Main conclusion
Morphological, phytochemical, and transcriptome analyses revealed candidate genes involved in the biosynthesis of volatile monoterpenes and development of glandular trichomes in Monarda citriodora.
Abstract
Monarda citriodora Cerv. ex Lag. is a valuable aromatic plant due to the presence of monoterpenes as major constituents in its essential oil (EO). Thus, it is of sheer importance to gain knowledge about the site of the biosynthesis of these terpenoid compounds in M. citriodora, as well as the genes involved in their biosynthesis. In this study, we studied different types of trichomes and their relative densities in three different developmental stages of leaves, early stage of leaf development (L1), mid-stage of leaf development (L2), and later stage of leaf development (L3) and the histochemistry of trichomes for the presence of lipid and terpenoid compounds. Further, the phytochemical analysis of this plant through GC–MS indicated a higher content of monoterpenes (thymol, thymoquinone, γ-terpinene, p-cymene, and carvacrol) in the L1 stage with a substantial decrease in the L3 stage of leaf development. This considerable decrease in the content of monoterpenes was attributed to the decrease in the trichome density from L1 to L3. Further, we developed a de novo transcriptome assembly by carrying out RNA sequencing of different plant parts of M. citriodora. The transcriptome data revealed several putative unigenes involved in the biosynthesis of specialized terpenoid compounds, as well as regulatory genes involved in glandular trichome development. The data generated in the present study build a strong foundation for further improvement of M. citriodora, in terms of quantity and quality of its essential oil, through genetic engineering.
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Data availability
All datasets generated for this study are included in the manuscript as its supplementary data.
Abbreviations
- EO:
-
Essential oil
- MEP:
-
2C-methyl-D-erythritol-4-phosphate pathway
- MVA:
-
Mevalonic acid pathway
References
Ali MY, Akter Z, Mei Z, Zheng M, Tania M, Khan MA (2021) Thymoquinone in autoimmune diseases: therapeutic potential and molecular mechanisms. Biomed Pharmacother 134:111157. https://doi.org/10.1016/j.biopha.2020.111157
Anders S, Huber W (2010) Differential expression analysis for sequence count data. Genome Biol 11:R106. https://doi.org/10.1186/gb-2010-11-10-r106
Baser KHC, Buchbauer G (2009) Handbook of essential oils: Science, technology, and applications. CRC Press, Baco Raton
Bhatt A, Naidoo Y, Nicholas A (2010) An investigation of the glandular and non-glandular foliar trichomes of Orthosiphon labiatus N.E.Br. [Lamiaceae]. N Z J Bot 48(3–4):153–161. https://doi.org/10.1080/0028825X.2010.500716
Butt MS, Imran M, Imran A, Arshad MS, Saeed F, Gondal TA, Shariati MA, Gilani SA, Tufail T, Ahmad I, Rind NA (2021) Therapeutic perspective of thymoquinone: a mechanistic treatise. Food Sci Nutr 9(3):1792–1809. https://doi.org/10.1002/fsn3.2070
Chalvin C, Drevensek S, Dron M, Bendahmane A, Boualem A (2020) Genetic control of glandular trichome development. Trends Plant Sci 25(5):477–487. https://doi.org/10.1016/j.tplants.2019.12.025
Choi JS, Kim ES (2013) Structural features of glandular and non-glandular trichomes in three species of Mentha. Appl Microsc 43(2):47–53. https://doi.org/10.9729/am.2013.43.2.47
Deepika, Singh A, Chaudhari AK, Das S, Dubey NK (2020) Nanoencapsulated Monarda citriodora Cerv. ex Lag. essential oil as potential antifungal and antiaflatoxigenic agent against deterioration of stored functional foods. J Food Sci Technol 57:2863–2876. https://doi.org/10.1007/s13197-020-04318-4
