Ethyl methane sulfonate (EMS) enhanced the formation of leaf glandular trichomes and the production of artemisinin in Artemisia annua L.
Article Sidebar
Main Article Content
Abstract
Artemisia annua L.can naturally synthesize artemisinin, an anti-malarial compound. Due to the naturally low content of artemisinin available in this plant, many different approaches were used to increase the production of artemisinin. The objective of our study was to determine whether ethyl methane sulfonate (EMS) treated seeds could enhance the formation of leaf glandular trichomes and increase the artemisinin content in the plants. The in vitro plantlets were established from EMS treated and untreated seeds on a basic Murashige and Skoog solid medium. The morphology of the leaf glandular trichomes was observed under a fluorescence microscope (Olympus FX53) at the blue wavelength (460-495 nm) while the artemisinin content was determined using High-performance liquid chromatography (HPLC). Results indicated that EMS did not affect the plantlet growth but caused temporary morphological abnormality in < 17% of the in vitro plantlets in the form of leaf albinism and chlorosis which disappeared after four subculture cycles. EMS of 0.5-1% (v/v) used for the seed treatment was able to induce the formation of more leaf glandular trichomes for all the three tested A. annua L. clones. While EMS ranging between 1.0% (v/v) and 2.0% (v/v) caused an increase in trichome size. A higher artemisinin content was detected in plantlets derived from seeds treated with 1.0% (v/v) EMS. Hence it was concluded that 1% (v/v) EMS could be used to induce the formation of more and bigger leaf trichomes that enhance the production of artemisinin in A. annua L.
Article Details
References
Saha P, Ganguly S, Dutta S, Kundu PK, Bera DK, Basu N, et al. Impact of artemisinin-based combination therapy on falciparum malaria in urban Kolkata: a clinic-based report. Jpn J Infect Dis. 2015;68(4):321-323.
Bui TTT, Minh TV, Boey PL, Chan LK. Effects of environmental factors on growth and artemisinin content of Artemista annua L. Trop Life Sci Res. 2011;22(2):37-43.
Davies MJ, Atkinson CJ, Burns C, Arroo R, Woolley J. Increases in leaf artemisinin concentration in Artemisia annua L. in response to the application of phosphorus and boron. Ind Crop Prod. 2011;34:1465-1473.
Brisibe EA, Udensi O, Chukwurah PN, de Magalhäes PM, Figueira GM, Ferreira JFS. Adaptation and agronomic performance of Artemisia annua L. under lowland humid tropical conditions. Ind Crop Prod. 2012;39(1):190-197.
Duke S, Paul R. Development and fine structure of the glandular trichomes of Artemisia annua L. Int J Plant Sci. 1993;154(1):107-118.
Duke MV, Paul RN, Elsohly HN, Sturtz G, Duke SO. Localization of artemisinin and artemisitene in foliar tissues of glanded and glandless biotypes of Artemisia annua L. Int J Plant Sci.1994; 155:365-372.
Tan H, Xiao L, Gao S, Li Q, Chen J, Xiao Y, et al. Trichome and artemisinin regulator 1 is required for trichome development and artemisinin biosynthesis in Artemisia annua L. Mol Plant. 2015; 8:1396-1411.
Kapoor R, Chaudhary V, Bhatnagar AK. Effects of arbuscular mycorrhiza and phosphorus application on artemisinin concentration in Artemisia annua L. Mycorrhiza. 2007;17(7):581-587.
Olsson ME, Olofsson LM, Lindahl AL, Lundgren A, Brodelius M, Brodelius PE. Localization of enzymes of artemisinin biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry. 2009;70(9):1123-1128.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol plantarum. 1962; 15:473-497.
Lapkin AA, Walker A, Sullivan N, Khambay B, Mlambo B, Chemat S. Development of HPLC analytical protocols for quantification of artemisinin in biomass and extracts. J Pharm Biomed Anal. 2009;49(4):908-915.
