Tanshinone production could be increased by the expression of SmWRKY2 in Salvia miltiorrhiza hairy roots
Tanshinone production could be increased by the expression of SmWRKY2 in Salvia miltiorrhiza hairy roots
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DOI：
10.1016/j.plantsci.2019.03.007
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发表时间：
2019-07-01
期刊：
影响因子：
5.2
通讯作者：
Kai, Guoyin
中科院分区：
文献类型：
--
作者：
Deng, Changping;Hao, Xiaolong;Kai, Guoyin
丹参酮是丹参中主要的生物活性二萜，广泛用于心脑血管疾病的治疗。然而，这些化合物的生物合成机制尚未得到充分解释。本研究分离了一个名为SmWRKY2的转录因子，并对其进行了功能表征。多序列分析表明该基因属于WRKY家族I亚群。表达模式显示SmWRKY2主要表达于茎和叶中，受茉莉酸甲酯（MeJA）诱导。亚细胞定位表明SmWRKY2定位于细胞核。在丹参毛状根中过表达SmWRKY2显著增加SmDXS2和SmCPS的表达，导致丹参酮积累增加，其中e -SmWRKY2-1系丹参酮总含量最高，为对照的1.83倍。同时，反义smwrky2系丹参酮产量略有下降。双荧光素酶实验表明，SmWRKY2可以正向调节SmDXS2和SmCPS的表达，但Y1H和EMSA实验表明，SmWRKY2仅结合SmCPS启动子的W-box。我们的研究表明SmWRKY2是丹参酮生物合成的正调控因子，主要通过激活SmCPS。该研究为SmWRKY2在丹参酮生物合成中的调控作用提供了新的思路。
Tanshinones are the main bioactive diterpenes in Salvia miltiorrhiza Bunge, are widely used for treating cardiovascular and cerebrovascular diseases. However, the biosynthetic mechanisms of these compounds have not yet been fully explained. In this study, a transcription factor named SmWRKY2 was isolated and functionally characterized. Multiple sequence analysis indicated it was classified into subgroup I of the WRKY family. Expression pattern showed that SmWRKY2 was mainly expressed in the stem and leaf and was inducible by methyl jasmonate (MeJA) treatment. Subcellular localization showed that SmWRKY2 was localized in the nucleus. Overexpression of SmWRKY2 in S. miltiorrhiza hairy roots significantly increased the expression of SmDXS2 and SmCPS, resulting in increased accumulation of tanshinones and the highest total tanshinone content was detected in OE-SmWRKY2-1 line, which was 1.83 times of the control. Meanwhile, tanshinone production was slightly reduced in the antisense-SmWRKY2 line. Dual-Luciferase assay showed that SmWRKY2 can positively regulate SmDXS2 and SmCPS expression, However, Y1H and EMSA experiments indicate that SmWRKY2 only binds to the W-box of the SmCPS promoter. Our study shows that SmWRKY2 is a positive regulator of tanshinone biosynthesis by mainly activating SmCPS. This study thus sheds new light on the regulatory role of SmWRKY2 in tanshinone biosynthesis.