Delker DA, Geter DR, Roop BC, Ward WO, Ahlborn GJ, Allen JW, Nelson GM, Ouyang M, Welsh W, Chen Y, O’Brien T (2009) Oncogene expression profiles in K6/ODC mouse skin and papillomas following a chronic exposure to monomethylarsonous acid. J Biochem Mol Toxicol 23(6):406–418. https://doi.org/10.1002/jbt.20304
Demarco D (2017) Histochemical analysis of plant secretory structures. Methods Protoc 1560:313–330. https://doi.org/10.1007/978-1-4939-6788-9_24
Dhifi W, Bellili S, Jazi S, Bahloul N, Mnif W (2016) Essential oils chemical characterization and investigation of some biological activities: a critical review. Medicines 3(4):25
Dmitruk M, Sulborska A, Żuraw B, Stawiarz E, Weryszko-Chmielewska E (2019) Sites of secretion of bioactive compounds in leaves of Dracocephalum moldavica L.: anatomical, histochemical, and essential oil study. Braz J Bot 42:701–715. https://doi.org/10.1007/s40415-019-00559-6
Fahn A (1988) Tansley review no. 14: secretory tissues in vascular plants. New Phytol 108(3):229–257. https://doi.org/10.1111/j.1469-8137.1988.tb04159.x
Gontar L, Herman A, Osinska E (2021) Monarda essential oils as natural cosmetic preservative systems. Nat Volatiles Essent Oils 8(1):29–38
Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z (2011) Full-length transcriptome assembly from RNA-seq data without a reference genome. Nat Biotechnol 29(7):644–652. https://doi.org/10.1038/nbt.1883
Grzeszczuk M, Wesołowska A, Stefaniak A (2020) Biological value and essential oil composition of two Monarda species flowers. Acta Scientiarum Polonorum Hortorum Cultus 19(4):105–119. https://doi.org/10.24326/asphc.2020.4.10
Haratym W, Weryszko-Chmielewska E (2017) Ultrastructural and histochemical analysis of glandular trichomes of Marrubium vulgare L. (Lamiaceae). Flora 231:11–20. https://doi.org/10.1016/J.FLORA.2017.04.001
Harkat-Madouri L, Asma B, Madani K, Said ZBOS, Rigou P, Grenier D, Boulekbache-Makhlouf L (2015) Chemical composition, antibacterial and antioxidant activities of essential oil of Eucalyptus globulus from Algeria. Ind Crops Prod 78:148–153. https://doi.org/10.1016/J.INDCROP.2015.10.015
Kalicharan B, Naidoo Y, Heneidak S, Bhatt A (2015) Distribution, morphological and histochemical characteristics of foliar trichomes of Plectranthus zuluensis (Lamiaceae). Brazilian J Bot 38:961–971. https://doi.org/10.1007/s40415-015-0194-2
Katoch M, Pull S (2017) Endophytic fungi associated with Monarda citriodora, an aromatic and medicinal plant and their biocontrol potential. Pharm Biol 55(1):1528–1535. https://doi.org/10.1080/13880209.2017.1309054
Konarska A, Weryszko-Chmielewska E, Matysik-Woźniak A, Sulborska A, Polak B, Dmitruk M, Piotrowska-Weryszko K, Stefańczyk B, Rejdak R (2021) Histochemical and phytochemical analysis of Lamium album subsp. album l. corolla: essential oil triterpenes and iridoids. Molecules 26(14):4166. https://doi.org/10.3390/molecules26144166
Krause ST, Liao P, Crocoll C, Boachon B, Förster C, Leidecker F, Wiese N, Zhao D, Wood JC, Buell CR, Gershenzon J (2021) The biosynthesis of thymol, carvacrol, and thymohydroquinone in Lamiaceae proceeds via cytochrome P450s and a short-chain dehydrogenase. Proc Natl Acad Sci USA 118(52):e2110092118. https://doi.org/10.1073/pnas.2110092118
Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10(3):1–10. https://doi.org/10.1186/gb-2009-10-3-r25
Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform 12:1–16. https://doi.org/10.1186/1471-2105-12-323
Li W, Godzik A (2006) Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics 22:1658–1659. https://doi.org/10.1093/bioinformatics/btl158
