Characterization of Morphological, Phytochemical and Molecular Diversity of Artemisia annua Accessions in Hyrcanian Area of Iran

Authors
M. R. Naghavi 1
E. Aryakia 2
S. Hadi 3
H. Ghafoori 4
H. Mousavi 2
H. Ramazani 2
M. Feyzbakhsh 2
Y. Ajani 2
Z. Farahmand 2
L. Poorhosseini 3
S. A. Shahzadeh Fazeli 2
1 Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Islamic Republic of Iran.
2 Iranian Biological Resource Center (IBRC), (ACECR), Karaj, Islamic Republic of Iran.
3 Department of Horticulture, Biotechnology and Molecular Genetics, College of Agriculture and Natural Resources, Karaj Islamic Azad University, Alborz, Islamic Republic of Iran.
4 Department of Biology, University of Guilan, Rasht, Islamic Republic of Iran.
Abstract
Artemisia annua L. is an important medicinal plant used as an original source of artemisinin for treating malaria. Although there is a wide distribution of A. annua in Hyrcanian Areas (Mazandaran, Guilan, and Golestan Provinces in N Iran), no considerable effort has been made for diversity assessment. In this study, morphological, phytochemical, and molecular characterization of A. annua accessions in this area were assessed using 4 quantitative characteristics (height, fresh weight, dry weight, and trichome density), artemisinin content, and 15 ISSR primers. Using these traits, a high level of morphological, phytochemical and molecular diversity was revealed among A. annua accessions in the provinces (populations) of Hyrcanian Areas. At inter-population level, the highest value of artemisinin was observed in Mazandaran Province. Moreover, significant correlation between artemisinin content and trichome density was observed that could be useful for indirect selection of artemisinin yield in different accessions of A. annua. In addition, fifteen ISSR primers generated a total number of 222 amplified bands, consisting of 177 and 45 polymorphic and monomorphic loci across the 60 accessions, respectively. These results indicate that ISSR-PCR is a reliable tool for fingerprinting A. annua at the intra-population level. Our results altogether are valid contributions for gene bank management and Artemisia annua breeding programs.

Keywords

Subjects

1. Agren, J. and Schemske, D. W. 1994. Evolution of Trichome Number in a Naturalized Population of Brassica rapa. Am. Nat., 143: 1–13.
2. Akhani, H., Djamali, M., Ghorbanalizadeh, A. and Ramezani, E. 2010. Plant Biodiversity of Hyrcanian Relict Forests, N Iran: An Overview of the Flora, Vegetation, Palaeoecology and Conservation. Pak. J. Bot., 42: 231-258.
3. Aryakia, E., Karimi, H., Naghavi, M. R. and Shahzadeh Fazeli, S. A. H. 2016. Morphological Characterization of Intra- and Inter-Specific Diversity in Some Iranian Wild Allium Species. Euphytica, 211:185-200.
4. Aryakia, E., Karimi, H., Naghavi, M. R., Yazdanfar, N. and Shahzadeh Fazeli, S. A. H. 2017. Evaluating Essential Oil Constituent of Four Allium Species of the Subgenus and Section Allium. Iran. J. Field Crop Sci., In press.
5. Aryakia, E., Naghavi, M. R., Farahmand, Z. and Shahzadeh Fazeli, S. A. H. 2015. Evaluating Allelopathic Effects of Some Plant Species in Tissue Culture Media as an Accurate Method for Selection of Tolerant Plant and Screening of Bioherbicides. J. Agr. Sci. Tech., 17: 1011–1023.
6. Aryakia, E., Ramazani, H., Ghafoori, H., Dolatyari, A., Naghavi, M. R. and Shahzadeh Fazeli, S. A. H. 2012. The Effect of Cryopreservation on Germination and Growth Indices of Some Orthodox Seeds. Iran J. Rangel. Plant Breed. Genet. Res. 19: 218–230.
7. Baghalian, K., Ziai, S. A., Naghavi, M. R., Badi, H. N. and Khalighi, A. 2005. Evaluation of Allicin Content and Botanical Traits in Iranian Garlic (Allium sativum L.) Ecotypes. Sci. Hortic., 103:155–166.
8. Balunas, M. J. and Kinghorn, A. D. 2005. Drug Discovery from Medicinal Plants. Life Sci., 78: 431 – 441.
9. Chen, D. X., Cui, G. L., Zhang, X. and Li, L. Y. 2014. Genetic Structure and Genetic Diversity of Artemisia annua Varieties (Strains) Populations Based on SCoT Markers. Zhongguo Zhong Yao Za Zhi., 39: 3254-3258.
10. Davis, S. L. 2001. Phenotypic and Genetic Correlations among Floral Traits in Two Species of Thalictrum. J. Hered., 94: 361–366.
