Issue
Effect of salinity on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions
Corresponding Author(s) : M. Ghaheri
Cellular and Molecular Biology,
Vol. 63 No. 7: Issue 7
Abstract
Stevia rebaudiana Bertoni is a famous medicinal plant for its low calorific value compounds which are named steviol glycosides (SGs) and they are 150-300 times sweeter than sugar. Among various SGs, stevioside and rebaudioside A considered to be the main sweetening compounds. Soil salinity is one of the most essential stress in the world. Salinity affects the survival and yield of crops. In current study the effects of salinity and osmotic stress caused by different concentration of NaCl (0, 20, 40, 60 and 80 mM) on morphological traits, genes expressionand amount of both stevioside and rebaudioside Aunder in vitro conditions has been investigated. The morphological traits such as bud numbers, root numbers, shoot length (after 15 and 30 days) were evaluated. With increasing salinity, the values of all studied morphological traits decreased. To investigation of UGT74G1 and UGT76G1 genes expression that are involved in the synthesis of SGs, RT-PCR was done and there were significant differences between all media. The highest expression of both genes was observed in plantlets grown on MS media (with NaCl-free). Also, the lowest amounts of gene expression of the both genes were seen in MS+ 60 mM NaCl. Based on HPLC results, the highest amount of both stevioside and rebaudioside A were observed in plantlets grown in MS media (with NaCl-free). Finally, it can be concluded that stevia can survive under salt stress, but it has the best performance in the lower salinity.
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- Akbari F, Arminian A, Kahrizi D, Fazeli A. Effect of nitrogen sources on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Cell Mol Biol 2017; 63: 2.
- Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist 2017; 11(2): (online first)
- Hajihashemi Sh, Ehsanpour AA. Antioxidant Response of Stevia rebaudiana B. to Polyethylene Glycol and Paclobutrazol Treatments Under In Vitro Culture. Appl Biochem Biotechnol 2014; 172: 4038–4052.
- Saifi M, Nasrullah N, Ahmad MM, Athar A, Khan JA, Abdin, MZ. In silico analysis and expression profiling of miRNAs targeting genes of steviol glycosides biosynthetic pathway and their relationship with steviol glycosides content in different tissues of Stevia rebaudiana. Plant Physiol Biochem, 2015; 94: 57-64.
- Pandey H, Pandey P, Pandey ShSh, Singh S, Banerjee S. Meeting the challenge of stevioside production in the hairy roots of Stevia rebaudiana by probing the underlying process. Plant Cell Tiss Organ Cult 2016; 126: 511–52.
- Geuns JMC. Stevioside. Phytochemistry 2003; 64: 913–921.
- Philippe RN, Mey MD, Anderson J, Ajilkumar PK. Biotechnological production of natural zero-calorie sweeteners. Curr Opin Biotechnol 2014; 26: 155–161.
- Esmaeilia F, Kahrizi D, Mansouri M, Yari Kh, Kazemi N, Ghaheri M. Cell Dedifferentiation in Stevia rebauiana as a Pharmaceutical and Medicinal Plant. J Rep Pharm. Sci 2016; 5(1), 12-17.
- Kumar H, Singh K, Kumar S. 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Stevia rebaudiana Bertoni is a functional gene. Mol Biol Rep 2012; 39(12):10971-8.
- Ahmad Khan Sh, Rahman L, Verma R, Shanker K. Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiol Plant 2016; 38: 4.
- Hajihashemi S, Ehsanpour AA. Influence of Exogenously Applied Paclobutrazol on Some Physiological Traits and Growth of Stevia rebaudiana under in Vitro Drought Stress. Biologia 2013; 68: 414-420.
- Brandle JE, Telmer PG. Steviol glycoside biosynthesis. Phytochemistry 2007; 68: 1855–1863.
- Guleria P, Kumar Yadav S. Identification of miR414 and Expression Analysis of Conserved miRNAs from Stevia rebaudiana. Genomics Proteomics Bioinformatics 2011; 9(6): 211-217.
