In vitro Culture of Endemic Astragalus gymnolobus Fischer and Comparison of its Antibacterial, Antioxidant, and Phenolic Profiles with Field Grown Plants

Document Type : Original Research

Authors
A. Yildirim 1
E. Uyar 2
A. Turker 2
1 Department of Field Crops, Faculty of Agricultural and Environmental Science, Bolu Abant Izzet Baysal University, Bolu, Turkey.
2 Department of Biology, Faculty of Science and Art, Bolu Abant Izzet Baysal University, Bolu, Turkey.
Abstract
Astragalus gymnolobus Fischer (Leguminosae family) is an endemic plant to Turkey. Firstly, an in vitro regeneration system was achieved using leaf and petiole explants on Murashige and Skoog (MS) medium supplemented with different cytokinins [Thidiazuron (TDZ), Kinetin (KIN), Benzyladenine (BA)], auxins [Indole-3-Butyric Acid (IBA), Indole-3-Acetic Acid (IAA), 2,4-Dichlorophenoxyacetic acid (2,4-D) and Naphthalene Acetic Acid (NAA)] and Gibberellic Acid (GA3) at diverse concentrations. Best shoot formation was obtained with leaf explants and only TDZ alone or TDZ and IAA combinations were effective for shoot forming. The best shoot inducing response (17.60 shoots per explant at 23.81% shoot frequency) was recorded at 0.5 mg L-1 TDZ alone. Among the TDZ stand-alone treatments, 0.05 and 0.1 mg L-1 TDZ were also effective in terms of shoot induction frequency (31.82 and 30.43%, respectively). Root formation was obtained after 3 months (3.5 roots per shoot with 77.8% root frequency) only in auxin free MS medium (control) when regenerated shoots were cut off 0.5-1 cm from the base to exclude the callus part. This study also aimed to determine and compare the antibacterial and antioxidant properties and phenolic composition of in vitro-regenerated and field-grown A. gymnolobus. The results revealed that field-grown leaves generally showed higher biological activities and had higher amounts of phenolic compounds. Furthermore, A. gymnolobus leaves were noteworthy sources of rutin. This initial in vitro culture protocol for endemic A. gymnolobus is valuable for genetic resources conservation and can be used in stress application studies to increase the level of phenolic substances in in vitro-grown plants.

