ELUCIDATION OF LIGNIFICATION MECHANISM OF WOODY PLANTS BY GENETIC ENGINEERING

基因工程阐明木本植物木质化机制

• 批准号: 07406008
• 负责人: MOROHOSHI Noriyuki
• 金额: $ 16.58万
• 依托单位: TOKYO UNIVERSITY OF AGRICULTURE AND TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07406008/
• 关键词: LIGNIN; LIGNIN BIOSYNTHETIC GENE; PHENYLALANINE AMMONIA-LYASE (PAL); CINNAMATE 4-HYDROXYLASE (CA4H); O-METHYLTRANSFERASE (OMT); PEROXIDASE (PO); TOBACCO; POPLAR; リグニン合成遺伝子; 免疫組織化学; 交雑ヤマナラシ; 木質化; 遺伝子; P.Kitakamiensis

To elucidate the lignification mechanism of woody plants from a viewpoint of molecular biology, some genes encoding phenylalanine ammonia-ly ase (PAL), cinnamate 4-hydroxylase (CA4H), O-methyltransferase (OMT) and peroxidase (PO) were isolated from the hybrid aspen (Populus kitakamiensis). Of these genes, PAL (palg2b), CH4H (cyp73a), OMT (homt1), and PO (prxA3a, prxA4a) were identified as the genes involved in lignification by the northem hybridization and the expression observation on tissue slices with reporter gene (GUS gene).These transformants of tobacco and poplar controlling the lignin biosynthesis by antisense RNA method with those identified genes were formed. The transformants concerning OMT have lower OMT activity and lignin content than those of normal tobacco and poplar, and have a characteristic lignin chemical structure consisting of lower amount of condensed units, and more of guaiacyl unit and degradable chemical structures.

为了从分子生物学的角度阐明木本植物的木质化机制，从杂交白杨（Populus kitakamiensis）中分离了一些编码苯丙氨酸解氨酶（PAL）、肉桂酸4-羟化酶（CA4H）、O-甲基转移酶（OMT）和过氧化物酶（PO）的基因。通过Northem杂交和报告基因（GUS基因）在组织切片上的表达观察，将这些基因中的PAL（palg2b）、CH4H（cyp73a）、OMT（homt1）和PO（prxA3a、prxA4a）确定为参与木质化的基因。这些烟草和杨树的转化子通过以下方式控制木质素生物合成： 利用这些已鉴定的基因形成反义RNA方法。 OMT转化体比正常烟草和杨树具有更低的OMT活性和木质素含量，并且具有由较少量的缩合单元、较多的愈创木基单元和可降解化学结构组成的特征性木质素化学结构。

项目成果

Y.Osakabe et al.: "Immunocytochemical localization of phenylalanine ammonia-lyase in tissues of Populus kitakamiensis." Planta. 200. 13-19 (1996)

Y.Osakabe 等人：“北上杨组织中苯丙氨酸解氨酶的免疫细胞化学定位。”

X.Zhan et al.: "A new approach based on the leaf disk method for Agrobacterlum medlated transformation and regeneration of aspen." Plant Science. 123. 105-112 (1997)

X.Zhan 等人：“一种基于叶盘法的农杆菌介导白杨转化和再生的新方法。”

T.Hayakawa,et al.: "Molucular cloning and tissue-specific expression on two genes that encode caffeic acid and O-methyltransgerases from Populus kitakamiensis." Plant Science. 113. 157-165 (1996)

T.Hayakawa 等人：“北上杨中编码咖啡酸和 O-甲基转移酶的两个基因的分子克隆和组织特异性表达。”

Y.Osakabe et al.: "Characterization of the structure and determination of mRNA levels of the phenylalanine ammonia-lyase gene family from Populus kitakamiensis." Plant Molecular Biology. 28. 1133-1141 (1995)

Y.Osakabe 等人：“北上杨苯丙氨酸解氨酶基因家族的结构表征和 mRNA 水平测定。”

Y.Osakabe et al.: "Molecular cloning and tissue-specific expression of two genes that encode caffeic acid and O-methyltransferases from Populus kitakamiensis." Plant. 200. 13-19 (1996)

Y.Osakabe 等人：“编码北上杨的咖啡酸和 O-甲基转移酶的两个基因的分子克隆和组织特异性表达。”