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Developing polyploid genetic resources for enhanced rebaudioside A synthesis and agronomic traits in Stevia rebaudiana Bertoni

Published online by Cambridge University Press:  27 December 2024

Sivendra Joshi
Affiliation:
CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Centre, Pantnagar, Udham Singh Nagar, Uttarakhand-263149, India
Venkatesha K. T.*
Affiliation:
CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Centre, Pantnagar, Udham Singh Nagar, Uttarakhand-263149, India
Abhilasha Gupta
Affiliation:
CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Centre, Pantnagar, Udham Singh Nagar, Uttarakhand-263149, India
Rajendra Chandra Padalia
Affiliation:
CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Centre, Pantnagar, Udham Singh Nagar, Uttarakhand-263149, India
Dipender Kumar
Affiliation:
CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Centre, Pantnagar, Udham Singh Nagar, Uttarakhand-263149, India
*
Corresponding author: Venkatesha K. T.; Email: venkatesha@cimap.res.in

Abstract

Stevia rebaudiana Bertoni stores over 20 glycosides in leaves and stem, the primary sweetening compounds are stevioside and rebaudioside A. Stevioside is 250–300 times and rebaudioside A is 350–450 times sweeter than sucrose. These glycosides can be substituted for free sugars being used in food and beverage industries. Due to limitations in the improvement of stevia through conventional breeding, induced polyploidy technique is a quick way to achieve enhanced leaf yield with increased secondary metabolites production. Presented study was conducted to develop genetic resources by inducing polyploidy and characterizing it. The apical meristem of 30-day-old seedlings was treated with colchicine solutions, ranging from 0.05 to 0.30%. Flow-cytometric analysis confirmed 13 tetraploids and one triploid genotype. Tetraploid plants showed significant changes in plant height, had larger leaf area, thicker leaves and higher levels of stevioside and rebaudioside A content, as compared to control. The triploid plant also surpassed the diploid control in number of branches per plant, leaf thickness, leaf area and fresh stem weight. The tetraploid line (ST 6) had 65.7% higher rebaudioside A content compared to diploid control. Correlation study revealed that leaf area and leaf thickness had an inverse correlation with rebaudioside A content. These newly developed polyploid genotypes offer valuable genetic resources for future stevia breeding programmes and as polyploid varieties for commercial cultivation.

Type
Research Article
Copyright
Copyright © The Author(s), 2024. Published by Cambridge University Press on behalf of National Institute of Agricultural Botany