Liao P, Zhou W, Zhang L, Wang J, Yan X, Zhang Y, Zhang R, Li L, Zhou G, Kai G (2009) Molecular cloning, characterization and expression analysis of a new gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from Salvia miltiorrhiza. Acta Physiol Plant 31:565–572. https://doi.org/10.1007/s11738-008-0266-z
Liu MQ, Liu ZW, Zhou J (2012) Morphology and histochemistry of the glandular trichomes of Isodon rubescens (Hemsley) H. Hara [Lamiaceae]: a promising medicinal plant of China. J Med Plant Res 6(8):1455–1460
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408. https://doi.org/10.1006/METH.2001.1262
Lohse M, Nagel A, Herter T, May P, Schroda M, Zrenner R, Tohge T, Fernie AR, Stitt M, Usadel B (2014) Mercator : a fast and simple web server for genome scale functional annotation of plant sequence data. Plant Cell Environ 37:1250–1258. https://doi.org/10.1111/pce.12231
Majdi M, Malekzadeh-Mashhady A, Maroufi A, Crocoll C (2017) Tissue-specific gene-expression patterns of genes associated with thymol/carvacrol biosynthesis in thyme (Thymus vulgaris L.) and their differential changes upon treatment with abiotic elicitors. Plant Physiol Biochem 115:152–162. https://doi.org/10.1016/J.PLAPHY.2017.03.016
Mannethody S, Sunojkumar P (2018) Trichome micromorphology and its systematic significance in Asian leucas (Lamiaceae). Flora 242:70–78. https://doi.org/10.1016/J.FLORA.2018.03.007
Masyita A, Sari RM, Astuti AD, Yasir B, Rumata NR, Emran TB, Nainu F, Simal-Gandara J (2022) Terpenes and terpenoids as main bioactive compounds of essential oils, their roles in human health and potential application as natural food preservatives. Food Chem X 13:117. https://doi.org/10.1016/J.FOCHX.2022.100217
Maurya S, Chandra M, Yadav RK, Narnoliya LK, Sangwan RS, Bansal S, Sandhu P, Singh U, Kumar D, Sangwan NS (2019) Interspecies comparative features of trichomes in Ocimum reveal insights for biosynthesis of specialized essential oil metabolites. Protoplasma 256:893–907. https://doi.org/10.1007/s00709-018-01338-y
Mishra A, Gupta P, Lal RK, Dhawan SS (2021) Assessing and integrating the transcriptome analysis with plant development, trichomes, and secondary metabolites yield potential in Mentha arvensisL. Plant Physiol Biochem 162:517–530. https://doi.org/10.1016/J.PLAPHY.2021.03.009
Misra P, Pandey A, Tiwari M, Chandrashekar K, Sidhu OP, Asif MH, Chakrabarty D, Singh PK, Trivedi PK, Nath P, Tuli R (2010) Modulation of transcriptome and metabolome of tobacco by Arabidopsis transcription factor, AtMYB12, leads to insect resistance. Plant Physiol 152(4):2258–2268. https://doi.org/10.1104/pp.109.150979
Moriya Y, Itoh M, Okuda S, Yoshizawa AC, Kanehisa M (2007) KAAS: An automatic genome annotation and pathway reconstruction server. Nucleic Acids Res 35:182–185. https://doi.org/10.1093/nar/gkm321
Nautiyal AK, Gani U, Sharma P, Kundan M, Fayaz M, Lattoo SK, Misra P (2020) Comprehensive transcriptome analysis provides insights into metabolic and gene regulatory networks in trichomes of Nicotiana tabacum. Plant Mol Biol 102:625–644. https://doi.org/10.1007/s11103-020-00968-2
Nazar N, Howard C, Slater A, Sgamma T (2022) Challenges in medicinal and aromatic plants DNA barcoding—lessons from the Lamiaceae. Plants 11(1):137. https://doi.org/10.3390/plants11010137
Padmanaban A, Salowsky R, Cher C (2012) RNA quality control using the agilent 2200 TapeStation system–assessment of the RIN e quality metric. Agilent technologies application notes. https://hpst.cz/sites/default/files/oldfiles/rna-quality-control-using-agilent-2200-tapestation-system-assessment-rine-quality-metric.pdf
