Production mechanism of β-thujaplicin by cell cultures of woody species

木本植物细胞培养生产β-侧柏酚素的机制

基本信息

批准号：
13306013
负责人：
SAKAI Kokki
金额：
$ 25.63万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

β-Thujaplicin, called as hinokitiol in Japanese, is one of plant tropolones, contributing to the durability of heartwood of many Cupressaceae species due to its broad antifungal and antibacterial spectra. We have studied the mechanism of the production of β-thujaplicin by cell cultures of Cupressus lusitanica, a species in the Curpressaceae family.At first, a signal transduction pathway from elicitor receptor(s) on the cell surface to the onset of β-thujaplicin biosynthesis was investigated. It was elucidated by accelerator and/or inhibitor feeding experiments that G-protein, lipoxygenase, and protein kinases are involved in the signal transduction, and cAMP and Ca cation functioned as messengers. The jasmonate pathway is suggested to play an important role in the transduction system leading to β-thujaplicin production.Terpinolene synthase was found to be the most probable enzyme that is involved in biosynthesis of an intermediate monoterpene, most probably the direct precursor of β-thujaplicin. For cloning the gene of this enzyme by means of RT-PCR and cDNA screening, we obtained three DNA clones that may code the terpinolene synthase from a cDNA library of an elicitor-treated cell culture of C.lusitanica, because it was found that the enzyme was activated by the action of the elicitor. Then, the gene(s) that is involved in the β-thujaplicin biosynthesis was searched by the micro array method, for it was expected that mRNA(s) induce by the elicitation should reflect the β-thujaplicin biosynthesis gene(s). In cDNA library of C.lusitanica tells, it was observed that 48 clones exhibited significant differences in the strength between before and after the elicitor treatment. Function of these clones should be studied in future works.

β-thujaplicin在日语中被称为hinokitiol，是植物Troporones之一，由于其宽阔的抗真菌和抗菌光谱而导致许多杯状科物种的心脏耐用性。我们已经研究了通过cupressus lusitanica的细胞培养物（curpressaceae家族中的一种物种）的细胞培养物的生产机制。首先，研究了一种信号转导途径，从兴起受体（s）从细胞表面到β-硫非类生物合理的发作。它是通过加速剂和/或抑制剂喂养实验阐明的，即G蛋白，脂氧合酶和蛋白激酶参与信号传递，而cAMP和CA阳离子起着使者的作用。建议认为紫膜酸盐途径在导致β-噻帕环蛋白产生的转移系统中起重要作用。TERPINOLENE合酶是与中间单苯二酚的生物合成有关的最有问题的酶，很可能是β-thujaplicin的直接前体。为了通过RT-PCR和cDNA筛选克隆该酶的基因，我们获得了三个DNA克隆，可以从C. lusitanica c.lusitanica的cDNA库中编码Terpinolene合酶，因为发现该酶的细胞培养物的cDNA库，因为发现该酶被Elitoritor的作用激活了。然后，通过微阵列方法搜索参与β-胰蛋白酶生物合成的基因，因为预计会引起的mRNA影响应反映β-胰岛素生物合成基因。在C. lusitanica的cDNA文库中，观察到48个克隆在弹性剂治疗之前和之后的强度显着差异。这些克隆的功能应在未来的工作中进行研究。