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References

Abbas Momtazi-Borojeni, A, Esmaeili, SA, Abdollahi, E and Sahebkar, A (2017) A review on the pharmacology and toxicology of steviol glycosides extracted from Stevia rebaudiana. Current Pharmaceutical Design 23, 16161622.CrossRefGoogle Scholar
Adabiyah, R, Ratnadewi, D, Ermayanti, TM, Al Hafiizh, E and Susanti, EM (2023) Morphological and anatomical comparison between tetraploid Stevia rebaudiana Bertoni and its Parental Diploid. Hayati Journal of Biosciences 30, 321335.CrossRefGoogle Scholar
Ameer, K, Jiang, GH, Amir, RM and Eun, JB (2020) Antioxidant potential of Stevia rebaudiana Bertoni. In Victor, RP (ed.), Pathology. Cambridge, MA, United States: Academic Press, pp. 345356.CrossRefGoogle Scholar
Amien, S, Maulana, H, Ruswandi, D and Nurjanah, S (2021) Genetic gain and relationship of yield and yield attributes of mutant and cross-bred stevia (Stevia rebaudiana) genotypes. Biodiversitas: Journal of Biological Diversity 22, 31193126.CrossRefGoogle Scholar
Amin, K, Ozgen, S and Selamoglu, Z (2017) Stevia rebaudiana: a potential boon for human health. SM Journal of Medicinal Plant Studies 1, 1005.CrossRefGoogle Scholar
Azrul-Murad, AA, Yong, CS, Tan, YLAB and Ghani, NI (2022) Identification and characterization of genic simple sequence repeats from the leaf and stem transcriptomes of Stevia rebaudiana Bertoni. Scientia Horticulturae 300, 111067.CrossRefGoogle Scholar
Baghyalakshmi, K, Shaik, M, Mohanrao, MD, Shaw, RK, Lavanya, C, Manjunatha, T and Senthilvel, S (2020) Development and characterization of tetraploid castor plants. Plant Genetic Resources: Characterisation and Utilisation 18, 98104.CrossRefGoogle Scholar
Bertioli, DJ, Seijo, G, Freitas, FO, Valls, JFM, Leal-Bertioli, SCM and Moretzsohn, MC (2011) An overview of peanut and its wild relatives. Plant Genetic Resources: Characterisation and Utilisation 9, 134149.CrossRefGoogle Scholar
Blakeslee, AF and Avery, AG (1937) Methods of inducing doubling of chromosomes in plants: by treatment with colchicine. Journal of Heredity 28, 393411.CrossRefGoogle Scholar
Bovanová, L, Brandšteterová, E and Baxa, S (1998) HPLC determination of stevioside in plant material and food samples. Zeitschrift fur Lebensmittel -Untersuchung und -Forschung 207, 352355.Google Scholar
Brandle, J (1999) Genetic control of rebaudioside A and C concentration in leaves of the sweet herb, Stevia rebaudiana. Canadian Journal of Plant Science 79, 8591.CrossRefGoogle Scholar
Dhange, PR, Srinatha, TN and Kandpal, K (2023) Stevia farming: a moneymaking venture for farmers. Indian Farming 73, 1922.Google Scholar
Doležel, J, Greilhuber, J and Suda, J (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nature Protocols 2, 22332244.CrossRefGoogle ScholarPubMed
Grad, WE and Gomaa, SE (2020) Polyploidy and sweetener induction in different stevia varieties using colchicine. Egyptian Journal of Plant Breeding 4, 697715.Google Scholar
Hajihashemi, S and Ehsanpour, AA (2014) Antioxidant response of Stevia rebaudiana B. to polyethylene glycol and paclobutrazol treatments under in vitro culture. Applied Biochemistry and Biotechnology 172, 40384052.CrossRefGoogle Scholar
Hodkinson, TR, Klaas, M, Jones, MB, Prickett, R and Barth, S (2015) Miscanthus: a case study for the utilization of natural genetic variation. Plant Genetic Resources: Characterisation and Utilisation 13, 219237.CrossRefGoogle Scholar
Huber, BM and Wehner, TC (2023) Heritability and genetic variance estimates for agronomic traits and glycoside yield in four elite stevia breeding populations. Crop Science 63, 12341245.CrossRefGoogle Scholar
IBM Corp (2017) IBM SPSS Statistics for Windows, Version 25.0. Armonk, NY: IBM Corp.Google Scholar
Kaensaksiri, T, Soontornchainaksaeng, P, Soonthornchareonnon, N and Prathanturarug, S (2011) In vitro induction of polyploidy in Centella asiatica (L.) urban. Plant Cell, Tissue and Organ Culture 107, 187194.CrossRefGoogle Scholar
Kobayashi, N, Yamashit, S, Ohta, K and Hosoki, T (2008) Morphological characteristics and their inheritance in colchicine-induced Salvia polyploids. Journal of the Japanese Society for Horticultural Science 77, 186191.CrossRefGoogle Scholar
Le Bihan, Z, Cosson, P, Rolin, D and Schurdi-Levraud, V (2020) Phenological growth stages of stevia (Stevia rebaudiana Bertoni) according to the Biologische Bundesanstalt Bundessortenamt and Chemical Industry (BBCH) scale. Annals of Applied Biology 177, 404416.CrossRefGoogle Scholar
Li, H, Liu, X, Zhang, M, Feng, Z, Liu, D, Ayliffe, M, Hao, M, Ning, S, Yuan, Z, Yan, Z, Chen, X and Zhang, L (2018) Development and identification of new synthetic T. turgidum-T. monococcum amphiploids. Plant Genetic Resources: Characterisation and Utilisation 16, 555563.CrossRefGoogle Scholar
Manzoor, A, Ahmad, T, Bashir, MA, Baig, MMQ, Quresh, AA, Shah, MKN and Hafiz, IA (2018) Induction and identification of colchicine induced polyploidy in Gladiolus grandiflorus ‘White Prosperity’. Folia Horticulturae 30, 307319.CrossRefGoogle Scholar
