Cloning and functional characterization of a p-coumaroyl quinate/shikimate 3′-hydroxylase from potato (Solanum tuberosum)
Cloning and functional characterization of a p-coumaroyl quinate/shikimate 3′-hydroxylase from potato (Solanum tuberosum)
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DOI：
10.1016/j.bbrc.2018.01.075
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发表时间：
2018-02-05
影响因子：
3.1
通讯作者：
Tian, Li
中科院分区：
文献类型：
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作者：
Knollenberg, Benjamin J.;Liu, Jingjing;Tian, Li
绿原酸（Chlorogenic add，CGA）在保护植物免受病原菌侵害和促进人类健康方面具有重要作用.尽管CGA在马铃薯块茎中积累到高水平，但CGA生物合成的关键酶对香豆酰奎尼酸/希尔西马酸3 '-羟化酶（C3' H）尚未在马铃薯中分离和功能表征。在这项工作中，我们从马铃薯中克隆了StC 3 'H，并表明它分别催化对香豆酰莽草酸和对香豆酰奎尼酸形成咖啡酰莽草酸和CGA（咖啡酰奎尼酸），但在体外酶测定中对对香豆酸无活性。当StC 3 'H蛋白的表达被阻断，通过反义（AS）抑制块茎特异性patatin启动子的控制下，适度的变化，块茎产量以及在核心块茎组织中的酚类代谢产物观察到几个AS线。另一方面，AS和对照马铃薯品系对细菌病原体胡萝卜果胶杆菌表现出相似的反应。这些结果表明StC 3 ′ H参与马铃薯中的酚类代谢。他们还表明，马铃薯块茎核心组织中的CGA积累是一个复杂控制的过程，另外的C3 'H活性也可能参与马铃薯中的CGA生物合成。(C)2018爱思唯尔公司All rights reserved.
Chlorogenic add (CGA) plays an important role in protecting plants against pathogens and promoting human health. Although CGA accumulates to high levels in potato tubers, the key enzyme p-coumaroyl quinate/shilcimate 3'-hydroxylase (C3'H) for CGA biosynthesis has not been isolated and functionally characterized in potato. In this work, we cloned StC3'H from potato and showed that it catalyzed the formation of caffeoylshikimate and CGA (caffeoylquinate) from p-coumaroyl shikimate and p-coumaroyl quinate, respectively, but was inactive towards p-coumaric acid in in vitro enzyme assays. When the expression of StC3'H proteins was blocked through antisense (AS) inhibition under the control of a tuber specific patatin promoter, moderate changes in tuber yield as well as phenolic metabolites in the core tuber tissue were observed for several AS lines. On the other hand, the AS and control potato lines exhibited similar responses to a bacterial pathogen Pectobacterium carotovorum. These results suggest that StC3'H is implicated in phenolic metabolism in potato. They also suggest that CGA accumulation in the core tissue of potato tubers is an intricately controlled process and that additional C3'H activity may also be involved in CGA biosynthesis in potato. (C) 2018 Elsevier Inc. All rights reserved.