- Islam MR, Singh RK, Salam MA, Hassan L, Gregorio GB. Molecular diversity of stress tolerant rice genotypes using SSR markers. SABRAO J Breed Gent 2014; 40(2): 127-139.
- Rathore Sh, Singh N, Singh SK. Influence of NaCl on Biochemical Parameters of Two Cultivars of Stevia rebaudiana Regenerated in vitro. J Stress Physiol Biochem 2014; 10(2): 287-296.
- Zeng J, Chen A, Li D, Yi B, Wu W. Effects of salt stress on the growth, physiological responses, and glycoside contents of Stevia rebaudiana Bertoni. J Agric Food Chem 2013; 61(24): 5720-5726.
- Pandey M, Chikara SK. In vitro regeneration and effect of abiotic stress on physiology and biochemical content of Stevia rebaudiana ‘Bertoni'. J Plant Sci Res 2014; 1(3): 113.
- Jamil M, Bashir S, Anwar S, Bibi S, Bangash A, Ullah F, Shikrha E. Effect of salinity on physiological and biochemical characteristics of different varieties of rice. Pak J Biol Sci 2012; 44: 7-
- Kamran Khan M, Misra P, Sharma P, Shukla PK, Ramteke PW. Effect of adenine sulphate on in vitro mass propagation of Stevia rebaudiana Bertoni. J Med Plants Res 2014; 8(13): 543-549.
- Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an Anti-diabetic Plant - Stevia rebaudiana Bertoni, (Natural Citation as Sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA J Biol Sci 2012; 1(3): 37-42.
- El-Housini EA, Ahmed MA, Hassanein MS, Tawfik MM. Effect of Salicylic Acid (SA) on Growth and Quality of Stevia (Stevia rebaudiana Bert.) Under Salt Stress. American-Eurasian J Agric Environ Sci 2015; 14: 275-281.
- Gupta P, Sharma S, Saxena S. Effect of Salts (NaCl and Na2CO3) on Callus and Suspension Culture of Stevia rebaudiana for Steviol glycoside Production. Appl Biochem Biotechnol 2014; 172: 2894–2906.
- Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15: 473–479.
- Marone M, Mozzetti S, Ritis DD, Pierelli L, Scambia G. Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 2001; 3(1): 19-25.
- Jaitak V, Singh BB, Kaul VK. An efficient microwave-assisted extraction process of stevioside and rebaudioside-A from Stevia rebaudiana (Bertoni). Phytochem Anal 2009; 20: 240–245.
- Gupta P, Sharma S, Saxena S. Effect of abiotic stress on growth parameters and steviol glycoside content in Stevia rebaudiana (Bertoni) raised in vitro. J Appl Res Med Aromat Plants 2016; 3(4): 160-167.
References
Akbari F, Arminian A, Kahrizi D, Fazeli A. Effect of nitrogen sources on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Cell Mol Biol 2017; 63: 2.
Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist 2017; 11(2): (online first)
Hajihashemi Sh, Ehsanpour AA. Antioxidant Response of Stevia rebaudiana B. to Polyethylene Glycol and Paclobutrazol Treatments Under In Vitro Culture. Appl Biochem Biotechnol 2014; 172: 4038–4052.
Saifi M, Nasrullah N, Ahmad MM, Athar A, Khan JA, Abdin, MZ. In silico analysis and expression profiling of miRNAs targeting genes of steviol glycosides biosynthetic pathway and their relationship with steviol glycosides content in different tissues of Stevia rebaudiana. Plant Physiol Biochem, 2015; 94: 57-64.
Pandey H, Pandey P, Pandey ShSh, Singh S, Banerjee S. Meeting the challenge of stevioside production in the hairy roots of Stevia rebaudiana by probing the underlying process. Plant Cell Tiss Organ Cult 2016; 126: 511–52.
Geuns JMC. Stevioside. Phytochemistry 2003; 64: 913–921.
