بررسی اثر قارچ میکوریزا (Piriformospora indica) بر صفات مورفولوژیکی، فیزیولوژیکی و بیوشیمیایی گیاه دارویی استویا (Stevia rebaudiana) تحت تنش خشکی

نوع مقاله : مقالات پژوهشی

نویسندگان

• اسمعیل نبی زاده ¹
• مسعود حق شناس ²
• خدیجه احمدی ³

¹ دانشگاه آزاد اسلامی مهاباد، مهاباد، ایران

² دانشگاه محقق اردبیلی، اردبیل، ایران

³ دانشگاه شاهد، تهران، ایران

https://doi.org/10.22067/jhs.2022.75337.1173

چکیده

گیاه دارویی استویا از خانواده Asteraceae، حاوی ترکیبات طبیعی، به ویژه استویوزید و ریبائودیوزید A است که تخمین زده می­شود 150 تا 400 برابر شیرین­تر از ساکاروز باشد. به­منظور بررسی اثر قارچ اندوفیت Piriformospora indica در شرایط تنش آبی بر خصوصیات رویشی، پارامترهای فیزیولوژیکی و عناصر ریزمغذی گیاه دارویی استویا، آزمایشی به­صورت فاکتوریل در قالب طرح کاملاً تصادفی در چهار تکرار در سال 97-1396 اجرا شد. عامل اول تنش خشکی در چهار سطح (25، 45، 60 و 80 درصد ظرفیت مزرعه) و عامل دوم تلقیح نشاء با قارچ در دو سطح (عدم تلقیح و تلقیح باP. indica) بودند. صفات مورد مطالعه شامل کولونیزاسیون ریشه، وزن خشک، تعداد برگ، ارتفاع بوته، قطر ساقه، کلروفیل a، b، کلروفیل کل، کاروتنوئید، پرولین، قندهای محلول، قدرت آنتی­اکسیدانی و عناصر ریزمغذی شامل مس، آهن، روی و منگنز بودند. نتایج نشان داد که صفات مورد ارزیابی در پژوهش حاضر تحت تأثیر تیمارهای اصلی قارچ و تنش خشکی قرار گرفتند. نهال­هایی که با قارچ اندوفیت P. indica تلقیح شده دارای بیش­ترین درصد کولونیزاسیون ریشه، پارامترهای رشد، محتوای رنگیزه­های فتوسنتزی، ترکیبات محلول و عناصر ریزمغذی نسبت به عدم تلقیح بودند. تنش خشکی موجب افزایش قندهای محلول، میزان پرولین و قدرت آنتی­اکسیدانی برگ گیاه دارویی استویا شد و با افزایش تنش خشکی از 80 درصد ظرفیت زراعی به 25 درصد باعث کاهش مابقی صفات شد. طبق نتایج مقایسه میانگین اثر برهمکنش قارچ در تنش خشکی، بیش­ترین میزان کولونیزاسیون ریشه، قطر ساقه  و محتوای کاروتنوئید در تلقیح گیاه با قارچ P. indica و تنش 25 درصد و قدرت آنتی­اکسیدانی در تنش 80 درصد مشاهده شد. باتوجه به نتایج تحقیق حاضر استفاده از قارچ P. indica بیش­ترین تأثیر مثبت بر خصوصیات کمی و کیفی گیاه دارویی استویا نسبت به عدم تلقیح قارچ برخوردار بود.

کلیدواژه‌ها

• آنتی‌اکسیدان
• استویا
• عناصر غذایی
• کم آبی
• کلونیزاسیون

20.1001.1.20084730.1402.37.2.12.8

موضوعات

• گیاهان دارویی

عنوان مقاله [English]

The Effect of Mycorrhizal Fungus (Piriformospora indica) on the Morphological, Physiological and Biochemical Traits of the Medicinal Plant Stevia (Stevia rebaudiana) under Drought Stress

نویسندگان [English]

• Esmaeil Nabizadeh ¹
• Masud Haghshenas ²
• Khadijeh Ahmadi ³

¹ Islamic Azad University, Mahabad, Mahabad, Iran

² Mohaghegh Ardabili University, Ardabil, Iran

³ Shahed University, Tehran, Iran

چکیده [English]

