Potassium deficiency inhibits steviol glycosides synthesis by limiting leaf sugar metabolism in stevia (Stevia rebaudiana Bertoni) plants

doi:10.1016/S2095-3119(20)63472-4

摘要/Abstract

摘要：

甜菊叶片中的甜菊糖苷由于具有高甜度和低热量的特点而凸显价值，这推动了甜叶菊商业化种植的发展。优化施肥管理可以有效提高甜菊糖苷的生产力，但是目前关于钾肥与甜菊糖苷生产之间的关系尚不明确。本研究通过盆栽试验揭示了甜菊缺钾对叶片甜菊糖苷合成的影响并探索了其潜在机制。结果表明，在高钾土壤背景下，与常规施钾相比，不施钾肥对甜菊生物量没有显著影响。然而，不施钾肥显著降低了叶片中甜菊糖苷的含量以及甜菊糖苷合成相关基因的表达水平。在缺钾条件下，叶片中不同糖组分含量显著降低，糖代谢相关酶的活性受到抑制。此外，通过对甜菊幼苗叶片进行蔗糖喷施可以有效减弱缺钾造成的甜菊糖苷抑制作用。研究结果还揭示了甜菊叶片中蔗糖、葡萄糖与甜菊糖苷含量之间的显著正相关关系。综上所述，我们的研究结果表明缺钾会抑制甜菊叶片中甜菊糖苷合成，而这种抑制作用是由叶片糖代谢介导的。我们的发现为进一步提高甜菊糖苷的生产潜力提供了新的见解。

Abstract:

The steviol glycosides (SGs) in stevia (Stevia rebaudiana Bertoni) leaves are becoming increasingly valuable due to its high sweetness but low calorific value, which is driving the development of stevia commercial cultivation. Optimizing fertilization management can effectively increase SGs productivity, but knowledge on the relationship between potassium (K) fertilization and SGs production is still lacking. In this study, pot experiments were conducted in order to investigate the effect of K deficiency on SGs synthesis in stevia leaves, as well as the underlying mechanisms. Our results showed that when compared with standard K fertilization, K deficiency treatment has no significant effect on the biomass of stevia plant grown in a given soil with high K contents. However, K deficiency critically decreased leaf SGs contents as well as the expression of SGs synthesis-related genes. The contents of different sugar components decreased and the activities of sugar metabolism-related enzymes were inhibited under the K deficiency condition. Moreover, spraying sucrose on the leaves of stevia seedlings diminished the inhibitory effect caused by K deficiency. Our results also revealed the significant positive correlations between sucrose, glucose and SGs contents. Overall, our results suggest that K deficiency would suppress the synthesis of SGs in stevia leaves, and this effect may be mediated by the leaf sugar metabolism. Our findings provide new insights into the improvement of SGs production potential.

Key words: Stevia rebaudiana Bertoni , steviol glycosides , potassium deficiency , sugar metabolism

. [J]. Journal of Integrative Agriculture, 2021, 20(11): 2932-2943.

SUN Yu-ming, HUANG Xiao-lei, ZHANG Ting, YANG Yong-heng, CHENG Xiao-fang, XU Xiao-yang, YUAN Hai-yan. Potassium deficiency inhibits steviol glycosides synthesis by limiting leaf sugar metabolism in stevia (Stevia rebaudiana Bertoni) plants[J]. Journal of Integrative Agriculture, 2021, 20(11): 2932-2943.