Keywords

Subjects

1. Adigüzel, A., Sökmen, M., Özkan, H, Ağar, G., Güllüce, M. and Şahin, F. 2009. In vitro Antimicrobial and Antioxidant Activities of Methanol and Hexane Extract of Astragalus Species Growing in the Eastern Anatolia Region of Turkey. Turk. J. Biol., 33: 65–71.
2. Akan, H. and Civelek, Ş. 2001. Astragalus aytatchii (Fabaceae), A New Species from Anatolia, Turkey. Ann. Bot. Fennici., 38: 167–170.
3. Albayrak, S. and Kaya, O. 2018. Antioxidant and Antimicrobial Activities of Four Astragalus species growing Wild in Turkey. Turk. J. Biochem., 43: 425-434.
4. Andrews, J.M. 2009. BSAC Standardized Disc Susceptibility Testing Method (Version 8). J. Antimicrob. Chemother., 64: 454–489.
5. Basalma, D., Uranbey, S., Gürlek, D. and Özcan, S. 2008. TDZ-Induced Plant Regeneration in Astragalus cicer L. Afr. J. Biotechnol., 7: 955–959.
6. Bedir, E., Calis, I., Dunbar, C., Sharan, R., Buolamwini, J.K. and Khan, I.A. 2001. Two Novel Cycloartane-Type Triterpene Glycosides from the Roots of Astragalus prusianus. Tetrahedron, 57: 5961–5966.
7. Bisignano, G., Iauk, L., Kirjavainen, S and Galati, E.M. 1994. Anti-inflammatory, Analgesic, Antipyretic and Antibacterial Activity of Astragalus siculus Biv. Int. J. Pharmacogn., 32: 400–405.
8. Blois, M.S. 1958. Antioxidant Determinations by the Use of a Stable Free Radical. Nature, 181: 1199.
9. Bourezzane, S., Haba, H., Long, C. and Benkhaled, M. 2018. Chemical Composition and Antioxidant Activity of Astragalus monspessulanus L. Growing in Semiarid Areas of Algeria. J. Serb Chem. Soc., 83: 31–38.
10. Chang, C.C., Yang, M.H., Wen, H.M. and Chern, J.C. 2002. Estimation of Total Flavonoid Content in Propolis by Two Complementary Colorimetric Methods. J. Food Drug Anal., 10: 178–182.
11. Davis, P. 1970. Flora of Turkey and the East Aegean Islands. Vol 3. Edinburgh University Press, England, 248 PP.
12. Dilaver, Z., Mirzapour, M. and Kendir, H. 2017. Breaking Seed Dormancy and Micropropagation of Perennial Vulneraria Milkvetch (Astragalus vulnerariae DC.). Acta Sci. Pol. Cultus., 16: 79–88.
13. Erisen, S., Atalay, E. and Yorgancilar, M. 2011. The Effect of Thidiazuron on the in vitro Shoot Development of Endemic Astragalus cariensis in Turkey. Turk. J. Bot., 35: 521–526.
14. Erisen, S., Yorgancilar, M., Atalay, E., Babaoglu, M. and Duran, A. 2010. Callus Induction and Plant Regeneration of the Endemic Astragalus nezaketae in Turkey. Electron. J. Biotechnol., 13: 13–14.
15. Esmaeili, G., Azizi, M., Aroei, H., and Samiei, L. 2016. Micropropagation of Astragalus adscendens: A Source of Gaz-angabin Manna in Iran (Persian manna). J. Agric. Sci .Technol., 18: 741–750.
16. Hasancebi, S., Kara, N.T., Çakır, Ö. and Arı, Ş. 2011. Micropropagation and Root Culture of Turkish Endemic Astragalus chrysochlorus (Leguminosae). Turk. J. Bot., 35: 203–210.
17. Haşimi, N., Ertaş, A., Yılmaz, M.A., Boğa, M., Temel, H., Demirci, S., Özden, T.Y., Yener, İ. and Kolak, U. 2017. LC-MS/MS and GC-MS Analyses of Three Endemic Astragalus Species from Anatolia Towards Their Total Phenolic-Flavonoid Contents and Biological Activities. Biol. Divers. Conserv., 10: 18–30.
18. Huetteman, C.A. and Preece, J.E. 1993. Thidiazuron: a Potent Cytokinin for Woody Plant Tissue Culture. Plant Cell Tiss. Org. Cult., 33: 105–119.
19. IUCN. 2015. The IUCN Red List of Threatened Species, Version 2015-4.
20. Jaradat, N.A., Zaid, A.N., Abuzant, A., Khalaf, S. and Abu-Hassan, N. 2017. Phytochemical and Biological Properties of Four Astragalus Species Commonly Used in Traditional Palestinian Medicine. Eur. J. Integr. Med., 9: 1–8.
21. Kanaan, H., El-Mestrah, M., Sweidan, A., As-Sadi, F., Al Bazzal, A. and Chokr, A. 2017. Screening for Antibacterial and Antibiofilm Activities in Astragalus angulosus. J. Intercult. Ethnopharmacol., 6: 50–57.
22. Langari, S. and Salehi, E.A. 2015. Evaluation of the Antioxidant Capacity of Different Extracts of Astragalus glaucacanthus. J. Appl. Environ. Biol. Sci., 4: 14–17.
23. Lock, J.M. and Simpson, K. 1991. Legumes of West Asia: a Check-list. Royal Botanic Gardens, Kew
24. Luo, J.P. and Jia, J.F. 1998. Callus Induction and Plant Regeneration from Hypocotyl Explants of the Forage Legume Astragalus adsurgens. Plant Cell. Rep., 17: 567–570.
25. Murashige, T. and Skoog, F. 1962. A Revised Medium for Rapid Growth and Bio assays with Tobacco Tissue Cultures. Physiol. Plant., 15: 473–497.
26. Pistelli, L., Bertoli, A., Lepori, E., Morelli, I. and Panizzi, L. 2002. Antimicrobial and Antifungal Activity of Crude Extracts and Isolated Saponins from Astragalus verrucosus. Fitoterapia, 73: 336–339.
27. Pistelli, L., Giachi, I., Lepori, E. and Bertoli, A. 2003. Further Saponins and Flavonoids from Astragalus verrucosus Moris. Pharm. Biol., 41: 568–572.
28. Slinkard, K. and Singleton, V.L. 1977. Total Phenol Analysis: Automation and Comparison with Manual Methods. Am. J. Enol. Viticult., 28: 49–55.
29. Teyeb, H., Zanina, N., Neffati, M., Douki, W. and Najjar, M.F. 2012. Cytotoxic and Antibacterial Activities of Leaf Extracts of Astragalus gombiformis Pomel (Fabaceae) Growing Wild in Tunisia. Turk. J. Biol., 36: 53–58.
30. Turgut-Kara, N. and Arı, S. 2006. Micropropagation of Astragalus maximus Willd. Biotechnol. Biotechnol. Equip., 20: 20–22.
31. Turker, A.U. and Koyluoglu, H. 2012. Biological Activities of Some Endemic Plants in Turkey. Rom. Biotechnol. Lett., 17: 6949-6961.
32. Turker, A.U. and Yildirim, A.B. 2013. Evaluation of Antibacterial and Antitumor Activities of Some Turkish Endemic Plants. Trop. J. Pharm. Res., 12: 1003-1010.
33. Turker, H., Yıldırım, A.B., Karakaş, F.P. and Köylüoğlu, H. 2009. Antibacterial Activities of Extracts from Some Turkish Endemic Plants on Common Fish Pathogens. Turk. J. Biol., 33: 73–78.
34. Uranbey, S., Çöçü, S., Sancak, C., Parmaksız, İ., Khawar, K.M., Mirici, S., and Özcan, S. 2003. Efficient Adventitious Shoot Regeneration in Cicer Milkvetch. Biotechnol. Biotechnol. Equip., 17: 33–37.
35. Yorgancilar, M. and Erisen, S. 2011. The Effect of Thidiazuron (TDZ) on Shoot Regeneration of Astragalus schizopterus. J. Anim. Plant Sci., 21: 519–524.
Journal of Agricultural Science and Technology
Volume 22, Issue 3
March and April 2020
Pages 815-828