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IbERF71 与 IbMYB340 和 IbbHLH2 一起,通过与紫色果肉甘薯(Ipomoea batatas L.)中的 IbANS1 启动子结合,共同调控花青素的积累。

IbERF71, with IbMYB340 and IbbHLH2, coregulates anthocyanin accumulation by binding to the IbANS1 promoter in purple-fleshed sweet potato (Ipomoea batatas L.).

机构信息

School of Food and Biological Engineering, Hefei University of Technology, Hefei, 230009, China.

Xuzhou Institute of Agricultural Sciences of the Xuhuai District of Jiangsu Province, Jiangsu Xuzhou Sweetpotato Research Center, Xuzhou, 221131, China.

出版信息

Plant Cell Rep. 2021 Jan;40(1):157-169. doi: 10.1007/s00299-020-02621-0. Epub 2020 Oct 21.

Abstract

The transcription factor (TF) IbERF71 forms a novel complex, IbERF71-IbMYB340-IbbHLH2, to coregulate anthocyanin biosynthesis by binding to the IbANS1 promoter in purple-fleshed sweet potatoes. Purple-fleshed sweet potato (Ipomoea batatas L.) is very popular because of its abundant anthocyanins, which are natural pigments with multiple physiological functions. TFs involved in regulating anthocyanin biosynthesis have been identified in many plants. However, the molecular mechanism of anthocyanin biosynthesis in purple-fleshed sweet potatoes has rarely been examined. In this study, TF IbERF71 and its partners were screened by bioinformatics and RT-qPCR analysis. The results showed that the expression levels of IbERF71 and partners IbMYB340 and IbbHLH2 were higher in purple-fleshed sweet potatoes than in other colors and that the expression levels positively correlated with anthocyanin contents. Moreover, transient expression assays showed that cotransformation of IbMYB340+IbbHLH2 resulted in anthocyanin accumulation in tobacco leaves and strawberry receptacles, and additional IbERF71 significantly increased visual aspects. Furthermore, the combination of the three TFs significantly increased the expression levels of FvANS and FvGST, which are involved in anthocyanin biosynthesis and transport of strawberry receptacles. The dual-luciferase reporter system verified that cotransformation of the three TFs enhanced the transcription activity of IbANS1. In addition, yeast two-hybrid and firefly luciferase complementation assays revealed that IbMYB340 interacted with IbbHLH2 and IbERF71 but IbERF71 could not interact with IbbHLH2 in vitro. In summary, our findings provide novel evidence that IbERF71 and IbMYB340-IbbHLH2 form the regulatory complex IbERF71-IbMYB340-IbbHLH2 that coregulates anthocyanin accumulation by binding to the IbANS1 promoter in purple-fleshed sweet potatoes. Thus, the present study provides a new regulatory network of anthocyanin biosynthesis and strong insight into the color development of purple-fleshed sweet potatoes.

摘要

转录因子 IbERF71 形成了一个新的复合物 IbERF71-IbMYB340-IbbHLH2,通过与紫色果肉甘薯中的 IbANS1 启动子结合,共同调控花青素生物合成。紫色果肉甘薯(Ipomoea batatas L.)因其富含花青素而非常受欢迎,花青素是具有多种生理功能的天然色素。在许多植物中已经鉴定出参与调控花青素生物合成的 TFs。然而,紫色果肉甘薯中花青素生物合成的分子机制很少被研究。在这项研究中,通过生物信息学和 RT-qPCR 分析筛选 TF IbERF71 及其伴侣。结果表明,与其他颜色相比,IbERF71 及其伴侣 IbMYB340 和 IbbHLH2 的表达水平在紫色果肉甘薯中更高,并且表达水平与花青素含量呈正相关。此外,瞬时表达试验表明,IbMYB340+IbbHLH2 的共转化导致烟草叶片和草莓接受器中花青素的积累,并且额外的 IbERF71 显著增加了视觉效果。此外,这三种 TF 的组合显著增加了参与草莓接受器花青素生物合成和运输的 FvANS 和 FvGST 的表达水平。双荧光素酶报告系统验证了三种 TF 的共转化增强了 IbANS1 的转录活性。此外,酵母双杂交和萤火虫荧光素酶互补测定表明,IbMYB340 与 IbbHLH2 和 IbERF71 相互作用,但 IbERF71 不能在体外与 IbbHLH2 相互作用。总之,我们的研究结果提供了新的证据表明 IbERF71 和 IbMYB340-IbbHLH2 形成了调控复合物 IbERF71-IbMYB340-IbbHLH2,通过与紫色果肉甘薯中的 IbANS1 启动子结合共同调控花青素的积累。因此,本研究为花青素生物合成的新调控网络提供了新的见解,并深入了解了紫色果肉甘薯的颜色发育。

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