Caldwell DG, Mccallum N, Shaw P, Muehlbauer GJ, Marshall DF, Waugh R. A structured mutant population for forward and reverse genetics in Barley (Hordeum vulgare L.). Plant J. 2004;40(1):143-150.
Shah S, Gong ZH, Arisha MH, Khan A, Tian SL. Effect of ethyl methyl sulfonate concentration and different treatment conditions on germination and seedling growth of the cucumber cultivar Chinese long (9930). Genet Mol Res. 2015;14(1):2440-2449.
Gnanamurthy S, Dhanavel D. Effect of EMS on induced morphological mutants and chromosomal variation in cowpea (Vigna unguiculata (L.) Walp). Int Lett Nat Sci. 2014;22:33-43.
Wu JL, Wu C, Lei C, Baraoidan M, Bordeos A, Madamba MRS, et al. Chemical- and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics. Plant Mol Biol. 2005;59(1):85-97.
Feiz L, Beecher BS, Martin JM, Giroux MJ. In planta mutagenesis determines the functional regions of the wheat puroindoline proteins. Genetics. 2009;183(3):853-860.
Nehnevajova E, Herzig R, Federer G, Erismann KH, Schwitzgúebel JP. Chemical mutagenesis-A promising technique to increase metal concentration and extraction in sunflowers. Int J Phytoremediation. 2007;9(2):149-165.
Khursheed S, Khan S. Cytology of morphological mutants of Vicia faba L. var. vikrant. Annu Res Rev Biol. 2015;5(4):366-371.
Umavathi S, Mullainathan L. Mutagenic effect of gamma rays and EMS on seed germination, seedling height reduction and survivability of chick pea (Cicer arietinum L.) Var. Co-4. Int Lett Nat Sci. 2014;16:38- 43.
Kashid NG, Salve US. Effect of induced mutation on leaf morphological changes in okra (Abelmoschus esculentus L. Moench). Int J Recent Ad Muts Res. 2014;1(1):1-2.
Talebi AB, Talebi AB, Shahrokhifar B. Ethyl methane sulphonate (EMS) induced mutagenesis in Malaysian rice (cv. MR219) for lethal dose determination. Am J Plant Sci. 2012;3:1661-1665.
Bhosale RS, More AD. Effect of gamma radiation on seed germination, seedling height and seedling injury in Withania somnifera, (L.) Dunal. Int j plant sci. 2014;2(3):223-225.
Murthy VNY, Ramesh HL, Munirajappa. Ethyl methane sulphonate induced morphological variations in mulberry. J Appl Nat Sci. 2011;3(1):114-118.
Dhulgande GS, Dhale DA, Pachkore GL, Satpute RA. Mutagenic effectiveness and efficiency of gamma rays and ethyl methane sulphonate in pea (Pisum sativum L.). J Exp Sci. 2011;2(3):7-8.
Kumar G, Yadav RS. EMS induced genomic disorders in sesame (Sesamum indicum L.). Rom J Boil-Plant Boil. 2010;55(2):97-104.
Bossinger G, Spokevicius AV. Plant chimaeras and mosaic. John Wiley & Sons Ltd., Chichester, 2011.
Dalin P, Agren J, Bjorkman C, Huttunen P, Karkkainen K. Leaf trichome formation and plant resistance to herbivory. In: Schaller A., editor. Induced plant resistance to herbivory. Santa Barbara CA93106-6150, USA: University of California; 2008. p. 89-105.
Baur R, Binder S, Benz G. Nonglandular leaf trichomes as short-term inducible defense of the grey alder, Alnus incana (L.), against the chrysomelid beetle, Agelastica alni L. Oecologia. 1991;87(2):219-226.
Rautio P, Markkola A, Martel J, Tuomi J, Harma E, Kuikka K, et al. Developmental plasticity in brich leaves: defoliation causes a shift from glandular to non-glandular trichomes. Oikos. 2002;98:437-446.