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Degradation of phenolic compounds by transgenic trees

转基因树对酚类化合物的降解

基本信息

批准号：
14380271
负责人：
KIMURA Tetsuya
金额：
$ 9.47万
依托单位：
Mie University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14380271/
关键词：
chlorocatechol rice poplar PHT1 Arabidopsis ファイトレメディエーション有機塩素系化合物 Relstonia

项目摘要

Utilization of transgenic trees as phytoremediation is considered to be one of the cost-effective ways for successive degradation of chlorinated aromatic compounds such as dioxins which are resistant to degradation for decades. Bacterial genes cbnA encoding chlorocatechol dioxigenase and cbnB encoding chloromuconate cycloisomerase from Ralstonia eutropha NH9, catalyzes one of the important steps for degradation of chlorocatechols and cleaves the aromatic ring of 3-chlorocatechol to produce toxically reduced 2-chloromuconate and muconolactone.In this study, we constructed a binary vector pCAMBIA-E7131-cbnA-cbnB transcribing cbnA and cbnB gene under the control of CaMV 35S promoter with enhancers, and introduced to rice and hybrid poplar (Populus tremula x tremuloides) by Agrobacterium mediated transformation. Integration of the gene to their chlomosomes was confirmed by PCR and the transgenic lines were isolated for subsequent analyses. Accumulation of cbnA and cbnB translation was visu … More alized by Western bloting and chlorocatechol dioxigenase activity was quantitatively detected by HPLC assay despite its high GC content (65%). When poplar transgenic calli was incubated for 2 hrs in the presence of 1 mM of 3-chlorocatechol, increase of a degradation product 2-chloromuconate peak signal was observed in parallel with reduced peak of 3-chlorocatechol substrate. The results showed that the cbnA gene and cbnB gene were successfully expressed in rice and poplar tree. These kinds of transgenic plants will be used for phytoremediation of environmental pollutants such as chlorinated aromatic compounds.For phytoremediation of pollutants in contaminated soil, the genes intorduced into plant cells should be expressed in root tissues since most of the contaminated chemicals are in soil. To express the genes in root tissues, strong promoters for root specific expression are necessary. To isolate these kinds of promoters, we selected the PHT1 promoter of Arabidopsis thaliana. GUS reporter analysis of PHT1 promoter in rice and Arabidopsis indicated that this promoter is useful for gene expression in root tissues both in monocot and dicot plants. Less

利用转基因树木进行植物修复被认为是一种具有成本效益的方法，可以连续降解几十年来难以降解的二恶英等氯代芳香族化合物。来自真养罗尔斯通氏菌（Ralstonia eutropha）NH 9的细菌基因cbnA和cbnB编码氯邻苯二酚二加氧酶和氯粘康酸环异构酶，催化氯邻苯二酚降解的重要步骤之一，并切割3-氯邻苯二酚的芳环以产生毒性还原的2-氯粘康酸和粘康酸内酯。我们构建了在带有增强子的CaMV 35 S启动子控制下转录cbnA和cbnB基因的双元载体pCAMBIA-E7131-cbnA-cbnB，并通过农杆菌介导的转化将其导入水稻和杂交白杨（Populus tremula x tremuloides）中。通过PCR证实基因整合到它们的染色体中，并分离转基因系用于随后的分析。cbnA和cbnB翻译的积累是可见的。 ...更多信息尽管其GC含量高（65%），但通过HPLC法定量检测氯儿茶酚二氧合酶活性。当白杨转基因愈伤组织在1 mM的3-氯儿茶酚存在下孵育2小时时，观察到降解产物2-氯粘康酸盐峰信号的增加与3-氯儿茶酚底物峰的降低平行。结果表明，cbnA基因和cbnB基因在水稻和白杨中得到了成功表达。这类转基因植物将用于氯代芳香族化合物等环境污染物的植物修复，而污染土壤中的污染物大多存在于土壤中，因此，在污染土壤的植物修复中，导入植物细胞的基因应在根组织中表达。为了在根组织中表达这些基因，需要用于根特异性表达的强启动子。为了分离这些类型的启动子，我们选择了拟南芥的PHT 1启动子。GUS报告基因分析表明，水稻和拟南芥中的PHT 1启动子在单子叶植物和双子叶植物的根组织中均能有效表达基因。少

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

プロモーターDNA断片及び遺伝子発現の制御方法

启动子DNA片段和控制基因表达的方法

DOI：
发表时间：
2003
期刊：
影响因子：
0
作者：
通讯作者：

Promoter of Arablidopsis thalliana phosphate transporter gene drives root-specific expression of transgene in rice

拟南芥磷酸转运蛋白基因的启动子驱动水稻根部特异性表达转基因

DOI：
发表时间：
2005
期刊：
Journal of Bioscience and Biotechnology 99
影响因子：
0
作者：
小山貴芳他10名
通讯作者：
小山貴芳他10名

Promoter of Arabidopsis thaliana phosphate transporter gene drives root specific expression of transgenic rice.

拟南芥磷酸转运蛋白基因的启动子驱动转基因水稻的根部特异性表达。

DOI：
发表时间：
2005
期刊：
J.Biosci.Bioeng. 99-1
影响因子：
0
作者：
T.Koyama;T.Ono;M.Shimizu;T.Jinbo;R.Mizuno;K.Tomita;N.Mitsukawa;T.Kawazu;T.Kimura;K.Ohmiya;K.Sakka
通讯作者：
K.Sakka

Biotechnology of lignocellulose degradation and biomass utilization