Introduction
Stevia (Stevia rebaudiana) is one of the medicinal plants of the Asteraceae family that contains natural compounds, especially stevioside and ribaodioside A, which are estimated to be 150 to 400 times sweeter than sucrose. Plants are exposed to various environmental stresses during growth and development under natural and agricultural conditions. Among these, drought is one the most severe environmental stresses affecting plant productivity. About 80–95% of the fresh biomass of the plant body is comprised of water, which plays a vital role in various physiological processes including many aspects of plant growth, development, and metabolism. Stevia is susceptible to various environmental stresses but the major effects are contributed by drought. Today, the fungal species Stevia rebaudiana is used as a biofertilizer and increases the production of secondary metabolites of economically valuable plants and also increases the growth and seed production of many plants. This fungal endophyte produces a significant amount of acid phosphatase for mobility in a wide range of insoluble or complex forms of phosphate, enabling the host plant to have adequate access to inactive phosphorus reserves in the soil. However, medicinal plants that are cultivated have often been reported to have lower abundance of arbuscular mycorrhizal fungi in the rhizosphere, which significantly reduces plant survival. Considering the coexistence role of mycorrhizal fungi in modulating the effects of drought stress, the aim of this study was to investigate the morphological, physiological and biochemical traits of stevia in response to the effects of mycorrhizal inoculation and drought stress.
 
Materials and Methods
This experiment was conducted to investigate the effect of P. indica endophytic fungus under water stress conditions on vegetative characteristics, physiological parameters and micronutrients of stevia. A factorial experiment was employed based a completely randomized design with four replications in the research greenhouse of Islamic Azad University, Mahabad Branch in 2017. The first factor was drought stress at four levels (25, 45, 60 and 80% of field capacity) and the second factor was inoculation of seedlings with fungus at two levels (no inoculation and inoculation with P. indica). Water stress was applied based on a combination of plant appearance symptoms (no wilting to severe wilting) and soil moisture. Investigated traits included root colonization, dry weight, leaf number, plant height, stem diameter, chlorophyll a, b, total chlorophyll, carotenoids, proline, soluble sugars, antioxidant power and micronutrients including copper, iron, zinc and manganese. To analyze the data variance, SAS 9.1 statistical software was used to analyze the variance of the data.
Results and Discussion
The results showed that the evaluated traits in the present study were affected by the main treatments of fungus and drought stress. Seedlings inoculated with P. indica endophytic fungi had the highest percentage of root colonization, growth parameters, photosynthetic pigment content, soluble compounds and micronutrients compared to no inoculation. Drought stress increased soluble sugars, proline content and antioxidant power of stevia leaves and decreased the other traits by increasing the stress level from 25 to 80%. The highest rate of root colonization (26.90%), stem diameter (3.21 mm) and carotenoid content (1.71 μg/ml) was observed in the treatment of plant inoculation with fungi and 25% drought stress. While the highest antioxidant power was found in the treatment of plant inoculation with fungi and 80% drought stress. According to the results of the present study, use of P. indica fungus had the most positive effect on the quantitative and qualitative characteristics of stevia medicinal plant compared to no fungus inoculation.
 
Conclusion
This study showed the positive effect of P. indica endophyte inoculation on quantitative and qualitative characteristics of root colonization, dry weight, number of leaves, plant height, stem diameter, chlorophyll a, b, total chlorophyll, carotenoids, proline, soluble sugars, antioxidant power and The micronutrients of calcium, iron, zinc and manganese showed stevia, and drought stress reduced the studied traits except for proline content, soluble sugars and antioxidant power. Inoculation of stevia seedlings with P. indica endophytic fungi at drought stress levels had the highest rate of root colonization, stem diameter, carotenoid content and antioxidant power compared to non-fungal inoculation. Therefore, due to the effect of biological compounds of natural origin and the production of plants with healthier active secondary compounds, the use of P. indica endophytic fungi can be recommended.