Mishra, BK, Pathak, S, Sharma, A, Trivedi, PK and Shukla, S (2010) Modulated gene expression in newly synthesized auto-tetraploid of Papaver somniferum L. South African Journal of Botany 76, 447452.CrossRefGoogle Scholar
Miyagawa, H, Fujikowa, N, Kohda, H, Yamasaki, K, Taniguchi, K and Tanaka, R (1986) Studies on the tissue culture of Stevia rebaudiana and its components: (II). Induction of shoot primordia. Planta Medica 52, 321324.CrossRefGoogle Scholar
Mohd-Radzman, NH, Ismail, WIW, Adam, Z, Jaapar, SS and Adam, A (2013) Potential roles of Stevia rebaudiana Bertoni in abrogating insulin resistance and diabetes: a review. Evidence-Based Complementary and Alternative Medicine 2013, 718049.CrossRefGoogle ScholarPubMed
Nasirvand, S, Zakaria, RA, Zare, N and Esmaeilpoor, B (2018) Polyploidy induction in parsley (Petroselinum crispum L.) by colchicine treatment. Cytologia 83, 393396.CrossRefGoogle Scholar
Oddone, B (1997) How to Grow Stevia. Technical Manual. Pawtucket, CT: Guarani Botanicals.Google Scholar
Parris, CA, Shock, CC and Qian, M (2017) Soil water tension irrigation criteria affects Stevia rebaudiana leaf yield and leaf steviol glycoside composition. Horticultural Science 52, 154161.Google Scholar
Putnik, P, Bezuk, I, Barba, FJ, Lorenzo, JM, Polunić, I and Bursać, DK (2020) Sugar reduction: Stevia rebaudiana Bertoni as a natural sweetener. In F. J., Barba, P., Putnik and D., Bursać Kovačević (eds), Agri-food Industry Strategies for Healthy Diets and Sustainability. USA: Academic Press, pp. 123152.CrossRefGoogle Scholar
Qiu, LJ, Chen, PY, Liu, ZX, Li, YH, Guan, RX, Wang, LH and Chang, RZ (2011) The worldwide utilization of the Chinese soybean germplasm collection. Plant Genetic Resources: Characterisation and Utilisation 9, 109122.CrossRefGoogle Scholar
Ramesh, K, Sing, V and Megeji, NW (2006) Cultivation of stevia Stevia rebaudiana Bertoni: a comprehensive review. Advances in Agronomy 89, 137177.CrossRefGoogle Scholar
Ratner, B (2009) The correlation coefficient: its values range between +1/−1, or do they? Journal of Targeting, Measurement and Analysis for Marketing 17, 139142.CrossRefGoogle Scholar
R Core Team (2021) R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing. Available at https://www.R-project.orgGoogle Scholar
Sadat, NSA, Norouzi, M, Karimzadeh, G, Shirkool, K and Niazian, M (2017) Effect of colchicine-induced polyploidy on morphological characteristics and essential oil composition of ajowan (Trachyspermum ammi L.). Plant Cell, Tissue and Organ Culture 130, 543551.CrossRefGoogle Scholar
Soejima, A, Tanabe, AS, Takayama, I, Kawahara, T, Watanabe, K, Nakazawa, M, Mishima, M and Yahara, T (2017) Phylogeny and biogeography of the genus Stevia (Asteraceae: Eupatorieae): an example of diversification in the Asteraceae in the new world. Journal of Plant Research 130, 953972.CrossRefGoogle ScholarPubMed
Tripathi, S, Jadaun, JS, Chandra, M and Sangwan, NS (2016) Medicinal plant transcriptomes: the new gateways for accelerated understanding of plant secondary metabolism. Plant Genetic Resources: Characterisation and Utilisation 14, 256269.CrossRefGoogle Scholar
Wang, L and Wu, W (2019) Angiotensin-converting enzyme inhibiting ability of ethanol extracts, steviol glycosides and protein hydrolysates from stevia leaves. Food and Function 10, 79677972.CrossRefGoogle ScholarPubMed
World Health Organization (2015) Guideline: Sugars Intake for Adults and Children. Geneva, Switzerland: World Health Organization.Google Scholar
Xie, N, Zhao, Y, Huang, M, Chen, C, Cao, C, Wang, J, Shi, Z and Gao, J (2024) Polyploid induction and identification of Begonia × benariensis. Horticulturae 10, 47.CrossRefGoogle Scholar
Xu, X, Yuan, H, Yu, X, Huang, S, Sun, Y, Zhang, T, Liu, Q, Tong, H, Zhang, Y, Wang, Y, Liu, C, Wu, L, Hou, M and Yang, Y (2021) The chromosome-level Stevia genome provides insights into steviol glycoside biosynthesis. Horticulture Research 8, 110.CrossRefGoogle ScholarPubMed
Yadav, AK, Singh, S, Dhyani, D and Ahuja, PS (2011) A review on the improvement of stevia [Stevia rebaudiana (Bertoni)]. Canadian Journal of Plant Science 91, 127.CrossRefGoogle Scholar
Ye, YM, Tong, J, Shi, XP, Yuan, W and Li, GR (2010) Morphological and cytological studies of diploid and colchicine-induced tetraploid lines of crape myrtle (Lagerstroemia indica L.). Scientia Horticulturae 124, 95101.CrossRefGoogle Scholar
Yücesan, B, Mohammed, A, Büyükgöçmen, R, Altuğ, C, Kavas, Ö, Gürel, S and Gürel, E (2016) In vitro and ex vitro propagation of Stevia rebaudiana Bertoni with high Rebaudioside A content – A commercial scale application. Scientia Horticulturae 203, 2028.CrossRefGoogle Scholar
Zhang, H, An, S, Hu, J, Lin, Z, Liu, X, Bao, H and Chen, R (2018) Induction, identification and characterization of polyploidy in Stevia rebaudiana Bertoni. Plant Biotechnology 35, 8186.CrossRefGoogle ScholarPubMed
Zhou, J, Guo, F, Fu, J, Xiao, Y and Wu, J (2020) In vitro polyploid induction using colchicine for Zingiber officinale Roscoe cv ‘Fengtou’ ginger. Plant Cell, Tissue and Organ Culture 142, 8794.CrossRefGoogle Scholar
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