Pathania AS, Guru SK, Verma MK, Sharma C, Abdullah ST, Malik F, Chandra S, Katoch M, Bhushan S (2013) Disruption of the PI3K/AKT/mTOR signaling cascade and induction of apoptosis in HL-60 cells by an essential oil from Monarda citriodora. Food Chem Toxicol 62:246–254. https://doi.org/10.1016/J.FCT.2013.08.037
Qi X, Chen Z, Yu X, Li L, Bai Y, Fang H, Liang C (2022) Characterisation of the Mentha canadensis R2R3-MYB transcription factor gene McMIXTA and its involvement in peltate glandular trichome development. BMC Plant Biol 22(1):219. https://doi.org/10.1186/s12870-022-03614-9
Ramos da Silva LR, Ferreira OO, Cruz JN, de Jesus Pereira FC, Oliveira dos Anjos T, Cascaes MM, Almeida da Costa W, Helena de Aguiar Andrade E, Santana de Oliveira M (2021) Lamiaceae essential oils, phytochemical profile, antioxidant, and biological activities. Evid Based Compl Altern Med. https://doi.org/10.1155/2021/6748052
Rather GA, Sharma A, Jeelani SM, Misra P, Kaul V, Lattoo SK (2019) Metabolic and transcriptional analyses in response to potent inhibitors establish MEP pathway as major route for camptothecin biosynthesis in Nothapodytes nimmoniana (Graham) Mabb. BMC Plant Biol 19:1–15. https://doi.org/10.1186/s12870-019-1912-x
Rehman R, Hanif MA, Mushtaq Z, Al-Sadi AM (2016) Biosynthesis of essential oils in aromatic plants: a review. Food Rev Int 32(2):117–160. https://doi.org/10.1080/87559129.2015.1057841
Rodrigues L, Póvoa O, Teixeira G, Figueiredo AC, Moldão M, Monteiro A (2013) Trichomes micromorphology and essential oil variation at different developmental stages of cultivated and wild growing Mentha pulegium L. populations from Portugal. Ind Crops Prod 43:692–700. https://doi.org/10.1016/J.INDCROP.2012.07.061
Sarker LS, Mahmoud SS (2015) Cloning and functional characterization of two monoterpene acetyltransferases from glandular trichomes of L. x intermedia. Planta 242:709–719. https://doi.org/10.1007/s00425-015-2325-1
Sarker LS, Adal AM, Mahmoud SS (2020) Diverse transcription factors control monoterpene synthase expression in lavender (Lavandula). Planta 251:5. https://doi.org/10.1007/s00425-019-03298-w
Schuurink R, Tissier A (2020) Glandular trichomes: micro-organs with model status? New Phytol 225(6):2251–2266. https://doi.org/10.1111/nph.16283
Shah M, Alharby HF, Hakeem KR, Ali N, Rahman IU, Munawar M, Anwar Y (2020) De novo transcriptome analysis of Lantana camara L. revealed candidate genes involved in phenylpropanoid biosynthesis pathway. Sci Rep 10(1):13726. https://doi.org/10.1038/s41598-020-70635-5
Sharifi-Rad J, Sureda A, Tenore GC, Daglia M, Sharifi-Rad M, Valussi M, Tundis R, Sharifi-Rad M, Loizzo MR, Ademiluyi AO, Sharifi-Rad R (2017) Biological activities of essential oils: From plant chemoecology to traditional healing systems. Molecules 22(1):70. https://doi.org/10.3390/molecules22010070
Sharma S, Sangwan NS, Sangwan RS (2003) Developmental process of essential oil glandular trichome collapsing in menthol mint. Curr Sci 84:544–550
Simão FA, Waterhouse RM, Ioannidis P, Kriventseva EV, Zdobnov EM (2015) BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics 31(19):3210–3212. https://doi.org/10.1093/bioinformatics/btv351
Tabanca N, Bernier UR, Ali A, Wang M, Demirci B, Blyth EK, Khan SI, Baser KHC, Khan IA (2013) Bioassay-guided investigation of two Monarda essential oils as repellents of yellow fever mosquito Aedes aegypti. J Agric Food Chem 61(36):8573–8580. https://doi.org/10.1021/jf402182h
Tan H, Xiao L, Gao S, Li Q, Chen J, Xiao Y, Ji Q, Chen R, Chen W, Zhang L (2015) TRICHOME AND ARTEMISININ REGULATOR 1 is required for trichome development and artemisinin biosynthesis in Artemisia annua. Mol Plant 8(9):1396–1411. https://doi.org/10.1016/j.molp.2015.04.002
