The interaction of MYB, bHLH and WD40 transcription factors in red pear (Pyrus pyrifolia) peel
The interaction of MYB, bHLH and WD40 transcription factors in red pear (Pyrus pyrifolia) peel
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DOI：
10.1007/s11103-021-01160-w
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发表时间：
2021-06-11
影响因子：
5.1
通讯作者：
Li, Kunzhi
中科院分区：
文献类型：
--
作者：
Cui, Daolei;Zhao, Shuxin;Li, Kunzhi
关键信息阳光增强果皮颜色，并显着上调PyMYB 10和PybHLH基因的表达。MYB-bHLH-WD 40转录复合物在光照条件下形成，参与调控果皮中花青素苷的积累。花青苷是云南红梨果皮中的主要色素。Nak.）。转录激活蛋白复合物，涉及MYB，bHLH和WD 40的转录因子类的成员，调节花青素苷的生物合成。在红梨果皮中检测到该复合物。为了阐明PyMYB 10、PybHLH和PyWD 40的相互作用，将果实套袋，然后在去除袋后0、3、5和7天收集果皮样品。将样品用于蛋白质印迹和蛋白质相互作用分析。结果表明，光照能显著提高果皮色泽，并显著上调PyMYB 10和PybHLH基因的表达。免疫共沉淀（Co-IP）分析表明PybHLH与PyMYB 10或PyWD 40相互作用，PyMYB 10与PyWD 40相互作用。使用洋葱细胞作为模型系统，双分子荧光互补（BiFC）证实了这些相互作用，并表明相互作用定位到细胞核。GST Pull down和Far-Western blotting分析表明PybHLH分别与PyMYB 10和PyWD 40相互作用，PyMYB 10与PyWD 40相互作用。此外，EMSA分析表明PyMYB 10可以直接结合到编码花青素生物合成酶花青素合成酶（PyANS）的基因的启动子上。综上所述，这些结果表明PyMYB 10、PybHLH和PyWD 40转录因子形成三元复合体调控云南红梨花色素苷的生物合成和积累。
Key message Sunlight enhanced peel color and significantly up-regulated the expression of PyMYB10 and PybHLH genes. MYB-bHLH-WD40 transcriptional complex forms in the light and is involved in regulating anthocyanin accumulation in the peel. Anthocyanin is the major pigment in the peel of Yunnan red pear (Pyrus pyrifolia (Burm.) Nak.). A transcriptional activation protein complex, involving members of the transcription factor classes of MYB, bHLH and WD40, regulates anthocyanin biosynthesis. This complex was examined in the peel of red pear. In order to clarify the interaction of PyMYB10, PybHLH and PyWD40, fruit were bagged then peel samples collected 0, 3, 5, and 7 days after bag removal. Samples were used for Western blotting and protein interaction analysis. The results showed that sunlight enhanced peel color and significantly up-regulated the expression of both PyMYB10 and PybHLH genes. Co-immunoprecipitation (Co-IP) analysis showed that PybHLH interacted with PyMYB10 or PyWD40, and PyMYB10 interacted with PyWD40. Using onion cells as a model system, bimolecular fluorescence complementation (BiFC) confirmed these interactions and showed that the interaction localized to the nuclei. GST Pull down and Far-Western blotting assays demonstrated that PybHLH interacted with PyMYB10 or PyWD40, respectively, and PyMYB10 interacted with PyWD40 in vitro. In addition, EMSA assay showed that PyMYB10 can directly bind to the promoter of the gene encoding the anthocyanin biosynthesis enzyme anthocyanidin synthase (PyANS). Taken together, these results showed that the ternary complex of PyMYB10, PybHLH and PyWD40 transcription factors forms to regulate anthocyanin biosynthesis and accumulation in Yunnan red pear.