11. Delabays, N., Simonnet, X. and Gaudin, M. 2001. The Genetics of Artemisinin Content in Artemisia annua L. and the Breeding of High Yielding Cultivars. Curr. Med. Chem., 8: 1795-1801.
12. Fathi, A., Ghareyazi, B., Haghnazari, A., Ghaffari, M. R., Pirseyedi, S. M., Kadkhodaei, S., Naghavi, M. R. and Mardi, M. 2008. Assessment of the Genetic Diversity of Almond (Prunus dulcis) Using Microsatellite Markers and Morphological Traits. Iran. J. Biotechnol., 6: 98-106.
13. Ghafoori, H., Sariri, R., Naghavi, M. R., Aryakia, E., Dolatyari, A., Shahzadeh Fazeli, S. A., Ramazani, H. and Farahmand, Z. 2013. Analysis of Artemisinin Isolated from Artemisia annua L. by tlc and hplc. J. Liq. Chromatogr. Relat. Technol., 36: 1198–1206.
14. Govindaraj, M., Vetriventhan, M. and Srinivasan, M. 2015. Importance of Genetic Diversity Assessment in Crop Plants and Its Recent Advances: An Overview of Its Analytical Perspectives. Genet. Res. Int., 1: 1-15.
15. Graham, I., Besser, K., Blumer, S., Branigan, C. A., Czechowski, T., Elias, L., Guterman, I., Harvey, D., Isaac, P. G., Khan, A. M., Larson, T. R., Li, Y., Pawson, T., Penfield, T., Rae, A. M., Rathbone, D. A., Reid, S., Ross, J., Smallwood, M. F., Segura, V., Townsend, T., Vyas, D., Winzer, T. and Bowles, D. 2010. The Genetic Map of Artemisia annua L. Identifies Loci Affecting Yield of the Antimalarial Drug Artemisinin. Sci., 327: 328-331.
16. Haddadi, P., Ebrahimi, A., Langlade, N. B., Yazdi-samadi, B., Berger, M., Calmon, A., Naghavi, M. R., Vincourt, P. and Sarrafi, A. 2012. Genetic Dissection of Tocopherol and Phytosterol in Recombinant Inbred Lines of Sunflower through Quantitative Trait Locus Analysis and the Candidate Gene Approach. Mol. Breed., 29: 717-729.
17. Huang, W. D., Zhao, X. Y., Zhao, X., Zhao, H. L., Wang, S. K. and Lian, J. 2011. A Combined Approach Using ISSR and ITS Analysis for the Characterization of Artemisia halodendron from Horqin Sandy Land, Northern China. Biochem. Sys. Ecol., 39: 346–351.
18. Humphreys, M. O. 2003. Utilization of Plant Genetic Resources in Breeding for Sustainability. Plant Genet. Res. Character. Util., 1: 11–18.
19. Jessing, K. K., O. Duke, S. and Cedergreeen, N. 2014. Potential Ecological Roles of Artemisinin Produced by Artemisia annua L. J. Chem. Ecol., 40: 100–117.
20. Jump, A. S., Marchant, R. and Penuelas, J. 2008. Environmental Change and the Option Value of Genetic Diversity. Trends Plant Sci., 14: 51–58.
21. Keivani, M., Ramezanpour, S. S., Soltanloo, H., Choukan, R., Naghavi, M. R. and Ranjbar, M. 2010. Genetic Diversity Assessment of Alfalfa (Medicago sativa L.) Populations Using AFLP Markers. Aust. J. Crop Sci., 4: 491-497.
22. Kumar, L. S., Jena, S. N. and Nair, K. N. 2010. ISSR Polymorphism in Indian Wild Orange (Citrus indica Tanaka, Rutaceae) and Related Wild Species in North-East India. Sci. Hort., 123: 350–359.
23. Liu, Y., Zhang, J. M., Wang, X. G., Liu, F. and Shen, Z. B. 2013. Genetic Diversity in Vicia amoena (Fabaceae) Germplasm Resource in China Using SRAP and ISSR Markers. Biochem. Syst. Ecol., 51: 86−93.
24. Mannan, A., Ahmed, I., Arshad, W., Asim, M. F., Qureshi, R. A., Hussain, I. Mirza. B. 2010. Survey of Artemisinin Production by Diverse Artemisia Species in Northern Pakistan. Malaria J., 9: 310.
25. Mclellan, T. 2005. Correlated Evolution of Leaf Shape and Trichomes in Begonia dregei (Begoniaceae). Am. J. Bot., 92: 1616-1623.
26. Moghaddam, M., Omidbiagi, R. and Naghavi, M. R. 2011. Evaluation of Genetic Diversity among Iranian Accessions of Ocimum spp. Using AFLP Markers. ‎Biochem. Syst. Ecol., 39: 619-626.
27. Naghavi, M. R., Alaeimoghadam, F. and Ghafoori, H. 2014. Artemisia Species from Iran as Valuable Resources for Medicinal Uses. World Acad. Sci. Eng. Technol., 11: 1058-1064.