Philippe RN, Mey MD, Anderson J, Ajilkumar PK. Biotechnological production of natural zero-calorie sweeteners. Curr Opin Biotechnol 2014; 26: 155–161.
Esmaeilia F, Kahrizi D, Mansouri M, Yari Kh, Kazemi N, Ghaheri M. Cell Dedifferentiation in Stevia rebauiana as a Pharmaceutical and Medicinal Plant. J Rep Pharm. Sci 2016; 5(1), 12-17.
Kumar H, Singh K, Kumar S. 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Stevia rebaudiana Bertoni is a functional gene. Mol Biol Rep 2012; 39(12):10971-8.
Ahmad Khan Sh, Rahman L, Verma R, Shanker K. Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiol Plant 2016; 38: 4.
Hajihashemi S, Ehsanpour AA. Influence of Exogenously Applied Paclobutrazol on Some Physiological Traits and Growth of Stevia rebaudiana under in Vitro Drought Stress. Biologia 2013; 68: 414-420.
Brandle JE, Telmer PG. Steviol glycoside biosynthesis. Phytochemistry 2007; 68: 1855–1863.
Guleria P, Kumar Yadav S. Identification of miR414 and Expression Analysis of Conserved miRNAs from Stevia rebaudiana. Genomics Proteomics Bioinformatics 2011; 9(6): 211-217.
Islam MR, Singh RK, Salam MA, Hassan L, Gregorio GB. Molecular diversity of stress tolerant rice genotypes using SSR markers. SABRAO J Breed Gent 2014; 40(2): 127-139.
Rathore Sh, Singh N, Singh SK. Influence of NaCl on Biochemical Parameters of Two Cultivars of Stevia rebaudiana Regenerated in vitro. J Stress Physiol Biochem 2014; 10(2): 287-296.
Zeng J, Chen A, Li D, Yi B, Wu W. Effects of salt stress on the growth, physiological responses, and glycoside contents of Stevia rebaudiana Bertoni. J Agric Food Chem 2013; 61(24): 5720-5726.
Pandey M, Chikara SK. In vitro regeneration and effect of abiotic stress on physiology and biochemical content of Stevia rebaudiana ‘Bertoni'. J Plant Sci Res 2014; 1(3): 113.
Jamil M, Bashir S, Anwar S, Bibi S, Bangash A, Ullah F, Shikrha E. Effect of salinity on physiological and biochemical characteristics of different varieties of rice. Pak J Biol Sci 2012; 44: 7-
Kamran Khan M, Misra P, Sharma P, Shukla PK, Ramteke PW. Effect of adenine sulphate on in vitro mass propagation of Stevia rebaudiana Bertoni. J Med Plants Res 2014; 8(13): 543-549.
Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an Anti-diabetic Plant - Stevia rebaudiana Bertoni, (Natural Citation as Sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA J Biol Sci 2012; 1(3): 37-42.
El-Housini EA, Ahmed MA, Hassanein MS, Tawfik MM. Effect of Salicylic Acid (SA) on Growth and Quality of Stevia (Stevia rebaudiana Bert.) Under Salt Stress. American-Eurasian J Agric Environ Sci 2015; 14: 275-281.
Gupta P, Sharma S, Saxena S. Effect of Salts (NaCl and Na2CO3) on Callus and Suspension Culture of Stevia rebaudiana for Steviol glycoside Production. Appl Biochem Biotechnol 2014; 172: 2894–2906.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15: 473–479.
Marone M, Mozzetti S, Ritis DD, Pierelli L, Scambia G. Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 2001; 3(1): 19-25.
Jaitak V, Singh BB, Kaul VK. An efficient microwave-assisted extraction process of stevioside and rebaudioside-A from Stevia rebaudiana (Bertoni). Phytochem Anal 2009; 20: 240–245.
Gupta P, Sharma S, Saxena S. Effect of abiotic stress on growth parameters and steviol glycoside content in Stevia rebaudiana (Bertoni) raised in vitro. J Appl Res Med Aromat Plants 2016; 3(4): 160-167.