Morphological, Molecular and Phytochemical Variation in Some Thyme Genotypes

Authors

1 East and North-East Region Agricultural Biotechnology Research Institute, Mashhad, Iran.

2 Khorasan Razavi Agricultural and Natural Resources Research Center, Education and Extension Organization (AREEO), Mashhad, Iran

Abstract

Thyme is an important medicinal plant in cosmetic, pharmaceutical and food industries. The first step for breeding of thyme is evaluating of genetic variation and relationship between thyme’s accessions. Therefore, the objective of this study was to evaluate morphology, chemical and molecular variation of 13 accessions of Thyme medicinal plant. ANOVA showed significant differences between accessions for total characterization tested. The dendrogram constructed on the basis of morphology similarities showed two major clusters. In order to evaluate the genetic variation, the genomic DNA was extracted using modified medicinal CTAB protocol. The evaluation of the of DNA quality was performed using electrophoresis. Twenty primers were used for PCR analysis and only 9 primers showed clear bands. Out of 149 bands, 83.22% were the polymorphism. The data were analyzed with SPSS and POPGENE programs and the dendrogram was drawn based on UPGMA and showed three major clusters. In order to evaluate the chemical variation, essential oil was obtained using Clevenger unit. ANOVA showed significant differences between accessions for total characterization test. Dendrogram for chemical variation showed two major clusters.Chemical and morphological traits’ matrices were formed using Statistical V5.5A software and were compared with genetic similarity matrices using GenAlex 6.1 software.

