MlTT8 enhances anthocyanin biosynthesis in Magnolia liliiflora by interacting with proteins and directly binding to the MlCHS promoter

IF 4.2 2区农林科学 Q1 HORTICULTURE

Scientia Horticulturae Pub Date : 2025-08-01 DOI:10.1016/j.scienta.2025.114355

Jing-Jing Wang, Chao Wu, Meng-Yi Dai, Zhuo-Wei Wang, Feng Tian, Sen-Yu Zhu, Xiao-Jun Chen, Ya-Mei Shen, Jing-Jing Ma

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引用次数: 0

Abstract

The IIIf bHLH (basic/helix-loop-helix) subfamily controls plant anthocyanin production, but the specific regulatory mechanisms in Magnolia liliiflora are not well understood. Phylogenetic analysis of IIIf bHLH members identified anthocyanin pathway regulators, revealing two members in M. liliiflora. Among them, TRANSPARENT TESTA 8 (TT8) was identified as a potential positive regulator for further investigation based on sequence alignment. Nuclear localization was observed for MlTT8 and exhibited high expression levels in floral buds, particularly during the initial stages. MlTT8 overexpression enhanced anthocyanin accumulation and upregulated biosynthetic genes in Nicotiana tabacum. Yeast Two-hybrid (Y2H) analysis combined with bimolecular fluorescence complementation (BiFC) assays confirmed the formation of active MlTT8- MlMYB75 (R2R3-MYB transcription factor 75), MlTT8-MlTTG1 (Transparent Testa Glabra 1), MlTT8- MlMYC1 (MYELOCYTOMATOSIS 1) and MlTT8-MlTT8 protein complexes. Furthermore, it is demonstrated that MlTT8 directly activates the Chalcone Synthase (CHS) promoter to enhance anthocyanin synthesis through Yeast One-hybrid system (Y1H) combined with dual-Luciferase Reporter system (dual-Luc). These findings elucidated a regulatory module in which MlTT8 forms complex with other proteins and directly activates to enhance anthocyanin biosynthesis.

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MlTT8通过与蛋白相互作用和直接结合MlCHS启动子促进白玉兰花青素的生物合成

IIIf bHLH（碱性/螺旋-环-螺旋）亚家族控制植物花青素的产生，但在白玉兰中的具体调控机制尚不清楚。IIIf bHLH成员的系统发育分析鉴定了花青素途径调节因子，揭示了百合中两个成员。其中，TRANSPARENT TESTA 8 （TT8）被鉴定为潜在的正调控因子，可通过序列比对进一步研究。MlTT8的核定位在花芽中表现出高表达水平，特别是在花芽的初始阶段。MlTT8过表达增强了烟草花青素积累，上调了生物合成基因。酵母双杂交（Y2H）分析结合双分子荧光互补（BiFC）检测证实了活性MlTT8- MlMYB75 （R2R3-MYB转录因子75）、MlTT8- mlttg1 （Transparent Testa Glabra 1）、MlTT8- MlMYC1 （MYELOCYTOMATOSIS 1）和MlTT8-MlTT8蛋白复合物的形成。此外，MlTT8通过酵母单杂交系统（Y1H）与双荧光素酶报告子系统（dual-Luc）结合，直接激活查尔酮合成酶（Chalcone Synthase， CHS）启动子，促进花青素合成。这些发现阐明了MlTT8与其他蛋白形成复合物并直接激活以促进花青素生物合成的调控模块。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Scientia Horticulturae 农林科学-园艺

CiteScore

8.60

自引率

4.70%

发文量

796

审稿时长

47 days

期刊介绍： Scientia Horticulturae is an international journal publishing research related to horticultural crops. Articles in the journal deal with open or protected production of vegetables, fruits, edible fungi and ornamentals under temperate, subtropical and tropical conditions. Papers in related areas (biochemistry, micropropagation, soil science, plant breeding, plant physiology, phytopathology, etc.) are considered, if they contain information of direct significance to horticulture. Papers on the technical aspects of horticulture (engineering, crop processing, storage, transport etc.) are accepted for publication only if they relate directly to the living product. In the case of plantation crops, those yielding a product that may be used fresh (e.g. tropical vegetables, citrus, bananas, and other fruits) will be considered, while those papers describing the processing of the product (e.g. rubber, tobacco, and quinine) will not. The scope of the journal includes all horticultural crops but does not include speciality crops such as, medicinal crops or forestry crops, such as bamboo. Basic molecular studies without any direct application in horticulture will not be considered for this journal.