کلیدواژه‌ها [English]

• Nutrients
• Pigment
• Photosystem
• Stevia
• Water deficit

مراجع

1. Abdollahi, , Ali Asgharzad, N., Zahtab Selmasi, S., & Khoshru, B. (2019). Effects of endophytic fungus piriformospora indica on growth indices and nutrient uptake by anise plant (Pimpinella anisum) under water deficit stress conditions. Journal of Agricultural Science and Sustainable Production, 29(4), 51-64.
2. Arabzadeh, N. (2012). The effect of drought stress on soluble carbohydrates (sugars) in two species of Haloxylon persicum and Haloxylon aphyllum. Asian Journal of Plant Sciences, 11, 44-51. https://doi.org/10.3923/ajps.2012.44.51
3. Baligar, V.C., Fageria, N.K., & He, Z.L. (2001). Nutrient use efficiency in plants. Communications in Soil Science and Plant Analysis, 32, 921-950.
4. Bates, S., Waldern, R.P., & Teave, I.D. (1973). Rapid determination of free proline for water stress studies. Plant and Soil, 39, 205-207. https://doi.org/10.1007/BF00018060
5. Benzie, F.F., & Strain, J.J. (1996). ̍The Ferric reducing ability of plasma (FRAP) as a measure of ‘antioxidant power: the FRAP assay. Analytical Biochemistry, 239, 70–76. https://doi.org/10.1006/abio.1996.0292
6. Bhuyan, K., Bandyopadhyay, P., Kumar, P., Mishra, D.K., Prasad, R., Kumari, A., & Yadava, P.K. (2015). Interaction of Piriformospora indica with Azotobacter chroococcum. Scientific Reports, 5, 13911. https://doi.org/10.1038/srep13911
7. Brodersen, R., Roddy, A.B., Wason, J.W., & McElrone, A.J. (2019). Functional status of xylem through time. Annual Review Plant Biology, 70, 407–433. https://doi.org/10.1146/annurev-arplant-050718-100455
8. Cheng, Q., Zou, Y.N., Wu, Q.S., & Kuča, K. (2021). Arbuscular mycorrhizal fungi alleviate drought stress in trifoliate orange by regulating H+-ATPase activity and gene expression. Frontiers in Plant Science, 12, 659-694. https://doi.org/10.1016/10.3389/fpls.2021.659694
9. Costache, A., Campeanu, G., & Neata, G. (2012). Studies concerning the extraction of chlorophyll and total carotenoids from vegetables. Romanian Biotechnological Letters, 17(5), 7702-7708.
10. Devincentis, J. (2020). Scales of sustainable agricultural water management. Ph.D. Thesis, University of California, Davis, CA, USA.
11. Diatta, A., Fike, J.H., Battaglia, M.L., Galbraith, J., & Baig, M.B. (2020). Effects of biochar on soil fertility and crop productivity in arid regions: A review. Arabian Journal of Geosciences, 13, 595. https://doi.org/10.1007/s12517-020-05586-2
12. DuBois, , Gilles, K.A., Hamilton, J.K., Rebers, P.A., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. American Chemical Society, 28(3), 350–356
13. Eziz, A, Yan, Z, Tian, D, Han, W, Tang, Z., & Fang, J. (2017). Drought effect on plant biomass allocation: A meta-analysis. Ecology and Evolution, 7, 11002–11010. https://doi.org/10.1002/ece3.3630
14. Gao, X., & He, X.L. (2007). Ecological study on am fungi around roots of medicinal plants in the middle area of Hebei province. Agricultural Research in the Arid Areas, 25(3), 196-202. https://doi.org/10.1016/S1872-2040(07)60079-6
15. Giovannetti, M., & Mosse, B. (1980). An evaluation of techniques for measuring vesicular Arbuscular Mycorrhizal infection in roots. New Philologists, 84, 489-500. https://doi.org/10.1111/j.1469-8137.1980.tb04556.x
16. Gosal, S., Wani, S.H., & Kang, M.S. (2010). Biotechnology and crop improvement. Journal of Crop Improvement, 24(2), 153-217. https://doi.org/10.1080/15427520903584555
17. Haghir Ebrahimabadi, A., Hatami, M., Karimzadeh Asl, K., & Ghorbanpour, M. (2018). Effect of mycorrhizal fungi and biophosphor fertilizer on growth features, yield and yield components, and essntial oil constituents in Cuminum cyminum Journal of Medicinal Plants, 17(66), 74-90. (In Persian with English abstract). https://doi.org/20.1001.1.2717204.2018.17.66.3.1
