Control of plant phenylpropanoid metabolism by heat sensitive promoter

热敏启动子对植物苯丙素代谢的控制

• 批准号: 12839001
• 负责人: YAMAKAWA Takashi
• 金额: $ 2.37万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12839001/
• 关键词: heat sensitive promoter; heat shock promoter; Nicotiana plumbagnifolia; hairy root; Nicotiaca plumbaginifolia; 二次代謝産物

The aim of this research is to control plant secondary metabolism by using heat-shock inducible promoter which is connected to the inserted gene in plant genome. Nicotiana plumbaginifolia was used as a model plant and chilli pepper(Capsicum frutescens) was used for the example of a crop. Chimera genes of Arabidopsis heat-shock protein(HSP)18.2 promoter and β-glucronidase(GUS) gene from E. coli or parsley phenylalanineammonia-lyase(PAL) gene were constructed and they were introduced into the hairy roots of N. plumbaginifolia. Their inducibility by heat shock treatment was investigated. GUS activity was induced by heat shock treatment at 40℃ for 2 hours on hairy roots and this inducibility was stable for 3-5 months. The inducibility was reproduced on 6 days after first heat shock treatment. However, GUS activity showed no change by heat shock treatment at 38-44℃ for 2 hours in hairy roots of chilli pepper. A parsley PAL activity also showed no change after heat shock treatment on hairy roots of N. plumbaginifolia, and sometimes the PAL activity decreased temporaly. These phenomena indicated that induced foreign PAL gene caused a co-supression. Intended increase of PAL activity was not observbed in this experiment, but it can be said that metabolism flux change using co-supression is possible by heat treatment of plant hairy root culture.

本研究的目的是利用与植物基因组中插入的基因相连接的热激诱导启动子来调控植物的次生代谢。以白叶烟草为模式植物，以辣椒(Capsicum Fuctescens)为作物实例。构建了拟南芥热休克蛋白(HSP18.2)启动子和大肠杆菌β-葡萄糖苷酶(GUS)基因或欧芹苯丙氨酸解氨酶(PAL)基因的嵌合体基因，并将其导入白花菜毛状根中。研究了它们在热休克处理中的诱导性。毛状根经40℃、2小时热休克处理后，GUS活性在3~5个月内稳定。在第一次热休克处理后第6天复制诱导性。而38-44℃的热激处理2小时，辣椒毛状根的GUS活性无明显变化。白花菜毛状根经热激处理后，其PAL活性也无明显变化，有时PAL活性呈暂时性下降。这些现象表明，外源PAL基因的诱导引起了共抑制。在本实验中没有观察到PAL活性的有意提高，但可以说，通过对植物毛状根培养物进行热处理，可以利用共抑制来改变代谢通量。