino acids on artemisinin production in the callus of Artemisia absinthium. Pak J Bot 39, 799–805.Akyol I, Comlekcioglu U, Kar B, et al. (2009) Cloning of a xylanase gene xyn2A from rumen fungus Neocallimastix sp. GMLF2 in Escherichia coli and its partial characterization. Biologia 64, 664–670.