Gas chromatography-mass spectrometry in the taxonomy of Miscanthus

Chemotaxonomy as a system approach deals with intra- and interspecific polymorphism of a group of taxa in order to clarify their taxonomic positions or to select material for selection or introduction. In this study we performed chemotaxonomic analysis of specimens of Miscanthus sinensis and M. sacchariflorus collected in the Russian Far East and of hybrid plants of both natural and artificial origin. We found 153 substances and identified 143 of them in extracts of eleven Miscanthus plants by gas chromatography-mass spectrometry (GC-MS). These substances can be grouped into alkanes (20 compounds), fatty acids (34), phenols (13), sterols (18) toсopherols (8), norterpenoids (12), and phytols (13), as well as their derivatives. The main components of the extracts of miscanthus samples are fatty acids and their derivatives (total content 19.94–41.02 %), dominated by palmitic and linolenic acids, and sterols (mainly β-sitosterol, stigmasterol, and α-amyrin), which constitute 17.15–31.73 %. The values of the CPI “oddness index” for the alkane components of the extracts were within 1.55–7.18, with extracts from leaves of the Far Eastern samples characterized by the lower half of this range (1.55–2.74), while extracts from leaves of hybrids fell to the upper half (5.78–7.18). Principal component analysis of extraction profiles allowed us to separate three distinct clusters: M. sinensis, M. sachariflorus, and their hybrids, as well as to verify the origin of one of the natural hybrids. The results of chemotaxonomic analysis mostly matched those of DNA sequencing of a fragment of the plastid genome, which, moreover, allowed us to identify the species nature of the maternal plants used to obtain these hybrids. Chemotaxonomic analysis using GC-MS was found to be an efficient additional technique to delimit various morphological forms of M. sinensis, M. sachariflorus, and their hybrids.

1. Darmogray S.V., Erofeeva N.S., Filippova A.S., Darmogray V.N. Chemotaxonomical study of the Cerastium genus some plants of the Caryophyllaceae family. Uspekhi Sovremennogo Yestestvoznaniya = Advances in Current Natural Sciences. 2016;7:32-36. (in Russian)

2. Pokrovskaya I.S., Mazova O.V., Apykhtin N.N., Plemenkov V.V. Chemotaxonomy yarrow (Achillea millefolium L.). Khimiya Rastitelnogo Syr’ya = Chemistry of Plant Raw Materials. 2009;3:85-89. (in Russian)

3. Buckingham J. Dictionary of Organic Compounds. CRC Press, 1996; 8:3681.

4. Chen S.L., Renvoize S.A. Miscanthus. In: Wu Z.Y., Raven P.H., Hong D.Y. (Ed.). Flora of China. Beijing/St. Louis: Science Press/ Missouri Botanical Garden Press, 2006;22:581-583.

5. Dool H.V.D., Kratz P.D., A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography. J. Chromatogr. A. 1963;11:463-471. DOI 10.1016/S0021-9673(01)80947-X.

6. Eglinton G., Hamilton R.J. Leaf epicuticular waxes. Science. 1967; 156:1322-1335.

7. Hammer Ø., Harper D.A.T., Ryan P.D. Past: Paleontological Statistics Software Package for Education and Data Analysis. Palaeontologia Electronica. 2001;4(1):1-9. 178kb. http://palaeo-electronica.org/2001_1/past/issue1_01.htm.

8. Hodkinson T.R., Petrunenko E., Klaas M., Münnich C., Barth S., Shekhovtsov S.V., Peltek S.E. New breeding collections of Miscanthus sinensis, M. sacchariflorus and hybrids from Primorsky Krai, Far Eastern Russia. In: Perennial Biomass Crops for a Resource-Constrained World. Springer, 2016;105-118.

9. Hodkinson T.R., Renvoize S.A. Nomenclature of Miscanthus × giganteus (Poaceae). Kew Bull. 2001;56:759-760. DOI 10.2307/4117709.

10. Jetter R., Kunst L., Samuels A.L. Composition of plant cuticular waxes. In: Riederer M., Muller C. (Ed). Biology of the Plant Cuticle. Oxford: Blackwell Publ., 2006.

11. Linde-Laursen I.B. Cytogenetic analysis of Miscanthus ‘Giganteus’, an interspecific hybrid. Hereditas. 1993;119:297-300. DOI 10.1111/j.1601-5223.1993.00297.x.

12. Namzalov B.B., Zhigzhitzhapova S.V., Dubrovsky N.G., Sakhyaeva A.B., Radnaeva L.D. Wormwoods of Buryatia: diversity analysis, ecological-geographical features, and chemotaxonomy of section Abrotanum. Acta Biologica Sibirica. 2019;5(3):178-187. DOI 10.14258/abs.v5.i3.6589.

13. Osorio C., Duque C., Batista-Viera F. Studies on aroma generation in lulo (Solanum quitoense): enzymatic hydrolysis of glycosides from leaves. Food Chem. 2003;81(3):333-340. DOI 10.1016/S0308-8146(02)00427-2.

14. Shekhovtsov S.V., Shekhovtsova I.N., Peltek S.E.. Phylogeny of Siberian species of Carex sect. Vesicariae based on nuclear and plastid markers. Nord. J. Bot. 2012;30(3):343-351. DOI 10.1111/j.1756-1051.2011.01405.x.

15. Silva F.M.A., Silva F.F.A., Lima B.R., Almeida R.A., Soares E.R., Koolen H.H.F., Souza A.D.L., Pinheiro M.L.B. Chemotaxonomy of the Amazonian Unonopsis species based on leaf alkaloid fingerprint direct infusion ESI-MS and chemometric analysis. J. Braz. Chem. Soc. 2016;27:599-604. DOI 10.5935/0103-5053.20150296.

16. Socaci S.A., Socaciu C., Tofană M., Raţi I., Pinteaa A. In-tube extraction and GC-MS analysis of volatile components from wild and cultivated sea buckthorn (Hippophae rhamnoides L. ssp. Carpatica) berry varieties and juice. Phytochem. Anal. 2013;24:319-328. DOI 10.1002/pca.2413. Epub 2013 Jan 15.

17. Villaverde J.J., Domingues R.M.A., Freire C.S.R., Silvestre A.J.D., Pascoal Neto C., Ligero P., Vega A. Miscanthus × giganteus extractives: a source of valuable phenolic compounds and sterols. J. Agric. Food Chem. 2009;57:3626-3631. DOI 10.1021/jf900071t.

18. Weißmann G.V., Lange W. Untersuchung der Extraktstoffe von Miscanthus sinensis Anderss. Holzforschung. 1991;45(4):285-289.

19. Winkler-Moser J. Gas Chromatographic Analysis of Plant Sterols. USDA, ARS, NCAUR, Functional Foods Research Unit, 2012. DOI 10.21748/lipidlibrary.40384.

20. Yi Duan, Jinxian He. Distribution and isotopic composition of n-alkanes from grass, reed and tree leaves along a latitudinal gradient in China. Geochem. J. 2011;45:199-207. DOI 10.2343/geochemj.1.0115.