بررسی الگوی بیان ژن‌های ساختاری در مسیر بیوسنتز مونوترپن‌ها و بررسی کیفیت اسانس گیاه دارویی زنیان تحت تأثیر هورمون متیل‌جاسمونات

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم زراعی و اصلاح نباتات، پردیس ابوریحان-دانشگاه تهران، پاکدشت، تهران، ایران

2 گروه علوم زراعی و اصلاح نباتات، پردیس ابوریحان، دانشگاه تهران، پاکدشت، ایران

3 پاکدشت دانشگاه تهران- پردیس ابوریحان- گروه علوم زراعی و اصلاح نباتات

چکیده

هدف: زنیان (.Trachyspermum ammi L) گیاهی است علفی، یکساله از خانواده چتریان (Apiaceae) که در طب سنتی و علوم پزشکی کاربردهای فراوانی دارد. این تحقیق با هدف بررسی الگوی بیان ژن­های ساختاری در مسیر بیوسنتزی مونوترپن­ها و بررسی کیفیت اسانس گیاه دارویی زنیان تحت تأثیر هورمون متیل­جاسمونات انجام شد. ترکیبات شیمیایی اسانس و میزان بیان ژن­های MECPS، DXR و GDs به عنوان ژن­های کلیدی دخیل در مسیر بیوسنتز تیمول پس از اعمال تیمار هورمونی متیل­جاسمونات در 4 زمان­ مورد بررسی قرار گرفت.
مواد و روش: پس از گلدهی، هورمون متیل­جاسمونات 1/0 میلی­مولار به صورت محلول­پاشی روی گیاهان اسپری شد. نمونه­گیری در 4 زمان (8، 24، 48 ساعت پس از هورمون­پاشی و نمونه شاهد اسپری شده با آب مقطر) صورت گرفت. استخراج RNA از بافت گل­آذین گیاه زنیان انجام شد و پس از سنتز cDNA و انجام Real-Time PCR داده­های حاصل از بررسی بیان ژن­های مذکور مورد تجزیه و تحلیل قرار گرفت. جداسازی و شناسایی ترکیبات اسانس نیز با استفاده از دستگاه­های کروماتوگرافی گازی (GC) و کروماتوگرافی گازی متصل به طیف‌سنج جرمی (GC/MS) صورت گرفت. درمجموع، 10 ترکیب در 4 نمونه شناسایی گردید.
نتایج: نتایج حاصل از بررسی ترکیبات اسانس گیاه زنیان نشان داد که با توجه به تغییر مقدار ترکیبات ثانویه موجود در اسانس، میزان تولید آلفاترپینن که پیش­ساز مونوترپن­ها (نظیر تیمول) است، پس از گذشت 24 ساعت از اعمال هورمون متیل­جاسمونات افزایش یافت. همچنین نتایج حاصل از داده­های Real-Time PCR، افزایش بیان هر 3 ژن مذکور پس از 24 ساعت از اعمال هورمون را نشان می­دهد. افزایش بیان ژن­های ذکر شده در مسیر بیوسنتز مونوترپن­ها می­تواند عاملی برای بالا بودن میزان آلفاترپینن در اسانس گل­ها بعد از گذشت 24 ساعت پس از اعمال هورمون باشد.
نتیجه گیری: هورمون متیل­جاسمونات با تحریک مسیر بیوسنتز ترکیب­های ثانویه و همچنین تأثیر بر میزان بیان ژن­های مسیر MEP باعث افزایش تولید مونوترپن­ها یک روز پس از اعمال هورمون می­گردد.

کلیدواژه‌ها


عنوان مقاله [English]

Study the Quantitative Expression Pattern of Some Involved Genes on Monoterpenoid Biosynthesis Pathway and Identification of Essential Compounds Affected by Methyl jasmonate in Ajowan Plant

نویسندگان [English]

  • Seyed Ahmad Sadat Noori 1
  • Mahsa Jamshidi 2
  • Mohammad mahdi Mortazavian 3
1 Department of Agronomy and Plant Breeding Sciences, University of Tehran, Pakdasht, Tehran, Iran
2 Department of agronomy and plant breeding sciences, Tehran university, Pakdasht, Iran
3 Associate Professor, Department of Agronomy and Plant Breeding Sciences, University of Tehran, Pakdasht, Tehran, Iran
چکیده [English]

Objective
Ajowan (Trachyspermum ammi L.) is an annual plant in Apiaceae family which is used in traditional medicine and medical science. This study was carried out to determine the quantitative expression pattern of some genes on monoterpenoid biosynthesis pathway and essential quality in Ajowan treated by methyl jasmonate. Ajowan essential oil compounds and the amount of MECPS, DXR,  and GDs genes expression as main genes related to Thymol biosynthesis pathway in response to methyl jasmonate in 4 different times were analyzed.
 
