番茄 MADS-RIN 可调控 GAME5 的表达,促进果实中非苦味糖醛酸的生物合成。

IF 6.2 1区 生物学 Q1 PLANT SCIENCES The Plant Journal Pub Date : 2024-11-06 DOI:10.1111/tpj.17125
Yinhuan Xie, Yaping Xu, Huimin Jia, Ke Wang, Siyu Chen, Ting Ma, Yuanwei Deng, Zhaobo Lang, Qingfeng Niu
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引用次数: 0

摘要

众所周知,与防御相关的类固醇糖醛酸(SGA)代谢转变可消除成熟番茄果实的苦味和毒性。本研究旨在阐明 MADS-RIN(RIN)及其辅助因子对番茄果实中 SGA 代谢的影响。利用基于 CRISPR/Cas9 的基因编辑系统,我们突变了 RIN 和两个辅助因子基因(FUL1 和 FUL2)。在突变体中观察到的果实颜色和大小的变化反映了 RIN、FUL1 和 FUL2 对果实成熟的重叠和不同影响。根据 UPLC-MS/MS 分析,RIN 和辅助因子突变体中相对无毒的代谢物 esculeoside A 水平降低,但它们积累了有毒的 SGA 途径中间产物,这表明 RIN 及其辅助因子直接参与了 esculeoside A 的生物合成。转录组和 qPCR 分析检测到 GAME5 的表达下调,GAME5 编码介导 esculeoside A 生物合成的关键酶。ChIP-seq 和 ChIP-qPCR 分析证实 GAME5 是 RIN 的靶标。据观察,RIN 通过与 GAME5 启动子中的两个非经典 CArG-box 结合来激活 GAME5 的转录。此外,RIN 促进 SGA 的代谢与乙烯无关。总之,这些发现加深了我们对番茄果实成熟和 SGA 生物合成分子机制的理解。此外,这些发现可能有助于提高番茄果实的质量和安全性。
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Tomato MADS-RIN regulates GAME5 expression to promote non-bitter glycoalkaloid biosynthesis in fruit.

A well-known defense-associated steroidal glycoalkaloid (SGA) metabolic shift eliminates the bitterness and toxicity of ripe tomato fruits. This study was conducted to clarify the effects of MADS-RIN (RIN) and its cofactors on SGA metabolism in tomato fruits. Using a CRISPR/Cas9-based gene-editing system, we mutated RIN and two cofactor genes (FUL1 and FUL2). The observed changes to fruit color and size in the mutants reflected the overlapping and distinct effects of RIN, FUL1, and FUL2 on fruit ripening. According to a UPLC-MS/MS analysis, the RIN and cofactor mutants had decreased levels of the relatively non-toxic metabolite esculeoside A, but they accumulated toxic SGA pathway intermediates, suggesting RIN and its cofactors are directly involved in esculeoside A biosynthesis. Transcriptome and qPCR analyses detected the downregulated expression of GAME5, which encodes a key enzyme mediating esculeoside A biosynthesis. ChIP-seq and ChIP-qPCR analyses confirmed GAME5 is targeted by RIN. RIN was observed to activate GAME5 transcription by binding to two non-canonical CArG-boxes in the GAME5 promoter. Additionally, RIN promotes SGA metabolism independently of ethylene. Collectively, these findings enhance our understanding of the molecular mechanism governing tomato fruit ripening and SGA biosynthesis. Furthermore, they may be useful for improving tomato fruit quality and safety.

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来源期刊
The Plant Journal
The Plant Journal 生物-植物科学
CiteScore
13.10
自引率
4.20%
发文量
415
审稿时长
2.3 months
期刊介绍: Publishing the best original research papers in all key areas of modern plant biology from the world"s leading laboratories, The Plant Journal provides a dynamic forum for this ever growing international research community. Plant science research is now at the forefront of research in the biological sciences, with breakthroughs in our understanding of fundamental processes in plants matching those in other organisms. The impact of molecular genetics and the availability of model and crop species can be seen in all aspects of plant biology. For publication in The Plant Journal the research must provide a highly significant new contribution to our understanding of plants and be of general interest to the plant science community.
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