28. Naghavi, M. R. and Jahansouz, M. R. 2005. Variation in the Agronomic and Morphological Traits of Iranian Chickpea Accessions. J. Integ. Plant Biol., 47: 375-379.
29. Nasiri, J., Naghavi, M. R., Alizadeh, H., Moghadam, M. R. F. 2016. Seasonal-based Temporal Changes Fluctuate Expression Patterns of TXS, DBAT, BAPT and DBTNBT Genes alongside Production of Associated Taxanes in Taxus baccata. Plant Cell Rep., 35: 1103-1119.
30. Nazar, N. and Mahmood, T. 2011. Morphological and Molecular Characterization of Selected Artemisia Species from Rawalakot, Azad Jammu and Kashmir. Acta Physiol. Plant., 33: 625-633.
31. Ollerton, J and Lack, A. 1998. Relationships between Flowering Phenology, Plant Size and Reproductive Success in Lotus corniculatus (Fabaceae). Plant Ecol., 139: 35-47.
32. Parker, I. M., López, I., Petersen, J. J., Anaya, N., Cubilla-Rios, L. and Potter, D. 2010. Domestication Syndrome in Caimito (Chrysophyllum cainito L.): Fruit and Seed Characteristics. Econ. Bot., 64: 161-175.
33. Pazouki, L., Mardi, M., Shanjani, P. S., Hagidimitriou, M., Pirseyedi, S. M., Naghavi, M. R., Avanzato, D., Vendramin, E., Kafkas, S., Ghareyazie, B., Ghaffari, M. R. and Nekoui, S. M. K. 2010. Genetic Diversity and Relationships among Pistacia Species and Cultivars. ‎Conserv. Genet., 11: 311-318.
34. Podlech, D. 1986. Artemisia. In: “Flora Iranica”, (Ed.): Rechinger, K. H. Graz, Academische Druck Verlagsansalt, PP. 159-223.
35. Ranjbar, M., Naghavi, M. R. and Alizadeh, H. 2014. Comparative Analysis of ADS Gene Promoter in Seven Artemisia Species. ‎J. Genet., 93: 767-774.
36. Ranjbar, M., Naghavi, M. R., Alizadeh, H. and Soltanloo, H. 2015. Expression of Artemisinin Biosynthesis Genes in Eight Artemisia Species at Three Developmental Stages. Ind. Crops Prod., 76: 836-843.
37. Ridder, S., Kooy, F. and Verpoorte, R. 2008. Artemisia annua as a Self-Reliant Treatment for Malaria in Developing Countries. J. Ethnopharmacol., 120: 302-314.
38. Singh. N. D., Kumar, S. and Daniell, H. 2015. Expression of b-Glucosidase Increases Trichome Density and Artemisinin Content in Transgenic Artemisia annua Plants. Plant Biotechnol. J., 14: 1-12.
39. Spagnuolo, V., Muscariello, L., Cozzolino, S., Castaldo Cobianchi, R. and Giordano, S. 2007. Ubiquitous Genetic Diversity in ISSR Markers between and within Populations of the Asexually Producing Moss Pleurochaete squarrosa. Plant Ecol., 188: 91-101.
40. Spring, O. 2000. Chemotaxonomy Based on Metabolites from Glandular Trichomes. Adv. Bot. Res., 31:153-174.
41. Tesfaye, K., Govers, K., Bekele, E. and Borsch, T. 2014. ISSR Fingerprinting of Coffea arabica throughout Ethiopia Reveals High Variability in Wild Populations and Distinguishes Them from Landraces. Plant Syst. Evol., 300: 881-897.
42. Thul, S. T., Darokar, M. P., Shasany, A. K. and Khanuja, S. P. 2012. Molecular Profiling for Genetic Variability in Capsicum Species Based on ISSR and RAPD Markers. Mol. Biotechnol., 51: 137-147.
43. Valverde, P. L., Fornoni, J. and Nunez-Farfan, J. 2001. Defensive Role of Leaf Trichomes in Resistance to Herbivorous Insects in Datura stramonium. J. Evol. Biol., 14: 424-432.
44. Wang, X., Zhao, F., Hu, Z., T. Critchley, A., L. Morrell, S. and Duan, D. 2008. Inter-Simple Sequence Repeat (ISSR) Analysis of Genetic Variation of Chondrus crispus Populations from North Atlantic. Aquat. Bot., 88: 154-159.
45. Yang, M., Liu, D., Shao, A., Jin, H., Wang, X., Fan, Z. and Huang, L. 2010. Diversity and Cluster Analysis on Agronomic Traits of Artemisia annua Germplasm Resources in Yun-Gui Plateau. Zhongguo Zhong Yao Za Zhi., 35: 3097-3102.
Journal of Agricultural Science and Technology
Volume 21, Issue 5
September and October 2019
Pages 1265-1276