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Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities

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Abstract

The leaf explants of Stevia rebaudiana were exposed to a range of EMS (chemical mutagen) and gamma radiation (physical mutagen) doses. Among all the treatments, doses of 0.4 % v/v EMS and 0.95 KR gamma radiation were found to be most effective treatment for selection of variants via direct shoot bud induction. The field-established plants designated as EMS (E), gamma (G), and control (C) plants were subjected to photosynthetic parameter studies where E and G plants showed lower chlorophyll content, transpiration rate, CO2 exchange rate, and stomatal conductance. The implanted field soil at the time of harvesting of plants showed reduction in organic carbon (OC), electrical conductivity (EC), phosphorus (P)/potassium (K), and total Kjeldahl nitrogen (TKN). The ISSR profiling of E, G, and C scored total of 107 bands out of which 63 % were polymorphic bands, and plant E was found to be more distant from the C plant phylogenetically. On phytochemical analysis, the G plant registered twofold enhanced rebaudioside A with lower stevioside (3.2 ± 0.22 % dry wt.) content, whereas E plant showed more than 1.5- and 2.0-fold increase in stevioside and rebaudioside A as compared to control plants. Both the E (3.1 ± 0.15 % dry wt.) and G (2.3 ± 0.21 % dry wt.) plants registered lower steviol. The enhanced steviol glycoside profile was supported by the RT-PCR analysis of UGT74G1 and UGT76G1 that corresponds to stevioside and rebaudioside A biosynthesis, respectively. While the E plant showed 5–6-fold increase in the UGT74G1 (RQ = 5.51 ± 0.5) and UGT76G1 (RQ = 6.61 ± 0.5) gene expression, the G plant showed 5-fold increase in UGT76G1 (RQ = 5.29 ± 0.2) gene expression.

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Acknowledgments

We gratefully acknowledge the financial support received from the Council of Scientific & Industrial Research as CSIR-SRF, Government of India, and New Delhi to carry out this work and Director CIMAP for providing all the necessary facilities. We are highly thankful to Dr. NK Srivastava for helping in quantification of photosynthetic parameters.

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Authors and Affiliations

  1. Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India

    Shamshad Ahmad Khan & Laiq ur Rahman

  2. Department of Soil Analysis, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India

    Rajesh Verma

  3. Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Kukrail Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India

    Karuna Shanker

Authors
  1. Shamshad Ahmad Khan
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  2. Laiq ur Rahman
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  3. Rajesh Verma
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  4. Karuna Shanker
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Correspondence to Laiq ur Rahman.

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Communicated by M. H. Walter.

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Khan, S.A., Rahman, L.u., Verma, R. et al. Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiol Plant 38, 4 (2016). https://doi.org/10.1007/s11738-015-2003-8

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  • DOI: https://doi.org/10.1007/s11738-015-2003-8

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