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Steviol glycosides profile in Stevia rebaudiana Bertoni hairy roots cultured under oxidative stress-inducing conditions.
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2020-05-28 , DOI: 10.1007/s00253-020-10661-5
Marta Libik-Konieczny 1 , Żaneta Michalec-Warzecha 1 , Michał Dziurka 1 , Olga Zastawny 2 , Robert Konieczny 2 , Piotr Rozpądek 3 , Laura Pistelli 4
Affiliation  

The ability to synthesize particular steviol glycosides (SvGls) was studied in Stevia rebaudiana Bertoni hairy roots (HR) grown in the light or in the dark under the influence of different osmotic active compounds. Manipulation of culture conditions led to changes in the morphology and growth rate of HR, as well as to an increase in oxidative stress manifested as an enhancement in endogenous hydrogen peroxide concentration in the cultured samples. The highest level of H2O2 was noted in HR cultured under light or in the medium with the highest osmotic potential. This correlated with the highest increase in the expression level of ent-kaurenoic acid hydroxylase, responsible for the redirection of metabolic route to SvGls biosynthesis pathway. An analysis of transcriptional activity of some UDPglucosyltransferase (UGT85c2, UGT74g1, UGT76g1) revealed that all of them were upregulated due to the manipulation of culture conditions. However, the level of their upregulation depended on the type of stress factor used in our experiment. Analysis of SvGls content revealed that HR grown under all applied conditions were able to synthesize and accumulate several SvGls but their concentration differed between the samples across the different conditions. The level of rebaudioside A concentration exceeded the content of stevioside in HR in all tested conditions. Concomitantly, the presence of some minor SvGls, such as steviolbioside and rebaudioside F, was confirmed only in HR cultured in the lowest osmotic potential of the medium while rebaudioside D was also detected in the samples cultured in the media supplemented with NaCl or PEG.

Key Points
● Several steviol glycosides are synthesized in hairy roots of S. rebaudiana.
● Light or osmotic factors cause enhancement in oxidative stress level in hairy roots.
● It correlates with a significant increase in the level of KAH expression.
● UGTs expression and steviol glycosides content depends on culture conditions.


中文翻译:

在氧化应激诱导条件下培养的甜叶菊多毛根中的甜菊醇糖苷谱。

研究了在不同的渗透活性化合物的影响下,在明亮或黑暗条件下生长的甜叶菊甜叶菊毛状根(HR)中合成特定甜菊醇糖苷(SvGls)的能力。操纵培养条件会导致HR的形态和生长速率发生变化,并导致氧化应激的增加,这表现为培养样品中内源性过氧化氢浓度的增加。在光或具有最高渗透潜力的培养基中培养的HR中,H 2 O 2的含量最高。这与对氨基脲酸羟化酶表达水平的最高增加相关,负责将代谢途径重定向至SvGls生物合成途径。某些UDP葡萄糖基转移酶UGT85c2UGT74g1UGT76g1)的转录活性分析。)揭示,由于培养条件的操纵,所有这些蛋白均被上调。但是,它们上调的水平取决于我们实验中使用的压力因子的类型。对SvGls含量的分析表明,在所有应用条件下生长的HR都能够合成和积累几种SvGls,但在不同条件下样品之间的浓度不同。在所有测试条件下,HR中莱鲍迪甙A的含量均超过甜菊糖苷的含量。同时,仅在培养基中渗透压最低的培养液中证实了一些次要SvGls的存在,例如甜菊双糖苷和莱鲍迪甙F,而在添加了NaCl或PEG的培养基中培养的样品中也检测到莱鲍迪甙D。

重点
●在葡萄球菌的毛状根中合成了几种甜菊醇苷
●轻度或渗透性因素导致毛状根的氧化应激水平增强
●它与KAH表达水平的显着增加有关
●UGT的表达和甜菊糖苷的含量取决于培养条件。
更新日期:2020-06-22
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