Beneficial effects of endophytic fungi inoculation on tanshinones and phenolic compounds of Salvia abrotanoides

Document Type : Original Article


1 Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Salvia abrotanoides is considered as a new source of tanshinone-producing plants in Iran. Symbiosis of endophytic fungi with their host plants is an effective tool to promote the growth and secondary metabolism of medicinal herbs. Therefore, using endophytic fungi as a biotic elicitor is a proper solution to increase the yield of plant products. 
Materials and Methods: In this study, some endophytic fungi were first isolated from the root of S. abrotanoides, then two of them (Penicillium canescens and Talaromyces sp.) were co-cultivated with the sterile seedling of S. abrotanoides in pot culture. After proving the colonization of these fungi in the root tissues by microscopic studies, their effects on the production of critical medicinal compounds such as tanshinones and phenolic acids were investigated in the vegetation stage (120 days). 
Results: Our results showed that the content of cryptotanshinone (Cry) and tanshinone IIA (T-IIA) in plants inoculated with P.  canescens increased by 77.00% and 19.64%, respectively, compared with non-inoculated plants (control). The contents of mentioned compounds in plants inoculated with Talaromyces sp. increased by 50.00% and 23.00%, respectively. In this case, in plants inoculated with P. canescens, it was found that the level of caffeic acid, rosmarinic acid, and its PAL enzyme activity increased by 64.00%, 69.00%, and 50.00%, respectively, compared with the control.
Conclusion: Endophytic fungi have specific modes of action and the ability to provide multiple benefits. Each of the two strains is a highly considerable microbial resource for the growth and accumulation of active compounds of S. abrotanoides.