18. Hajihashemi, , & Ehsanpour, A.A. (2014). Antioxidant response of Stevia rebaudiana B. to polyethylene glycol and paclobutrazol treatments under in vitro culture. Applied Biochemistry and Biotechnology, 172(8), 4038-4052. (In Persian with English abstract)
19. Hamzei, , & Salimi, F. (2015). Root colonization, yield and yield components of milk thistle (Silybum marianum) affected by mycorhizal fungi and phosphorus fertilizer. Journal of Agricultural Science and Sustainable Production, 24(4), 85-96.
20. Hill, W., & Kafer, E. (2001). Improved protocols for Aspergillus minimal medium: trace element and minimal medium salt stock solutions. Fungal Genetics Reports, 48, 8. https://doi.org/10.4148/1941-4765.1173
21. Lee, H., Kim, Y.S., & Lee, C.B. (2001). The inductive responses of the antioxidant enzymes by salt stress in rice (Oryza sativa L.). Jurnal Plant Physioloy, 158, 737–745. https://doi.org/10.1078/0176-1617-00174
22. Lemus-Mondaca, R., Vega-Gálvez, A., Zura-Bravo, L., & Ah-Hen, K. (2012). Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehensive review on the biochemical, Nutritional and Functional Aspects. Food Chemistry, 132(3), 1121-1132. https://doi.org/10.1016/j.foodchem.2011.11.140
23. Ma, F., Yang, X.H., Li, P.M., & Tong, R.J. (2005). Investigation of the diversity of arbuscular mycorrhizal structure of medicinal plants in Chongqing. Journal of Southwest Agricultural University, 27(3), 406-409.
24. Meng, L., He, J.D., Zou, Y.N., Wu, Q.S., & Kuča, K. (2020). Mycorrhiza-released glomalin-related soil protein fractions contribute to soil total nitrogen in trifoliate orange. Plant, Soil and Environment, 66, 183-189. https://doi.org/10.17221/100/2020-PSE
25. OConnell, (2017). Towards adaptation of water resource systems to climatic and socio-economic Chang. Water Resources Management, 31, 2965–2984. https://doi.org/10.1007/s11269-017-1734-2
26. Okorie, O., Mphambukeli, T.N., & Amusan, S.O. (2019). Exploring the political economy of water and food security nexus in BRICS. Africa Insight, 48, 21–38. https://hdl.handle.net/10520/EJC-15b208a68b
27. Ortas, , Sari, N., Akpinar, C., & Yetisir, H. (2011). Screening mycorrhiza species for plant growth, P and Zn uptake in pepper seedling grown under greenhouse conditions. Scientia Horticulturae, 128, 92–98. https://doi.org/10.1016/j.scienta.2010.12.014
28. Phillips, M., & Hayman, D.S. (1970). Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British Mycological Society, 55(1), 158-161. https://doi.org/10.1016/S0007-1536(70)80110-3
29. Rathinasabapathi, (2000). Metabolic engineering for stress tolerance: Installing osmoprotectant synthesis pathways. Annals of Botany, 86, 709-716. https://doi.org/10.1006/anbo.2000.1254
30. Sanayei, , Barmaki, M., Ebadi khazine Gadim, A., & Torabi Giglou, M. (2021). Effect of drought stress and inoculation of mycorrihizal fungi and Pseudomonas spp. on some morpho-physiological characteristics of roselle (Hibiscus sabdaeiffa L.). Journal of Agricultural Science and Sustainable Production, 30(2), 71-89.
31. Sanmartín, , Garmendia, I., Romano, B., Díaz, M., Palop, J.A., & Goicoechea, N. (2014). Mycorrhizal inoculation affected growth, mineral composition, proteins and sugars in lettuces biofortified with organic or inorganic selenocompounds. Scientia Horticulturae, 180, 40–51. https://doi.org/10.1016/j.scienta.2014.09.049
32. Seyedmohammadi, , Barmaki, M., & Davari, M. (2019). Effect of mycorrhizal fungi on leaf yield, root colonization percentage and some features of Stevia rebaudiana root in a soilless culture system. Journal of Agricultural Science and Sustainable Production, 29(2), 189-204.
