SmGRAS3 negatively responds to GA signaling while promotes tanshinones biosynthesis in Salvia miltiorrhiza
SmGRAS3 negatively responds to GA signaling while promotes tanshinones biosynthesis in Salvia miltiorrhiza
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DOI：
10.1016/j.indcrop.2019.112004
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发表时间：
2020-02-01
影响因子：
5.9
通讯作者：
Liang, Zongsuo
中科院分区：
文献类型：
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作者：
Li, Wenrui;Xing, Bingcong;Liang, Zongsuo
丹参是中药材丹参的原产地植物。由于丹参具有很好的药用价值，人们对高品质丹参的需求越来越大。丹参酮和丹酚酸是丹参的主要活性成分。赤霉素GA可诱导丹参酮和SAS的积累，但其机制尚不清楚。在这里，我们得到了一个GA响应蛋白SmGRAS3。SmGRAS3在丹参酮产生和积累的周皮中表达最高。SmGRAS3过表达和反义表达的结果表明，SmGRAS3是丹参酮生物合成的正调控因子。但过表达SmGRAS3抑制了SAS和GA的生物合成以及毛状根的生长，GA的应用可以部分恢复这些抑制作用。在转SmGRAS3基因的毛状根中，丹参酮、SAS和GA生物合成途径酶基因的表达发生了全面的变化。SmGRAS3是一种位于细胞核内的转录因子，具有转录激活活性。酵母单杂交(Y1H)和凝胶迁移率改变分析(EMSA)表明，SmGRAS3直接与KSL1启动子中的GARE基序结合并激活KSL1转录。我们的研究表明，SmGRAS3可能是丹参生物活性成分生物合成代谢工程的一个很好的潜在靶点。
Salvia miltiorrhiza Bunge is the origin plant of traditional Chinese medicine, Danshen. Demand of high-quality S. miltiorrhiza is increasing because of its excellent pharmaceutical value. Tanshinones and salvianolic acids (SAs) are the main active ingredients of S. miltiorrhiza. GA was reported to induce tanshinones and SAs accumulation, however, its mechanism was unclear. Here, we got a GA responsive protein, SmGRAS3. SmGRAS3 expressed highest in the periderm where tanshinones produced and accumulated. Results of SmGRAS3 over and antisense expression indicated that SmGRAS3 is a positive regulator of tanshinones biosynthesis in S. miltiorrhiza hairy roots. But overexpression of SmGRAS3 depressed SAs and GA biosynthesis as well as hairy roots growth, and application of GA could partially recover these depressions. Pathway enzyme genes expressions of tanshinones, SAs and GA biosynthesis were comprehensively changed in SmGRAS3 transgenic hairy roots. SmGRAS3 acted as a transcription factor that located in nucleus and has the transcription activation activity. Yeast one-hybrid (Y1H) and electrophoretic mobility shift assay (EMSA) showed that SmGRAS3 directly bound to GARE-motif in promoter of KSL1 and activated KSL1 transcription. Our study suggested that the SmGRAS3 may be a good potential target for further metabolic engineering of bioactive components biosynthesis in S. miltiorrhiza.