Materials and methods
After flowering, the 0.1 mM methyl jasmonate hormone was sprayed onto the plants. Samples were taken in 4 times (after 8, 24, and 48 treatment and control treatment). RNA extraction, cDNA synthesis, and Real-Time PCR have done from flowers and the date from these gene expressions were analyzed. Essential chemical compounds were studied by GC-GC/MS. Generally, in 4 samples (samples were taken after 8, 24, and 48 treatment and control treatment), 10 different compounds were distinguished.
 
Results
Based on results, the study of the essential oils of Ajowan showed that the changes of the amount of secondary compounds in the essential oil, the amount of γ-Terpinene (precursor of monoterpenoids like Thymol) went up after 24 hours methyl jasmonate treatment. Due to Real-Time PCR dates, the quantity of all 3 studied genes increased after 24 hours methyl jasmonate treatment. The increasing of these genes expression can be causes the incline level of γ-Terpinene in essential oil.
 
Conclusions
Increasing the amount of 3 mentioned genes expression and also a high amount of γ-Terpinene in essential oils after 24 hours in flowers show that this hormone can affect the gene expression in MEP pathway which causes monoterpenoids production increase after one day of using the hormone.

کلیدواژه‌ها [English]

  • Ajowan (Trachyspermum ammi L.)
  • Essential oil
  • Gene expression
  • methyl jasmonate
  • Secondary metabolites
Aharoni A, Jongsma M, Kim T et al. (2006) Metabolic engineering of terpenoid biosynthesis in plants. Phytochem Rev 5, 49–58.
Ashraf M, Orooj A (2006) Salt stress effects on growth, ion accumulation and seed oil concentration in an arid zone traditional medicinal plant ajwain (Trachyspermum ammi L. Sprague). J Arid Environ 64, 209–220.
Bairwa M, Rajput M, Sachdeva S (2012) Modified Kuppuswamy’s Socioeconomic Scale. Social Researcher Should Include Updated Income Criteria. Indian J Community Med 38, 185–186.
Bettaieb I, Zakhama N, Aidi-Wannes N, Kchouk M, Marzouk B (2009) Water deficit effects on Salvia officinalis fatty acids and essential oils composition. Sci Hortic 120, 271–275.
Bick J, Lange B (2003) Metabolic cross talk between cytosolic and plastidial pathways of isoprenoid biosynthesis: unidirectional transport of intermediates across the chloroplast envelope membrane. Arch Biochem Biophys 415, 146–154.
Brouki milan E, Hassni L, Abdollahi Mandoulakani B et al. (2016)The effect of different concentrations of methyl jasmonate on the activity of antioxidant enzymes and total protein in basil. J Crop Improv 18, 103-115.
Burlat V, Ou32din A, Courtois M, Rideau M, St-Pierre B (2004) Co‐expression of three MEP pathway genes and geraniol 10‐hydroxylase in internal phloem parenchyma of Catharanthus roseus implicates multicellular translocation of intermediates during the biosynthesis of monoterpene indole alkaloids and isoprenoid‐derive. Plant J 38, 131–141.
Coroll C (2011) biosynthesis of the phenolic monoterpenes, thymol and carvacrol, by terpene synthases and cytochrome P450 in oregano and thyme. PhD Thesis at the Max Planck Institute for Chemical Ecology, Jena and the Friedrich-Schiller-University, Jena. 96-108.
Crocoll C, Asbach J, Novak J, Gershenzon J, Degenhardt J (2010) Terpene synthases of oregano (Origanum vulgare L.) and their roles in the pathway and regulation of terpene biosynthesis. Plant Mol Biol 73, 587–603.
Eblagh N, Fateh E, Farzane M, Osfuri M (2014) Effect of Cattle Manure Application, Phosphate Solubilizing Bacteria and Different Phosphrous Levels on Yield and Essence Components of Trachyspermum ammi L. J Agri Sci Sustain Prod, 35-45.
Elyasi R, Majdi M, Bahramnejad b, Mirzaghaderi GH (2015)Expression analysis of genes involved in terpenes biosynthesis in black cumin (Nigella sativa) plants treated with methyl jasmonate. Iranian J Plant Breed Genet Res 24. 54-65.
Ferreira F, David L, Luthria D, Sasaki T, Heyerick A (2010) Flavonoids from Artemisia annua L. as Antioxidants and their potential synergism with artemisinin against malaria and cancer. Molecules 15, 3135–3170.
