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胞质溶胶参与茶叶中香叶基樱草糖苷的合成。

Cytosolic Is Involved in Geranyl -Primeveroside Production in Tea.

作者信息

Zhou Hanchen, Wang Shijie, Xie Hao-Fen, Liu Guofeng, Shamala Lubobi Ferdinand, Pang Jingyi, Zhang Zhengzhu, Ling Tie-Jun, Wei Shu

机构信息

State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, China.

Tea Research Institute, Anhui Academy of Agricultural Sciences, Huangshan, China.

出版信息

Front Plant Sci. 2022 May 11;13:833682. doi: 10.3389/fpls.2022.833682. eCollection 2022.

Abstract

Geraniol is a potent tea odorant and exists mainly as geranyl glycoside in . Understanding the mechanisms of geraniol biosynthesis at molecular levels in tea plants is of great importance for practical improvement of tea aroma. In this study, geraniol and its glycosides from tea plants were examined using liquid chromatography coupled with mass spectrometry. Two candidate geraniol synthase () genes () and two Nudix hydrolase genes ( and ) from the tea genome were functionally investigated through gene transcription manipulation and gene chemical product analyses. Our data showed that in tea leaves, levels of geranyl -primeveroside were dramatically higher than those of geranyl -glucoside, while free geraniol was undetectable in this study. A tempo-spatial variation of geranyl -primeveroside abundance in tea plants existed, with high levels in young and green tissues and low levels in mature or non-green tissues. Cytosolic CsNUDX1-cyto showed higher hydrolysis activity of geranyl-pyrophosphate to geranyl-monophosphate (GP) than did chloroplastidial CsNUDX1-chlo. A transgenic study revealed that expression of resulted in significantly more geranyl -primeveroside in transgenic compared with non-transgenic wild-type, whereas expression of had no effect. An antisense oligo-deoxynucleotide study confirmed that suppression of transcription in tea shoots led to a significant decrease in geranyl -primeveroside abundance. Additionally, transcript levels and geranyl -primeveroside abundances shared the same tempo-spatial patterns in different organs in the tea cultivar "Shucha Zao," indicating that is important for geranyl -primeveroside formation in tea plants. Results also suggested that neither of the two candidate GES genes in tea plants did not function as GES in transgenic . All our data indicated that is involved in geranyl -primeveroside production in tea plants. Our speculation about possible conversion from the chemical product of CsNUDX1-cyto to geranyl -primeveroside in plants was also discussed.

摘要

香叶醇是一种重要的茶叶香气成分,在茶叶中主要以香叶基糖苷的形式存在。从分子水平了解茶树香叶醇生物合成机制对于实际改善茶叶香气具有重要意义。本研究采用液相色谱-质谱联用技术对茶树中的香叶醇及其糖苷进行了检测。通过基因转录调控和基因化学产物分析,对茶树基因组中的两个候选香叶醇合酶(GES)基因(CsGES1和CsGES2)和两个Nudix水解酶基因(CsNUDX1和CsNUDX2)进行了功能研究。我们的数据表明,在茶叶中,香叶基樱草糖苷的含量显著高于香叶基葡萄糖苷,而在本研究中未检测到游离香叶醇。茶树中香叶基樱草糖苷含量存在时空变化,在幼嫩和绿色组织中含量较高,在成熟或非绿色组织中含量较低。胞质CsNUDX1-cyto对香叶基焦磷酸水解生成香叶基单磷酸(GP)的活性高于叶绿体CsNUDX1-chlo。转基因研究表明,与非转基因野生型相比,CsGES1基因在转基因烟草中表达导致香叶基樱草糖苷显著增加,而CsGES2基因表达则无影响。反义寡脱氧核苷酸研究证实,抑制茶树新梢中CsGES1基因的转录会导致香叶基樱草糖苷含量显著降低。此外,在茶树品种“舒茶早”的不同器官中,CsGES1转录水平和香叶基樱草糖苷含量具有相同的时空模式,表明CsGES1对茶树香叶基樱草糖苷的形成具有重要作用。结果还表明,茶树中的两个候选GES基因在转基因烟草中均未发挥GES的功能。我们所有的数据表明,CsNUDX1参与了茶树香叶基樱草糖苷的合成。我们还讨论了关于植物中CsNUDX1-cyto的化学产物可能转化为香叶基樱草糖苷的推测。

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