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精细定位与候选基因分析影响辣椒果皮生物合成的 CaFCD1

Fine mapping and candidate gene analysis of CaFCD1 affecting cuticle biosynthesis in Capsicum annuum L.

机构信息

Engineering Research Center for Germplasm Innovation and New Varieties Breeding of Horticultural Crops, Key Laboratory for Vegetable Biology of Hunan Province, College of Horticulture, Hunan Agricultural University, Changsha, Hunan, China.

College of Horticulture, Nanjing Agricultural University, Nanjing, China.

出版信息

Theor Appl Genet. 2023 Mar 13;136(3):46. doi: 10.1007/s00122-023-04330-0.

Abstract

CaFCD1 gene regulates pepper cuticle biosynthesis. Pepper (Capsicum annuum L.) is an economically important vegetable crop that easily loses water after harvesting, which seriously affects the quality of its product. The cuticle is the lipid water-retaining layer on the outside of the fruit epidermis, which regulates the biological properties and reduces the rate of water-loss. However, the key genes involved in pepper fruit cuticle development are poorly understood. In this study, a pepper fruit cuticle development mutant fcd1 (fruit cuticle deficiency 1) was obtained by ethyl methanesulfonate mutagenesis. The mutant has great defects in fruit cuticle development, and the fruit water-loss rate of fcd1is significantly higher than that of the wild-type '8214' line. Genetic analysis suggested that the phenotype of the mutant fcd1 cuticle development defect was controlled by a recessive candidate gene CaFCD1 (Capsicum annuum fruit cuticle deficiency 1) on chromosome 12, which is mainly transcribed during fruit development. In fcd1, a base substitution within the CaFCD1 domain resulted in the premature termination of transcription, which affected cutin and wax biosynthesis in pepper fruit, as revealed by the GC-MS and RNA-seq analysis. Furthermore, the yeast one-hybrid and dual-luciferase reporter assays verified that the cutin synthesis protein CaCD2 was directly bound to the promoter of CaFCD1, suggesting that CaFCD1 may be a hub node in the cutin and wax biosynthetic regulatory network in pepper. This study provides a reference for candidate genes of cuticle synthesis and lays a foundation for breeding excellent pepper varieties.

摘要

CaFCD1 基因调控辣椒角质层生物合成。辣椒(Capsicum annuum L.)是一种经济上重要的蔬菜作物,收获后容易失水,严重影响其产品质量。角质层是果实表皮外的脂质保水层,调节生物特性并降低失水率。然而,参与辣椒果实角质层发育的关键基因知之甚少。本研究通过乙磺酸乙酯诱变获得了一个辣椒果实角质层发育突变体 fcd1(fruit cuticle deficiency 1)。该突变体在果实角质层发育方面存在严重缺陷,fcd1 的果实失水率明显高于野生型 '8214'系。遗传分析表明,突变体 fcd1 角质层发育缺陷的表型由位于第 12 号染色体上的隐性候选基因 CaFCD1(Capsicum annuum fruit cuticle deficiency 1)控制,该基因主要在果实发育过程中转录。在 fcd1 中,CaFCD1 结构域内的一个碱基替换导致转录提前终止,这影响了辣椒果实中的角质层和蜡质合成,GC-MS 和 RNA-seq 分析结果表明了这一点。此外,酵母单杂交和双荧光素酶报告基因检测验证了角质层合成蛋白 CaCD2 直接结合到 CaFCD1 的启动子上,表明 CaFCD1 可能是辣椒角质层和蜡质生物合成调控网络中的一个枢纽节点。本研究为角质层合成的候选基因提供了参考,为培育优良辣椒品种奠定了基础。

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