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转录因子IbNAC29正向调控甘薯中类胡萝卜素的积累。

The transcription factor IbNAC29 positively regulates the carotenoid accumulation in sweet potato.

作者信息

Xing Shihan, Li Ruijie, Zhao Haoqiang, Zhai Hong, He Shaozhen, Zhang Huan, Zhou Yuanyuan, Zhao Ning, Gao Shaopei, Liu Qingchang

机构信息

Key Laboratory of Sweet Potato Biology and Biotechnology, Ministry of Agriculture and Rural Affairs/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education, College of Agronomy & Biotechnology, China Agricultural University, Beijing 100193, China.

出版信息

Hortic Res. 2023 Feb 1;10(3):uhad010. doi: 10.1093/hr/uhad010. eCollection 2023 Mar.

Abstract

Carotenoid is a tetraterpene pigment beneficial for human health. Although the carotenoid biosynthesis pathway has been extensively studied in plants, relatively little is known about their regulation in sweet potato. Previously, we conducted the transcriptome database of differentially expressed genes between the sweet potato () cultivar 'Weiduoli' and its high-carotenoid mutant 'HVB-3'. In this study, we selected one of these candidate genes, , for subsequent analyses. IbNAC29 belongs to the plant-specific NAC (NAM, ATAF1/2, and CUC2) transcription factor family. Relative mRNA level in the HVB-3 storage roots was ~1.71-fold higher than Weiduoli. Additional experiments showed that the contents of α-carotene, lutein, β-carotene, zeaxanthin, and capsanthin are obviously increased in the storage roots of transgenic sweet potato plants overexpressing . Moreover, the levels of carotenoid biosynthesis genes in transgenic plants were also up-regulated. Nevertheless, yeast one-hybrid assays indicated that IbNAC29 could not directly bind to the promoters of these carotenoid biosynthesis genes. Furthermore, the level of was down-regulated, whose homologous genes in tomato can negatively regulate carotene accumulation. Yeast three-hybrid analysis revealed that the IbNAC29-IbMYB1R1-IbAITR5 could form a regulatory module. Yeast one-hybrid, electrophoretic mobility shift assay, quantitative PCR analysis of chromatin immunoprecipitation and dual-luciferase reporter assay showed that IbAITR5 directly binds to and inhibits the promoter activity of , up-regulating carotenoid biosynthesis gene . Taken together, is a potential candidate gene for the genetic improvement of nutritive value in sweet potato.

摘要

类胡萝卜素是一种对人体健康有益的四萜类色素。尽管类胡萝卜素生物合成途径在植物中已得到广泛研究,但关于其在甘薯中的调控机制却知之甚少。此前,我们构建了甘薯()品种‘维多利’与其高类胡萝卜素突变体‘HVB - 3’之间差异表达基因的转录组数据库。在本研究中,我们从这些候选基因中选择了一个基因进行后续分析。IbNAC29属于植物特有的NAC(NAM、ATAF1/2和CUC2)转录因子家族。HVB - 3贮藏根中的相对mRNA水平比维多利高约1.71倍。进一步的实验表明,在过量表达该基因的转基因甘薯植株的贮藏根中,α - 胡萝卜素、叶黄素、β - 胡萝卜素、玉米黄质和辣椒红素的含量显著增加。此外,转基因植株中类胡萝卜素生物合成基因的水平也上调。然而,酵母单杂交试验表明IbNAC29不能直接结合这些类胡萝卜素生物合成基因的启动子。此外,该基因的水平下调,其在番茄中的同源基因可负调控胡萝卜素积累。酵母三杂交分析表明IbNAC29 - IbMYB1R1 - IbAITR5可形成一个调控模块。酵母单杂交、电泳迁移率变动分析、染色质免疫沉淀定量PCR分析和双荧光素酶报告基因分析表明,IbAITR5直接结合并抑制该基因的启动子活性,上调类胡萝卜素生物合成基因。综上所述,该基因是甘薯营养价值遗传改良的一个潜在候选基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a162/10028406/5c27fb391fef/uhad010f1.jpg

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