Zhao Qiuyu, Zhao Li, Shi Jiuzhi, Chen Xiaoxiao, Yang Zhenchao, Wu Yongjun
College of Life Sciences, Northwest A&F University, Yangling 712100, China.
College of Horticulture, Northwest A&F University, Yangling 712100, China.
Biomolecules. 2025 Jun 12;15(6):857. doi: 10.3390/biom15060857.
Lycopene, a natural pigment, is valuable for human health because of its strong antioxidant capacity. However, studies on the involvement of tomato miR394 in the regulation of lycopene have not been reported. The aim of this study was to reveal the molecular mechanism by which miR394 regulates lycopene synthesis by targeting ζ-carotene dehydrogenase (). The miR394-silenced transgenic tomato plants were constructed by short tandem target mimicry (STTM) technology, and the association between lycopene content and antioxidant capacity was analyzed by combining qRT-PCR, UV spectrophotometry, and a free radical scavenging assay. The targeting relationship between miR394 and was verified using a subcellular localization assay. The results showed that the silencing of miR394 significantly upregulated the expression of the ZDS gene and promoted lycopene accumulation. The antioxidant enzyme activities of STTM394 transgenic plants were significantly enhanced, and the free radical scavenging ability was obviously improved. Subcellular localization experiments confirmed that miR394 directly inhibited the chloroplast expression of . In conclusion, this study reveals for the first time that the miR394-ZDS module enhances the antioxidant capacity by regulating lycopene metabolism, which provides a new target for themolecular breeding of highly nutritious tomatoes.
番茄红素是一种天然色素,因其强大的抗氧化能力而对人体健康具有重要价值。然而,关于番茄miR394参与番茄红素调控的研究尚未见报道。本研究旨在揭示miR394通过靶向ζ-胡萝卜素脱氢酶(ZDS)调控番茄红素合成的分子机制。利用短串联靶标模拟(STTM)技术构建了miR394沉默的转基因番茄植株,并结合qRT-PCR、紫外分光光度法和自由基清除试验分析了番茄红素含量与抗氧化能力之间的关系。通过亚细胞定位试验验证了miR394与ZDS的靶向关系。结果表明,miR394的沉默显著上调了ZDS基因的表达并促进了番茄红素的积累。STTM394转基因植株的抗氧化酶活性显著增强,自由基清除能力明显提高。亚细胞定位实验证实miR394直接抑制ZDS在叶绿体中的表达。总之,本研究首次揭示了miR394-ZDS模块通过调控番茄红素代谢增强抗氧化能力,为高营养番茄的分子育种提供了新靶点。