参考文献

Aghighi S M, Omidi H, Tabatabaei S J. 2019. Stevia (Stevia rebaudiana Bertoni) responses to NaCl stress: Growth, photosynthetic pigments, diterpene glycosides and ion content in root and shoot. Journal of the Saudi Society of Agricultural Sciences, 18, 355–360.
Angelini L G, Martini A, Passera B, Tavarini S. 2016. Cultivation of Stevia Rebaudiana Bertoni and Associated Challenges. In: Mérillon J M, Ramawat K G, eds., Sweeteners.Springer International Publishing, Switzerland. pp. 1–52.
Armengaud P, Sulpice R, Miller A J, Stitt M, Amtmann A, Gibon Y. 2009. Multilevel analysis of primary metabolism provides new insights into the role of potassium nutrition for glycolysis and nitrogen assimilation in Arabidopsis roots. Plant Physiology, 150, 772–785.
Arsenault P R, Vail D R, Wobbe K K, Weathers P J. 2010. Effect of sugars on artemisinin production in Artemisia annua L.: Transcription and metabolite measurements. Molecules, 15, 2302–2318.
Barbet-Massin C, Giuliano S, Alletto L, Dayde J, Berger M. 2015. Nitrogen limitation alters biomass production but enhances steviol glycoside concentration in Stevia rebaudiana bertoni. PLoS ONE, 10, e0133067.
Barickman T C, Horgan T E, Wheeler J R, Sams C E. 2016. Elevated levels of potassium in greenhouse-grown red romaine lettuce impacts mineral nutrient and soluble sugar concentrations. HortScience, 51, 504–509.
Bondarev N, Reshetnyak O, Nosov A. 2003. Effects of nutrient medium composition on development of Stevia rebaudiana shoots cultivated in the roller bioreactor and their production of steviol glycosides. Plant Science, 165, 845–850.
Brunelle T, Dumas P, Souty F, Dorin B, Nadaud F. 2015. Evaluating the impact of rising fertilizer prices on crop yields. Agricultural Economics, 46, 653–666.
Cantabella D, Piqueras A, Acosta-Motos J R, Bernal-Vicente A, Hernández J A, Díaz-Vivancos P. 2017. Salt-tolerance mechanisms induced in Stevia rebaudiana Bertoni: Effects on mineral nutrition, antioxidative metabolism and steviol glycoside content. Plant Physiology and Biochemistry, 115, 484–496.
Ceunen S, Geuns J. 2013a. Glucose, sucrose, and steviol glycoside accumulation in Stevia rebaudiana grown under different photoperiods. Biologia Plantarum, 57, 390–394.
Ceunen S, Geuns J. 2013b. Influence of photoperiodism on the spatio-temporal accumulation of steviol glycosides in Stevia rebaudiana (Bertoni). Plant Science, 198, 72–82.
Chrysargyris A, Drouza C, Tzortzakis N. 2017a. Optimization of potassium fertilization/nutrition for growth, physiological development, essential oil composition and antioxidant activity of Lavandula angustifolia Mill. Journal of Soil Science and Plant Nutrition, 17, 291–306.
Chrysargyris A, Xylia P, Botsaris G, Tzortzakis N. 2017b. Antioxidant and antibacterial activities, mineral and essential oil composition of spearmint (Mentha spicata L.) affected by the potassium levels. Industrial Crops and Products, 103, 202–212.
Esetlili B Ç, Öztürk B, Çobano?lu Ö, Anaç D. 2015. Sweet basil (Ocimum basilicum L.) and potassium fertilization. Journal of Plant Nutrition, 39, 35–44.
Ghorbani T, Kahrizi D, Saeidi M, Arji I. 2017. Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cellular and Molecular Biology, 63, 33–37.
Guo R, Yan H, Li X, Zou X, Zhang X, Yu X, Ci D, Wang Y, Si T. 2020. Green leaf volatile (Z)-3-hexeny-1-yl acetate reduces salt stress in peanut by affecting photosynthesis and cellular redox homeostasis. Physiologia Plantarum, 170, 75–92.
Hafsi M, Pfeiffer W, Monneveux P. 2003. Flag leaf senescence, carbon content and carbon isotope discrimination in durum wheat grown under semi-arid conditions. Cereal Research Communications, 31, 161–168.
Heidari S, Azizi M, Soltani F, Hadian J. 2014. Foliar application of Ca(NO3)2 and KNO3 affects growth, essential oil content, and oil composition of French tarragon. Industrial Crops and Products, 62, 526–532.
Hu W, Coomer T D, Loka D A, Oosterhuis D M, Zhou Z. 2017. Potassium deficiency affects the carbon–nitrogen balance in cotton leaves. Plant Physiology and Biochemistry, 115, 408–417.
Hu W, Dai Z, Yang J, Snider J L, Wang S, Chen B, Zhou Z. 2018a. The variability of cottonseed yield under different potassium levels is associated with the changed oil metabolism in embryo. Field Crops Research, 224, 80–90.