Tang HM, Jiang Q, Liu HY, Zhang F, Liu Q, Pu GB, Li J, Wang LN, Zhang YQ (2022) Glandular trichomes of medicinal plants: types, separation and purification, biological activities. Biol Plant 66:219–227. https://doi.org/10.32615/bp.2022.027
Turner GW, Gershenzon J, Croteau RB (2000) Distribution of peltate glandular trichomes on developing leaves of peppermint. Plant Physiol 124(2):655–664. https://doi.org/10.1104/pp.124.2.655
Wagner GJ, Wang E, Shepherd RW (2004) New approaches for studying and exploiting an old protuberance, the plant trichome. Ann Bot 93:1–3. https://doi.org/10.1093/aob/mch011
Walter MH, Hans J, Strack D (2002) Two distantly related genes encoding 1-deoxy-D-xylulose 5-phosphate synthases: differential regulation in shoots and apocarotenoid-accumulating mycorrhizal roots. Plant J 31(3):243–254. https://doi.org/10.1046/j.1365-313X.2002.01352.x
Wang M, Gao M, Zhao Y, Chen Y, Wu L, Yin H, Yang J, Xiong S, Wang S, Wang J, Yang Y (2022) LcERF19, an AP2/ERF transcription factor from Litsea cubeba, positively regulates geranial and neral biosynthesis. Hortic Res 9:euhac093. https://doi.org/10.1093/hr/uhac093
Werker E, Ravid U, Putievsky E (1985) Glandular hairs and their secretions in the vegetative and reproductive organs of Salvia sclarea and S. dominica. Isr J Plant Sci 34(2–4):239–252
Wu M, Chang J, Han X, Shen J, Yang L, Hu S, Huang BB, Xu H, Xu M, Wu S, Li P (2023) A HD-ZIP transcription factor specifies fates of multicellular trichomes via dosage-dependent mechanisms in tomato. Dev Cell 58(4):278–288. https://doi.org/10.1016/j.devcel.2023.01.009
Xu J, van Herwijnen ZO, Dräger DB, Sui C, Haring MA, Schuurink RC (2018) SlMYC1 regulates type VI glandular trichome formation and terpene biosynthesis in tomato glandular cells. Plant Cell 30(12):2988–3005. https://doi.org/10.1105/tpc.18.00571
Yan T, Chen M, Shen Q, Li L, Fu X, Pan Q, Tang Y, Shi P, Lv Z, Jiang W, Ma YN (2017) HOMEODOMAIN PROTEIN 1 is required for jasmonate-mediated glandular trichome initiation in Artemisia annua. New Phytol 213(3):1145–1155. https://doi.org/10.1111/nph.14205
Yan X, Li W, Liang D, Zhao G, Caiyin Q, Qiao J (2021) Comparative transcriptome analysis of sesquiterpene biosynthesis and functional characterization of sesquiterpene synthases in Leonurus sibiricus L. Planta 253:71. https://doi.org/10.1007/s00425-021-03586-4
Yang C, Marillonnet S, Tissier A (2021) The scarecrow-like transcription factor SlSCL3 regulates volatile terpene biosynthesis and glandular trichome size in tomato (Solanum lycopersicum). Plant J 107(4):1102–1118. https://doi.org/10.1111/tpj.15371
Zhou F, Pichersky E (2020) More is better: the diversity of terpene metabolism in plants. Curr Opin Plant Biol 55:1–10. https://doi.org/10.1016/j.pbi.2020.01.005
Acknowledgements
PM acknowledges the funding support by Council of Scientific and Industrial Research (CSIR), Government of India in the form of the project CSIR Aroma Mission Phase-II (HCP-007). MAW and MF acknowledge UGC, New Delhi, India and CSIR, India for junior and senior research fellowships. The manuscript has institutional publication number CSIR-IIIM/IPR/00564.
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Sharma, P., Wajid, M.A., Fayaz, M. et al. Morphological, phytochemical, and transcriptome analyses provide insights into the biosynthesis of monoterpenes in Monarda citriodora. Planta 258, 49 (2023). https://doi.org/10.1007/s00425-023-04207-y
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DOI: https://doi.org/10.1007/s00425-023-04207-y