MdERF2通过直接激活MdLACS1、MdCER1和MdCER6调控苹果采后表皮蜡质的形成

doi:10.1016/j.jia.2024.11.033

摘要/Abstract

摘要：

乙烯转录因子2 （ERF2）是植物生长，果实成熟、代谢和抵御胁迫所必需的。为了深入了解MdERF2在控制苹果表皮蜡质合成中的功能，本文利用MdERF2超表达（ERF2-OE）和沉默（ERF2-AN）载体，通过侵染苹果果实和/或愈伤组织，研究了蜡质合成有关基因表达，蜡质组成、含量和超微结构的变化，并利用凝胶电泳迁移实验（EMSAs）和双荧光素酶报告基因分析（DLRs）技术，鉴定了MdERF2直接调控的蜡质合成相关的基因。结果表明，EFR2-OE上调了MdLACS2，MdCER1，MdCER4和MdCER6等蜡质合成相关基因表达，而MdERF2沉默则抑制了上述基因的表达。同时，不同的MdERF2表达水平，还影响了蜡质的结构和积累。ERF2-OE处理显著增加了蜡质中烷烃和酮的比例，减少了脂肪酸和酯的比例。此外，EMSAs和DLRs实验还证明，MdERF可以与MdLACS2，MdCER1和MdCER6启动子部位的GCC-box元件直接结合，激活他们的转录水平。上述结果证明，MdERF2可以靶向上调MdLACS2，MdCER1和MdCER6基因的表达，从而改变了苹果表皮蜡质的组成、含量和微观结构。

Abstract:

Ethylene response factors 2 (ERF2) are essential for plant growth, fruit ripening, metabolism, and resistance to stress. In this study, the expression levels of the genes for MdERF2 implicated in the biosynthesis, composition and ultrastructure of fruit cuticular wax in apple (Malus domestica) were studied by the transfection of apple fruit and/or calli with MdERF2-overexpression (ERF2-OE) and MdERF2-interference (ERF2-AN) vectors. In addition, the direct target genes of MdERF2 related to wax biosynthesis were identified using electrophoretic mobility shift assays (EMSAs) and dual-luciferase reporter (DLR) assays. The findings indicated that the expression levels of four wax biosynthetic genes, long-chain acyl-CoA synthetase 2 (MdLACS2), eceriferum 1 (MdCER1), eceriferum 4 (MdCER4), and eceriferum 6 (MdCER6), were upregulated by ERF2-OE. In contrast, the expression levels of these genes were inhibited when MdERF2 was silenced. Furthermore, the overall structure and accumulation of fruit cuticular wax were influenced by the expression level of MdERF2. Treatment with ERF2-OE significantly increased the proportions of alkanes and ketones and reduced the proportions of fatty acids and esters. In addition, the EMSAs and DLR assays demonstrated that MdERF2 could bind directly to GCC-box elements in the promoters of MdLACS2, MdCER1, and MdCER6 to activate their transcription. These results confirmed that MdERF2 targets the up-regulation of expression of the MdLACS2, MdCER1, and MdCER6 genes, thereby altering the composition, content, and microstructure of apple epidermal wax.

Key words: apple fruit , wax , ethylene response factor 2 , target genes

Xinyue Zhang, Xinhua Zhang, Wenwen Sun, Meng Lü, Yefei Gu, Sarfaraz Hussain, Xiaoan Li, Maratab Ali, Fujun Li. MdERF2通过直接激活MdLACS1、MdCER1和MdCER6调控苹果采后表皮蜡质的形成[J]. Journal of Integrative Agriculture, 2025, 24(6): 2229-2239.

Xinyue Zhang, Xinhua Zhang, Wenwen Sun, Meng Lv, Yefei Gu, Sarfaraz Hussain, Xiaoan Li, Maratab Ali, Fujun Li. MdERF2 regulates cuticle wax formation by directly activating MdLACS2, MdCER1 and MdCER6 of apple fruit during postharvest[J]. Journal of Integrative Agriculture, 2025, 24(6): 2229-2239.

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