Hu W, Loka D A, Fitzsimons T R, Zhou Z, Oosterhuis D M. 2018b. Potassium deficiency limits reproductive success by altering carbohydrate and protein balances in cotton (Gossypium hirsutum L.). Environmental and Experimental Botany, 145, 87–94.
Ibrahim M, Jaafar H, Karimi E, Ghasemzadeh A. 2012. Primary, secondary metabolites, photosynthetic capacity and antioxidant activity of the malaysian herb kacip fatimah (Labisia Pumila Benth) exposed to potassium fertilization under greenhouse conditions. International Journal of Molecular Sciences, 13, 15321–15342.
Karimi M, Ahmadi A, Hashemi J, Abbasi A, Tavarini S, Guglielminetti L, Angelini L G. 2015. The effect of soil moisture depletion on Stevia (Stevia rebaudiana Bertoni) grown in greenhouse conditions: Growth, steviol glycosides content, soluble sugars and total antioxidant capacity. Scientia Horticulturae, 183, 93–99.
Karimi M, Moradi K. 2018. The response of Stevia (Stevia rebaudiana Bertoni) to nitrogen supply under greenhouse condition. Journal of Plant Nutrition, 41, 1695–1704.
Lemus-Mondaca R, Vega-Galvez 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, 1121–1132.
Liu W, Zhu D, Liu D, Zhou W, Yang T, Geng M. 2011. Influence of potassium deficiency on flower yield and flavonoid metabolism in leaves of Chrysanthemum morifolium Ramat. Journal of Plant Nutrition, 34, 1905–1918.
Lu L, He C, Jin Y, Zhang X, Wei J. 2013. Effects of the applications of phosphorus and potassium fertilizers at different growth stages on the root growth and bioactive compounds of ‘Salvia miltiorrhiza’ Bunge. Australian Journal of Crop Science, 7, 1533.
Lu Z, Lu J, Pan Y, Lu P, Li X, Cong R, Ren T. 2016. Anatomical variation of mesophyll conductance under potassium deficiency has a vital role in determining leaf photosynthesis. Plant Cell and Environment, 39, 2428–2439.
Ma L, Ren G X, Shi Y. 2011. Effects of potassium fertilizer on diurnal change of photosynthesis in Stevia Rebaudiana Bertoni. Advanced Materials Research, 343, 1087–1091.
Ma L, Shi Y. 2011. Effects of potassium fertilizer on physiological and biochemical index of Stevia rebaudiana Bertoni. Energy Procedia, 5, 581–586.
Nelson D, Sommers L. 1972. A simple digestion procedure for estimation of total nitrogen in soils and sediments. Journal of Environmental Quality, 1, 423–425.
Pal P K, Kumar R, Guleria V, Mahajan M, Prasad R, Pathania V, Gill B S, Singh D, Chand G, Singh B, Singh R D, Ahuja P S. 2015. Crop-ecology and nutritional variability influence growth and secondary metabolites of Stevia rebaudiana Bertoni. BMC Plant Biology, 15, 67.
Prakash I, Markosyan A, Bunders C. 2014. Development of next generation stevia sweetener: Rebaudioside M. Foods, 3, 162–175.
Ramesh K, Singh V, Megeji N W. 2006. Cultivation of stevia Stevia rebaudiana (Bert.) Bertoni: A comprehensive review. Advances in Agronomy, 89, 137–177.
Ritu M, Nandini J. 2016. Nutritional composition of Stevia rebaudiana, a sweet herb, and its hypoglycaemic and hypolipidaemic effect on patients with non-insulin dependent diabetes mellitus. Journal of the Science of Food and Agriculture, 96, 4231–4234.
Römheld V, Kirkby E A. 2010. Research on potassium in agriculture: Needs and prospects. Plant and Soil, 335, 155–180.
Santos G, Guerrero C, Reis M, Miguel G, Kienle U. 2000. Stevioside and rebaudioside A foliar leaf content changes in plants of Stevia rebaudiana Bertoni as a response to the potassium fertilization. In: Proceedings of the 19e Journées Internationales Huiles Essentiales et Extraits. 30 August–2 September, Promotion des Plantes à Parfum, Aromatiques et Médicinales (APPAM). Digne les Bains, França. pp. 553–560.
Shaw T M, Moore J A, Marshall J D. 1998. Root chemistry of douglas-fir seedlings grown under different nitrogen and potassium regimes. Canadian Journal of Forest Research, 28, 1566–1573.
Shen C, Shi X, Xie C, Li Y, Yang H, Mei X, Xu Y, Dong C. 2019. The change in microstructure of petioles and peduncles and transporter gene expression by potassium influences the distribution of nutrients and sugars in pear leaves and fruit. Journal of Plant Physiology, 232, 320–333.
