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基于图位克隆、功能互补和 RACE,鉴定了黄色果实番茄 2 编码的 PHYTOENE SYNTHASE 1 蛋白的跨剪接,通过该方法改变了果实颜色。

Identified trans-splicing of YELLOW-FRUITED TOMATO 2 encoding the PHYTOENE SYNTHASE 1 protein alters fruit color by map-based cloning, functional complementation and RACE.

机构信息

Department of Plant Science, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.

Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.

出版信息

Plant Mol Biol. 2019 Aug;100(6):647-658. doi: 10.1007/s11103-019-00886-y. Epub 2019 Jun 1.

Abstract

Found a trans-splicing of PHYTOENE SYNTHASE 1 alters tomato fruit color by map-based cloning, functional complementation and RACE providing an insight into fruit color development. Color is an important fruit quality trait and a major determinant of the economic value of tomato (Solanum lycopersicum). Fruit color inheritance in a yellow-fruited cherry tomato (cv. No. 22), named yellow-fruited tomato 2 (yft2), was shown to be controlled by a single recessive gene, YFT2. The YFT2 gene was mapped in a 95.7 kb region on chromosome 3, and the candidate gene, PHYTOENE SYNTHASE 1 (PSY1), was confirmed by functional complementation analysis. Constitutive over expression of PSY1 in yft2 increased the accumulation of carotenoids and resulted in a red fruit color, while no causal mutation was detected in the YFT2 allele of yft2, compared with red-fruited SL1995 cherry tomato or cultivated variety (cv. M82). Expression of YFT2 3' region in yft2 was significantly lower than in SL1995, and further studies revealed a difference in YFT2 post-transcriptional processing in yft2 compared with SL1995 and cv. M82, resulting in a longer YFT2 transcript. The alternatively trans-spliced allele of YFT2 in yft2 is predicted to encode a novel LT-YFT2 protein of 432 amino acid (AA) residues, compared to the 412 AA YFT2 protein of SL1995. The trans-spliced event also resulted in significantly down regulated expression of YFT2 in yft2 tomato, and the YFT2 allele suppressed expression of the downstream genes involved in the carotenoid biosynthesis pathway and carotenoids synthesis by a mechanism of the feed-forward regulation. In conclusion, we found that trans-splicing of YFT2 alters tomato fruit color, providing new insights into fruit color development.

摘要

通过图位克隆、功能互补和 RACE,发现 PHYTOENE SYNTHASE 1 的反式剪接改变了番茄果实颜色,为果实颜色发育提供了新的见解。颜色是果实的一个重要品质性状,也是番茄(Solanum lycopersicum)经济价值的主要决定因素。黄色果实樱桃番茄(品系 22)的果实颜色遗传由单个隐性基因 YFT2 控制。YFT2 基因定位于 3 号染色体上 95.7kb 区域,候选基因 PHYTOENE SYNTHASE 1(PSY1)通过功能互补分析得到确认。在 yft2 中组成型过表达 PSY1 增加了类胡萝卜素的积累,导致果实呈红色,而在 yft2 的 YFT2 等位基因中未检测到因果突变,与红色果实 SL1995 樱桃番茄或栽培品种(品系 M82)相比。与 SL1995 和品系 M82 相比,yft2 中 YFT2 3' 区域的表达明显较低,进一步研究表明,yft2 中 YFT2 转录后加工存在差异,导致 YFT2 转录本较长。与 SL1995 相比,yft2 中的 YFT2 替代反式剪接等位基因预测编码 432 个氨基酸(AA)残基的新型 LT-YFT2 蛋白,而 SL1995 的 YFT2 蛋白为 412AA。反式剪接事件还导致 yft2 番茄中 YFT2 的表达显著下调,YFT2 等位基因通过前馈调节机制抑制参与类胡萝卜素生物合成途径和类胡萝卜素合成的下游基因的表达。总之,我们发现 YFT2 的反式剪接改变了番茄果实颜色,为果实颜色发育提供了新的见解。

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