References

1. Morales R. The history, botany and taxonomy of genus Thymus Elisabeth Stahl- Biskup and Saez, F.(eds). The genus Thymus. 2002;1-124. 
2. Omid Baigy R. Production and processing of medicinal plants.  Second Volume. Astan Qods Razavi publications. 2007; 280 pages. (In Persian).
3. Stahl- Biskup E, Saez  F. Thyme.1st ed. Talor and Francis. England, 2002. 346 pages.
4. Rahimmalek M, Bahreininejad B, Khorrami M, Tabatabaei S.B.E. Genetic variability and geographic differentiation in Thymus daenensis subsp. daenensis, an endangered medicinal plant, as revealed by inter-simple sequence repeat (ISSR) markers. Biochem Genet. 2009;47:831-842.
5. Amin G. Popular medicinal plants of Iran. Tehran University of Medical Sciences Press. Tehran. 2005. (In Persian).
6. GhasemiPirbalouti A, Karimi A, Yousefi M, Enteshari S, Golparvar AR. Diversity of Thymus daenensis Celak in central and west of Iran. J Med Plants Res. 2011;5:319-323.
7. Khoshsokhan F, Babalar M, Fatahi M, Poormeidani A. Assessment of Genetic Diversity of Some Wild Populations of Thymus Kotschyanus using RAPD Molecular Markers. Cercetari Agronomice in Moldova. 2014;3:71-81.
8. Alamdary S.B.L, Safarnejad A, Rezaee M. Evaluation of genetic variation between Thymus accessions using molecular markers. J Basic Appl Sci Res. 2011;1:2552-2556.
9. Alamdary S.B.L, Safarnejad A, Nematzadeh G.A. Using RAPD marker for genetic diversity assessment of several Thymus species. Iranian J Rangelands Forests Plant Breeding Gen Res. 2012; 20:192-201.
10. Bagherzadeh F. Genetic diversity assessment of thyme and evaluation of relationship using RAPD marker. MSc thesis. Ferdowsi University. Mashhad. Iran. 2009. (In Persian).
11. Echeverrigaray S, Agostini G, Atti-Serfini L, Paroul N, Pauletti G. F, Atti Dos Santos A.C. Correlation between the chemical and genetic relationships among commercial thyme cultivars. J Agric Food Chem. 2001;49:4220-422.
12. Trindade H,  Costa M,  Lima A. S, Pedro L .G, Ana C, FigueiredoJ, Barroso G. Genetic diversity and chemical polymorphism of Thymus caespititius from Pico, São Jorge and Terceira islands (Azores), Biochem System Ecol. 2008;36:790-797.
13. Trindade H,  Costa M,  Lima AS, Pedro LG, Ana C, FigueiredoJ, Barroso G. A combined approach using RAPD, ISSR and volatile analysis for the characterization of Thymus caespititius from Flores, Corvo and Graciosa islands (Azores, Portugal). Biochem Syst Ecol. 2009;37:670-677.  
14. Daftari Z, Safarnejad A. Karyotypic study of four Thymus species (T. pubescens, T. fedtschenkoi, T. daenensis, T. lancifolius). Iranian J Rangelands Forests Plant Breeding Gen Res. 2012;19:241-250.
15. Zia A, Rezanejad F, Safarnejad A. In vitro selection for NaCl tolerance in Thymus vulgaris L. J Cell Mol Res. 2011;2:86-92.
16. Fracaro F, Echeverrigaray S. Genetic variability in Hesperozygis ringens Benth. (Lamiaceae), an endangered aromatic and medicinal plant of Southern Brazil. Biochem Genet. 2006;44:479-490.
17. Dyke F.V. Conservation biology, foundations, concepts. applications. 2008. Springer. 
18. Ben el Hadj Ali I, Guetat A, Boussaid M. Chemical and genetic variability of Thymus algeriensis Bioss. et Reut. (Lamiaceae), a North African species. Indust Crops Prod. 2012;40:277-284.
19. Thompson JD. Population  structure  and the spatial dynamics of genetic polymorphism in thyme  in Stahl – Biskup E. & Saez, F.(eds.) The genus Thymus. 2002;76 -122.
20. Nematzadeh Gh, Kiani GH. Plant breeding book (classic methods). First Vollum. Mazandaran University publications. 2005. 456 pages. (In Persian).
21. Hashemi H, Safarnejad A, Bagheri A. Investigation of genetic variation among Iran’s Persian Zira (Bunium persicum Boiss) landraces using RAPD marker. Iranian J Rangelands Forests Plant Breeding Genetic Research. 2009;16:239-246.
22. Farsi M, Zolala J. Principles of Plant Biotechnology (Translation). The Ferdowsi University of Mashhad publications. Second edition. 2004. 495 pages.
23. Akbarinia A, Sharifi Ashoorabadi A, Mirza M. Evaluation of performance, value and principalcompounds in the essential oil of (Thymus daenensis Celak.) cultivated in Qazvin. Investigation Med Arom Plants. 2010;26:205-212. (In Persian).
24. Rezaie M.B, Jaymand K. Study of chemical constituent oil of Lippia citrodora H.B.K. Pajouhesh-va-Sazandegi in Agronomy and Horticulture. 2002;53:13-15. (In Persian).
25. Khanuja  S.P.S, Shasany  AK, Kumar S. RAPD isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Mol Biol Rep. 1999;17:1-7.
26. Ward J.H. Hierarchical grouping to optimize an objective function. J Am Stat Assoc. 1963;58:236-244.
27- Nei M. Estimation of average heterozygosity and genetic distance from a small number of individual. Genetics .1987;89:583-590.