Shen C, Wang J, Shi X, Kang Y, Xie C, Peng L, Dong C, Shen Q, Xu Y. 2017. Transcriptome analysis of differentially expressed genes induced by low and high potassium levels provides insight into fruit sugar metabolism of pear. Frontiers in Plant Science, 8, 938.
Sun Y, Guo J, Li Y, Luo G, Li L, Yuan H, Mur L A J, Guo S. 2020a. Negative effects of the simulated nitrogen deposition on plant phenolic metabolism: A meta-analysis. The Science of the Total Environment, 719, 137442.
Sun Y, Hou M, Mur L A J, Yang Y, Zhang T, Xu X, Huang S, Tong H. 2019. Nitrogen drives plant growth to the detriment of leaf sugar and steviol glycosides metabolisms in Stevia (Stevia rebaudiana Bertoni). Plant Physiology and Biochemistry, 141, 240–249.
Sun Y, Yang Y, Hou M, Huang X, Zhang T, Huang S, Xu X, Yuan H. 2020b. Optimized nitrogen topdressing strategies enhance steviol glycoside productivity in stevia (Stevia rebaudiana Bertoni) plants. Journal of Soil Science and Plant Nutrition, 20, 1133–1143.
Sung J, Lee S, Lee Y, Ha S, Song B, Kim T, Waters B M, Krishnan H B. 2015. Metabolomic profiling from leaves and roots of tomato (Solanum lycopersicum L.) plants grown under nitrogen, phosphorus or potassium-deficient condition. Plant Science, 241, 55–64.
Taber H, Perkins-Veazie P, Li S, White W, Rodermel S, Xu Y. 2008. Enhancement of tomato fruit lycopene by potassium is cultivar dependent. HortScience, 43, 159–165.
Tavarini S, Pagano I, Guidi L, Angelini L G. 2016. Impact of nitrogen supply on growth, steviol glycosides and photosynthesis in Stevia rebaudiana Bertoni. Plant Biosystems, 150, 953–962.
Tavarini S, Passera B, Angelini L G. 2018. Crop and steviol glycoside improvement in Stevia by breeding. In: Steviol Glycosides: Cultivation, Processing, Analysis and Applications in Food. Royal Society of Chemistry, London. pp. 1–31.
Tian S, Guo R, Zou X, Zhang X, Yu X, Zhan Y, Ci D, Wang M, Wang Y, Si T. 2019. Priming with the green leaf volatile (Z)-3-Hexeny-1-yl acetate enhances salinity stress tolerance in peanut (Arachis hypogaea L.) seedlings. Frontiers in Plant Science, 10, 785.
Vasilakoglou I, Kalfountzos D, Gougoulias N, Reppas C. 2016. Productivity of two stevia varieties under reduced irrigation and fertilization inputs. Archives of Agronomy and Soil Science, 62, 457–472.
Wang J, Li S, Xiong Z, Wang Y. 2016. Pathway mining-based integration of critical enzyme parts for de novo biosynthesis of steviolglycosides sweetener in Escherichia coli. Cell Research, 26, 258.
Wang J, Zhao H, Wang Y, Lau H, Zhou W, Chen C, Tan S. 2020. A review of stevia as a potential healthcare product: Up-to-date functional characteristics, administrative standards and engineering techniques. Trends in Food Science and Technology, 103, 264–281.
Wang M, Zheng Q, Shen Q, Guo S. 2013. The critical role of potassium in plant stress response. International Journal of Molecular Sciences, 14, 7370–7390.
Wang Y, Weathers P. 2007. Sugars proportionately affect artemisinin production. Plant Cell Reports, 26, 1073–1081.
Webb H, Lanfear R, Hamill J, Foley W J, Külheim C. 2013. The yield of essential oils in Melaleuca alternifolia (Myrtaceae) is regulated through transcript abundance of genes in the MEP pathway. PLoS ONE, 8, 3.
Yadav S K, Guleria P. 2012. Steviol glycosides from stevia: Biosynthesis pathway review and their application in foods and medicine. Critical Rviews in Fod Science and Nutrition, 52, 988–998.
Zahoor R, Dong H, Abid M, Zhao W, Wang Y, Zhou Z. 2017. Potassium fertilizer improves drought stress alleviation potential in cotton by enhancing photosynthesis and carbohydrate metabolism. Environmental and Experimental Botany, 137, 73–83.
Zörb C, Senbayram M, Peiter E. 2014. Potassium in agriculture - status and perspectives. Journal of Plant Physiology, 171, 656–669.
Zhang T, Shankar A, Singh A, Kanwar P, Srivastava A K, Pandey A, Suprasanna P, Kapoor S, Pandey G K. 2013. Gene expression analysis of rice seedling under potassium deprivation reveals major changes in metabolism and signaling components. PLoS ONE, 8, e70321.
Zhang W, Zhang N, Zhao J, Guo Y, Zhao Z, Mei L. 2017. Potassium fertilization improves apple fruit (Malus domestica Borkh. cv. Fuji) development by regulating trehalose metabolism. The Journal of Horticultural Science and Biotechnology, 92, 539–549.