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柑橘 UDP-糖基转移酶家族的全基因组分析及编码黄酮类化合物 1-2 鼠李糖基转移酶的 UGT 基因的特征

Genome-wide analysis of UDP-glycosyltransferase family in Citrus sinensis and characterization of a UGT gene encoding flavonoid 1-2 rhamnosyltransferase.

机构信息

Shandong Peanut Research Institute, Qingdao, Shandong 266100, PR China; Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China.

Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China.

出版信息

Int J Biol Macromol. 2024 Nov;280(Pt 2):135752. doi: 10.1016/j.ijbiomac.2024.135752. Epub 2024 Sep 17.

DOI:10.1016/j.ijbiomac.2024.135752
PMID:39299422
Abstract

UDP-glycosyltransferases (UGTs) play a crucial role in the glycosylation of secondary metabolites in plants, which is of significant importance for growth and response to biotic or abiotic stress. Despite the wide identification of UGT family members in various species, limited information is available regarding this family in citrus. In this study, we identified 87 UGT genes from the Citrus sinensis genome and classified them into 14 groups. We characterized their gene structures and motif compositions, providing insights into the molecular basis underlying discrepant functions of UGT genes within each evolutionary branch. Tandem duplication events were found to be the main driving force behind UGT gene expansion. Additionally, we identified numerous cis-acting elements in the promoter region of UGT genes, including those responsive to light, growth factors, phytohormones, and stress conditions. Notably, light-responsive elements were found with a frequency of 100 %. We elucidated the expression pattern of UGTs during fruit development in Citrus aurantium using RNA-seq and quantitative real-time PCR (qRT-PCR), revealing that 10 key UGT genes are closely associated with biosynthesis of bitter flavanone neohesperidosides (FNHs). Furthermore, we identified Ca1,2RhaT as a flavonoid 1-2 rhamnosyltransferase (1,2RhaT) involved in FNHs biosynthesis for the first time. Isolation and functional characterization of the gene Ca1,2RhaT from Citrus aurantium in vitro and in vivo indicated that Ca1,2RhaT encoded a citrus 1,2RhaT and possessed rhamnosyl transfer activities. This work provides comprehensive information on the UGT family while offering new insights into understanding molecular mechanisms regulating specific accumulation patterns of FNHs or non-bitter flavanone rutinosides (FRTs) in citrus.

摘要

UDP-糖基转移酶(UGTs)在植物次生代谢物的糖基化过程中发挥着关键作用,这对于植物的生长和应对生物或非生物胁迫至关重要。尽管在各种物种中广泛鉴定了 UGT 家族成员,但有关柑橘中的该家族的信息有限。在这项研究中,我们从柑橘基因组中鉴定了 87 个 UGT 基因,并将它们分为 14 组。我们对它们的基因结构和基序组成进行了特征描述,为不同进化分支中 UGT 基因功能差异的分子基础提供了见解。串联重复事件被发现是 UGT 基因扩张的主要驱动力。此外,我们还在 UGT 基因启动子区域鉴定了许多顺式作用元件,包括对光、生长因子、植物激素和胁迫条件响应的元件。值得注意的是,光响应元件的频率为 100%。我们使用 RNA-seq 和定量实时 PCR(qRT-PCR)研究了柑橘果实发育过程中 UGT 的表达模式,结果表明 10 个关键 UGT 基因与苦味黄烷酮新橙皮苷(FNHs)的生物合成密切相关。此外,我们首次鉴定了 Ca1,2RhaT 是参与 FNHs 生物合成的类黄酮 1-2 鼠李糖苷基转移酶(1,2RhaT)。从柑橘中分离和功能表征 Ca1,2RhaT 基因的体外和体内实验表明,Ca1,2RhaT 编码一个柑橘类 1,2RhaT,并具有鼠李糖基转移酶活性。这项工作提供了 UGT 家族的全面信息,同时为理解调控 FNHs 或非苦味黄烷酮鼠李糖苷(FRTs)在柑橘中特异性积累模式的分子机制提供了新的见解。

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