28. Shannon CE. A mathematical theory of communication. Bell Syst Tech J. 1948;27:379-423 and 623-656.
29. Kermani M, Marashi H, Safarnejad A. Investigation of genetic variation within and among two species of Cuminum spp. using AFLP markers. Iranian J Rangelands & Forests Plant Breeding & Gen Res. 2009;16:198-206.
30. Khadivi-Khub A, Salehi-Arjmand H, Movahedi KH, Hadian J. Molecular and morphological variability of Satureja bachtiarica in Iran. PlantPlant Syst Evol. 2015;301:77–93.
31. Hadian J, Tabatabaei SMF, Naghavi MR, Jamzad Z, Ramak-Masoumi T. Genetic diversity of Iranian accessions of Satureja hortensis L. based on horticultural traits and RAPD markers. Scientia Horticulturae. 2008;115:196-202.
32. Jamzad Z. Thymus and Satureja species. Institue Research Forests and Rangeland. 2010. 171 pages. (In Persian).
33. Hashemi H, Safarnejad A, Bagheri A. The use of RAPD marker assessing the genetic diversity of Bunium persicum B. Fedtsch populations. Int J Sci Nat. 2010;1:202-208.
34. Safarnejad A, Khosravinasab Z. Cytogenetic study of Cuminum setifolium (Boiss) Kos-Pol as a medicinal plant. Int J Sci Nat. 2011;2:105-109.
35. Safarnejad A, Khosravinasab Z. Comparison of biochemical characterization in Cuminum cyminum and Cuminum setifolium seeds. Int J Sci Nat. 2011;2:874-877. 
36- Harrison RE, Luby JJ, Furnier GR, Hancock JF. Morphological and molecular variation among populations of octoploid Fragaria virginiana and chiloensis (Rosaceae) from North America. Am J Bot. 1997;84:612-620.                                     
37. Persson HA, Rumpunen K, Molersted LK. Identification of culinary rhubarb (Rheum ssp.) cultivars using morphological characterization and RAPD marker. J Hortic Sci Biotech. 2000;75: 684-689.
38. Samal S, Rout GR, Nayak S, Nanda RM, Lenka PC, Das S. Primer screening and optimization for RAPD analysis of cashew. J Plant Biol. 2003;46:301-304.
39. Lihová1 J, Kučera1 J, Perný1 M, Marhold K. Hybridization between two Polyploid Cardamine (Brassicaceae) Species in North-western Spain: Discordance Between Morphological and Genetic Variation Patterns. Ann Bot. 2007;99:1083-1096.
40. Khan MA, Witzke-Ehbrecht S, Maass BL, Becker HC. Relationships among different geographical groups, agro-morphology, fatty acid composition and RAPD marker diversity in safflower (Carthamus tinctorius). Genet. Resource. Crop Evolution. 2008;56:19-30.
41. Panahi B, Afzal R, Mahmoodnia M, Neghab MG. Relationships among AFLP, RAPD marker diversity and agro-morphology traits, in safflower (Carthamus tinctoriusL.). World Appl Sci J. 2013;26:213-223.
42. Persson HA, Gustavsson BA. The extent of clonality and genetic diversity in lingonberry (Vaccinium vitis-idaeaL.) revealed by RAPDs and leaf-shape analysis. Mol Ecol. 2001;10:1385-1397.
43. Stokl J, Schlueter PM, Stuessy TF, Paulus HF, Assum G, Ayasse M. Scent variation and hybridization cause the displacement of a sexually deceptive orchid species. Am J Bot. 2008;95:472-481.
44. Feulner M, Pointner S, Heuss L, Aas G, Paule J, Dotterl S. Floral scent and its correlation with AFLP data in Sorbus. Org Div Evol. 2014;14:339-348.
45. Lopez PA, Widrlechner MP, Simon PW, Rai S, Boylston TB, Isbell TA, Bailey TB, Gardner CA, Wilson LA. Assessing phenotypic, biochemical, and molecular diversity in coriander (Coriandrum sativumL.) germplasm. Genet Res Crop Evol. 2008;55:247-275.
46. Azizi A, Hadian J, Gholami M, Friedt W, Honermeier B. Correlations between Genetic, Morphological, and Chemical Diversities in a Germplasm Collection of the Medicinal Plant Origanum vulgare L. Chem Biodiver. 2012;9:2784-2801.
47. Keskitalo M, Pehu E, Simon J. Variation in volatile compounds from tansy (Tanacetum vulgare L.) related to genetic and morphological differences between genotypes. Biochem Syst Ecol. 2001;29:267-285.
48. Kremer D, Bolaric S, Ballian D, Bogunic F, Steševic D, Karlovic K. Morphological, genetic and phytochemical variation of the endemic Teucrium arduini L. (Lamiaceae). Phytochemistry. 2015;116:111-119.
49. Rustaiee A.R,  Yavari A.,  Nazeri V, Shokrpour M,  Sefidkon F,  Rasouli M. Genetic Diversity and Chemical Polymorphism of Some Thymus Species. Chem Biodiver. 2013;10:1088-1098.
50. Pluhár Z, Kocsis M, Kuczmog A, Csete S., Simkó H., Sárosi S., Molnár P., Horváth G. Essential oil composition and preliminary molecular study of four Hungarian Thymus species. Acta Bio Hungarica. 2012;63:81-96.
51. Labra M, Miele M, Ledda B, Grassi F, Mazzei M, Sala F. Morphological characterization, essential oil composition and DNA genotyping of Ocimum basilicum L. cultivars. Plant Sci. 2004;167:725-73.
Journal of Medicinal plants and By-Products
Volume 6, Issue 1
April 2017
Pages 41-52

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