33. Sherameti, , Tripathi, S., Varma, A., & Oelmüller, R. (2008). The root-colonizing endophyte Pirifomospora indica confers drought tolerance in Arabidopsis by stimulating the expression of drought stress–related genes in leaves. Molecular Plant-Microbe Interactions, 21(6), 799-807. https://doi.org/10.1094/MPMI-21-6-0799
34. Singh,, Sharma, J., Rexer, K.H., & Varma, A. (2000). Plant productivity determinants beyondminerals, water and light: Piriformospora indica – A revolutionary plant growth promoting fungus. Current Science, 79(11), 1548-1554.
35. Sirrenberg, , Göbel, C., Grond, S., Czempinski, N., Ratzinger, A., Petr Karlovsky, P., Patricia Santos, P., Feussner, L., & Pawlowski, K. (2007). Piriformospora indicaaffects plant growth by auxin production. Physiology Plantarum, 131, 581-589. https://doi.org/10.1111/j.1399-3054.2007.00983.x
36. Sun, , Johnson, J.M., Cai, D., Sherameti, I., Oelmüller, R., & Lou, B. (2010). Piriformospora indica confers drought tolerance in Chinese cabbage leaves by stimulating antioxidant enzymes, the expression of drought- related genes and the plastid-localized CAS protein. Journal of Plant Physiology, 167(12), 1009-1017. https://doi.org/10.1016/j.jplph.2010.02.013
37. Tebuqin Bao, Y.Y. (2015). Colonization characteristics of AMF in common mongolian medicinal plants of horqin sandy land. Inner Mongolia Agricultural Science and Technology, 43(6), 25-28. https://doi.org/10.3969/j.issn.1007-0907.2015.06.008
38. Ucar, , Ozyigit, Y., & Turgut, K. (2016). The effects of light and temperature on germination of stevia (Stevia rebaudiana BERT.) seeds. Türkiye Tarımsal Araştırmalar Dergisi, 3(1), 37-40.
39. Valinezhad, , Gholizadeh, A., Naeemi, M., Gholamalalipour Alamdari, E., & Zarei, M. (2019). Effects of vermicompost and mycorrhizal fungus on quantitative and qualitative traits of medicinal plant Stevia rebaudiana Bertoni. Iranian Journal of Medicinal and Aromatic Plants Research, 35(3), 484-500. (In Persian with English abstract). https://doi.org/10.22092/ijmapr.2019.123788.2417
40. Varma,, Sheramati, I., & Tripathi, S. (2012).The symbiotic fungus Piriformospora indica. Review. In Hock B (eds). Fungal Associations. 2nd Ed. Berlin: Springer; pp. 231-154.
41. Wu, S., Gao, W.Q., Srivastava, A.K., Zhang, F., & Zou, Y.N. (2020). Nutrient acquisition and fruit quality of Ponkan mandarin in response to AMF inoculation. Indian Journal of Agricultural Sciences, 90, 1563-1567.
42. Xu, , Tan, X., & Wang, Z. (2010). Effects of sucrose on germination and seedling development of Brassica Napus. International Journal of Biology, 2, 150-154.
43. Zare Hassanabadi, M Z., Dashti, M., & Akhondi, M. (2020). The effect of two species of Arbuscular mycorrhiza fungi on the activity of antioxidant enzymes and morphophysiological characteristics of Mentha pulegium in drought stress. Technology of Medicinal and Aromatic Plants of Iran, 2(2), 83-99. (In Persian with English abstract). https://doi.org/10.22092/MPT.2020.127803.1049
44. Zhang, H., Sun, J.Q., & Bao, Y.Y. (2015). Advances in studies on plant secondary metabolites influenced by Arbuscular mycorrhizal Journal of Agricultural Biotechnology, 23(8), 1093-1103. https://doi.org/10.3969/j.issn.1674-7968.2015.08.013
45. Zhang, , Proenca, R., & Maffei, M. (1994). Positional cloning of the mouse obese gene and its human homologue. Nature, 372, 425–432.
46. Zhang, Z., Zhang, , & Huang, Y. (2014). Effects of Arbuscular mycorrhizal fungi on the drought tolerance of Cyclobalanopsis glauca seedlings under greenhouse conditions. New For, 45, 545–556.