1. Ye B, Wu Y, Zhai X, Zhang R, Wu J, Zhang C, et al. Beneficial effects of endophytic fungi from the Anoectochilus and Ludisia species on the growth and secondary metabolism of Anoectochilus roxburghii. ACS omega 2020;5:3487-3497.
2. Venieraki A, Dimou M, Katinakis P. Endophytic fungi residing in medicinal plants have the ability to produce the same or similar pharmacologically active secondary metabolites as their hosts. HPPJ 2017;10:51-66.
3. Satheesan J, Narayanan AK, Sakunthala M. Induction of root colonization by Piriformospora indica leads to enhanced asiaticoside production in Centella asiatica. Mycorrhiza 2012; 22:195-202.
4. Zhang FS, Lv YL, Zhao Y, Guo SX. Promoting role of an endophyte on the growth and contents of kinsenosides and flavonoids of Anoectochilus formosanus Hayata, a rare and threatened medicinal Orchidaceae plant. J Zhejiang Univ Sci B 2013;14:785-792.
5. Ghahreman A. 1975-2000. Colored Flora of Iran, Vol. 1-20.
6. Ma P, Liu J, Osbourn A, Dong J, Liang Z. Regulation and metabolic engineering of tanshinone biosynthesis. RSC Adv 2015;5:18137-18144.
7. Sairafianpour M, Christensen J, Stærk D, Budnik BA, Kharazmi A, Bagherzadeh K, Jaroszewski JW. Leishmanicidal, antiplasmodial, and cytotoxic activity of novel diterpenoid 1, 2 -quinones from Perovskia abrotanoides: new source of tanshinones. J Nat Prod 2001 ;64:1398-1403. 
8. Zaker A, Sykora C, Gössnitzer F, Abrishamchi P, Asili J, Mousavi SH, et al. Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel. Ind Crops Prod 2015; 67:97-102.
9. Mahboubi M. Iranian medicinal plants as antimicrobial agents. J Microbiol Biotechnol Food Sci 2013;2:2388-2405.
10. Kumar G P, Gupta S, Murugan M P, Bala Singh S. Ethnobotanical studies of Nubra Valley-A cold arid zone of Himalaya. Ethnobot leafl 2009;2009:9.
11. Tareen RB, Bibi T, Khan MA, Ahmad M, Zafar M, Hina S. Indigenous knowledge of folk medicine by the women of Kalat and Khuzdar regions of Balochistan, Pakistan. Pak J Bot 2010;42:1465-1485.
12. Zaker A, Asili J, Abrishamchi P, Tayarani-Najaran Z, Mousavi SH. Cytotoxic and apoptotic effects of root extract and tanshinones isolated from Perovskia abrotanoides Kar. Iran J Basic Med Sci 2017;20:1377-1384.
13. Jaafari MR, Hooshmand S, Samiei A, Hossainzadeh H. Evaluation of-leishmanicidal effect of Perovskia abrotanoides Karel root extract by in vitro leishmanicidal assay using promastigotes of Leishmania major. Pharmacol Online 2007;1:299-303.
14. Mahanty T, Bhattacharjee S, Goswami M, Bhattacharyya P, Das B, Ghosh A, et al. Biofertilizers: A potential approach for sustainable agriculture development. Environ Sci Pollut Res Int 2017;24:3315-3335.
15. Fisher P, Petrini O, Sutton B. A comparative study of fungal endophytes in leaves, xylem and bark of Eucalyptus in Australia and England. Sydowia 1993;45:338-345.
16. Teimoori-Boghsani Y, Ganjeali A, Cernava T, Müller H, Asili J, Berg G. Endophytic fungi of native Salvia abrotanoides plants reveal high taxonomic diversity and unique profiles of secondary metabolites. Front Microbiol 2020; 10:3013.
17. Babu AG, Kim SW, Yadav DR, Hyum U, Adhikari M, Lee YS. Penicillium menonorum: A novel fungus to promote growth and nutrient management in cucumber plants. Mycobiology 2015;43:49-56.
18. Michal Johnson J, Sherameti I, Ludwig A, Nongbri PL, Sun C, Lou B, et al. Protocols for Arabidopsis thaliana and Piriformospora indica co-cultivation-A model system to study plant beneficial traits. Endocytobiosis Cell Res. ISE. 2011:101-113.
19. Trouvelot A, Kough J, Gianinazzi-Pearson V, editors. Mesure du taux de mycorhization VA d’un système radiculaire. Recherche de méthode d’estimation ayant une signification fonctionnelle. Physiological and genetical aspects of mycorrhizae: Proceedings of the 1st european symposium on mycorrhizae. 1985; 217-221.
20. Wang S, Zhang S, Zhou T, Zeng J, Zhan J. Design and application of an in vivo reporter assay for phenylalanine ammonia-lyase. Appl Microbiol Biotechnol 2013;97:7877-7885.
21. Juliani HR, Koroch AR, Simon JE. Basil: A source of rosmarinic acid. ACS Publications; 2008.
22. Srivastava S, Somasagara RR, Hegde M, Nishana M, Tadi SK, Srivastava M, et al. Quercetin, a natural flavonoid interacts with DNA, arrests cell cycle and causes tumor regression by activating mitochondrial pathway of apoptosis. Sci Rep 2016;6:1-13.
23. Compant S, Clément C, Sessitsch A. Plant growth-promoting bacteria in the rhizo-and endosphere of plants: Their role, colonization, mechanisms involved and prospects for utilization. Soil Biol Biochem 2010;42:669-678.
24. Yan L, Zhu J, Zhao X, Shi J, Jiang C, Shao D. Beneficial effects of endophytic fungi colonization on plants. Appl Microbiol Biotechnol 2019;103:3327-3340.
25. Qayyum MA, Wakil W, Arif MJ, Sahi ST, Dunlap CA. Infection of Helicoverpa armigera by endophytic Beauveria bassiana colonizing tomato plants. Biol Control 2015;90: 200-207.
26. Di P, Zhang L, Chen J, Tan H, Xiao Y, Dong X, et al. 13C tracer reveals phenolic acids biosynthesis in hairy root cultures of Salvia miltiorrhiza. ACS Chem Biol 2013;8:1537-1548.
27. Gao W, Sun H-X, Xiao H, Cui G, Hillwig ML, Jackson A, et al. Combining metabolomics and transcriptomics to characterize tanshinone biosynthesis in Salvia miltiorrhiza. BMC Genomics 2014;15:1-14.
28. Song J, Wang Z. Molecular cloning, expression and characterization of a phenylalanine ammonia-lyase gene (SmPAL1) from Salvia miltiorrhiza. Mol Biol Rep 2009;36:939-952.
29. Yu H-N, Liu X-Y, Gao S, Han X-J, Cheng A-X, Lou H-X. Molecular cloning and functional characterization of a phenylalanine ammonia-lyase from liverwort Plagiochasma appendiculatum. Plant Cell Tissue Organ Cult 2014;117:265-277.
30. Zhang S, Yan Y, Wang B, Liang Z, Liu Y, Liu F, et al. Selective responses of enzymes in the two parallel pathways of rosmarinic acid biosynthetic pathway to elicitors in Salvia miltiorrhiza hairy root cultures. J Biosci Bioeng 2014;117:645-651.
31. Yang D, Huang Z, Xing B, Jin W, Yan X, Guo Z, et al. Regulation of folic acid on phenolic acids production in Salvia miltiorrhiza hairy roots. Plant Cell Tissue Organ Cult 2016;127:175-185.
32. Zhou LS, Tang K, Guo SX. The plant growth-promoting fungus (PGPF) Alternaria sp. A13 markedly enhances Salvia miltiorrhiza root growth and active ingredient accumulation under greenhouse and field conditions. Int J Mol Sci 2018;19:270.
33. Zhai X, Luo D, Li X, Han T, Jia M, Kong Z, et al. Endophyte Chaetomium globosum D38 promotes bioactive constituents accumulation and root production in Salvia miltiorrhiza. Front Microbiol 2018; 8:2694.
34. Chen HM, Wu HX, He XY, Zhang HH, Miao F, Liang ZS. Promoting tanshinone synthesis of Salvia miltiorrhiza root by a seed endophytic fungus, Phoma herbarum D603. Zhongguo Zhongyao Zazhi 2020;45:65-71.