Ghanadnia M, Hadad R, Zarrinkob F, Sharifi M (2011) Different expression of limonene synthase gene in the organs and developmental stages of Cuminum cyminum L. Iranian J Med Aromatic Plants 27, 495–508.
Ghanati F, Bakhtiyarian S, Abdolmaleki P (2010) Effects of methyl jasmonate on the secondary metabolites of Calendula officinalis L. JMBS 1, 20-30.
Jamshidi E, Ghalavand A, Sefidkon F, Goltaph E (2012) Effects of different nutrition systems (organic and chemical) on quantitative and qualitative characteristics of Fennel (Foeniculum Vulgare Mill.) under water deficit stress. Iranian J Med Aromatic Plants 28, 309-323.
Khajeh M, Yamini Y, Sefidkon F, Bahramifar N (2004) Comparison of essential oil composition of Carum copticum obtained by supercritical carbon dioxide extraction and hydrodistillation methods. Food Chem 86, 587–591.
Krishnamoorthy V, Madalageri M (1999) Bishop weed (Trachyspermum ammi L.): an essential crop for north Karnataka. Med Aromat Plant Sci Biotech 21, 996–998.
Lawrence B (2000) Ajowan oil compositional and bibligaphilical data. Perfume Flavourist 19, 13–20.
Mahmoud S, Croteau R (2002) Strategies for transgenic manipulation of monoterpene biosynthesis in plants. Trends Plant Sci 7, 366–373.
Majidi M, Karimzadeh G, Malboobi M (2014) Spatial and developmental expression of key genes of terpene biosynthesis in Tanacetum parthenium. Biol Plantarum 58, 379–384.
Majnoon Hosseini N, Davazdah-Emami S (2007) Crops and the Production of Medicinal Plants and Spices. (K. Peter, Ed.) (Malhotra,). Tehran University Publications.
Mirzahosseini SM, Sadat-Noori SA, Amanzadeh Y, Javid MG, Howyzeh MS (2017) Phytochemical assessment of some native ajowan (Therachyspermum ammi L.) ecotypes in Iran. Ind Crops Prod 105, 142-147.
Phillipson JD (2007) Phytochemistry and pharmacognosy. Phytochem 68, 2960–2972.
Ramak P, Kazempour Osaloo S, Ebrahimzadeh H, Sharifi M, Behmanesh M (2012) Expression of gene 1-deoxy-D-xylulose 5-phosphate reductoisomerase and its relation to monoterpene Carvacrol biosynthesis in Satureja khuzestanica. J Plant Res (Iran J Biol) 27, 622-634.
Ramanujam S, Joshi B, Saxena MB (1964) Extent and randomness of cross-pollination in some umbelliferous spices of India. Indian J Genet Plant Breed 24, 62–67.
Rebey B, Jabri-Karoui I, Hamroumi-Sellami I, Bourgou S, Limam F, & Marzouk B (2012) Effect of drought on the biochemical composition and antioxidant activities of cumin (Cuminum cyminum L.) seeds. Ind Crops Prod 36, 238–245.
Rezaei-Chiyaneh E (2011) Effects of different irrigation treatments on essential oil accumulation, its composition and some ecophysiological traits in Fennel (Foeniculum vulgare). PhD thesis Department of Plant of Eco-physiological, Faculty of Agriculture, university of Tabriz, Tabriz. 329-338.
Sadat-Noori S, Norouzi M, Karimzadeh G, Shirkool K, Niazian, M (2017) Effect of colchicine-induced polyploidy on morphological characteristics and essential oil composition of ajowan (Trachyspermum ammi L.). Plant Cell Tissue Organ Cult (PCTOC), 1–9.
Saydpour F, Sayyari M (2016) Impact of methyl jasmonate on Enhancing Chilling Tolerance of Cucumber (Cucumis sativus L.) Seedlings. J Crop Prod Proc (JCPP) 6, 47-59.
Soltani Howyzeh M, Sadat-Noori SA, Shariati J (2018) Essential oil profiling of Ajowan (Trachyspermum ammi) industrial medicinal plant. Ind Crops Prod 119, 255-259.
Soltani Howyzeh M, Sadat-Noori SA, Shariati J, Amiripour M (2018) Comparative transcriptome analysis to identify putative genes involved in thymol biosynthesis pathway in medicinal plant Trachyspermum ammi L. Sci Rep 8. 1-19.
Soltani Howyzeh M, Sadat-Noori SA, Shariati J, Amiripour M (2018) Identification of terpenoid backbone biosynthetic pathway genes in Ajowan (Trachyspermum ammi L.) by RNA-Seq. Modern Genet J (MGJ) 3, 133-141.  
Trapp S, Croteau R (2001) Genomic organization of plant terpene synthases and molecular evolutionary implications. Genet 158, 811–832.
Veau B, Courtois M, Oudin A, Chenieux J, Rideau M, Clastre M (2000) Cloning and expression of cDNAs encoding two enzymes of the MEP pathway in Catharanthus roseus Biochim Biochim Biophys. Biochim Biophys, 159–163.