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Molecular and Functional Analysis of Pectins in Intercellular Attachment by T-DNA Tagging Using a Haploid Tobacco Tissue Culture System

使用单倍体烟草组织培养系统通过 T-DNA 标记对细胞间附着中的果胶进行分子和功能分析

基本信息

批准号：
14360204
负责人：
SATOH Shinobu
金额：
$ 8.45万
依托单位：
University of Tsukuba
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-14360204/
关键词：
tobacco pectin mutant borate arabinan callus 接着胚花粉花柱遺伝子グルクロン酸

项目摘要

Intercellular attachment is an essential process in the morphogenesis of multicellular organisms. A novel mutant, nolac-H18 (non-organogenic callus with loosely attached cells), generated by T -DNA transformation using leaf-disk-cultures of haploid Nicotiana plumbaginifolia, lost the ability to form tight intercellular attachments and adventitious shoots. The gene tagged with T -DNA, named NpGUT1 (glucuronyltransferase 1), was similar to the gene for the catalytic domains of animal glucuronyltransferases and was predominantly expressed in shoot and root apical meristems. The transformation of NpGUT1 complemented the nolac-H18 mutation and the expression of antisense NpGUT1 RNA produced crumbled shoots. The mutation caused defects in the glucuronic acid of rhamnogalacturonan II (RG-II) of pectin, which drastically reduced the formation of borate cross-linking of RG-II. NpGUT1, which encodes a novel glucuronyltransferase, is the first glycosyltransferase gene identified in pectin biosynthesis and is essential for intercellular attachment in plant meristems and tissues. In another mutant line nolac-H14, no neutral-sugar side chains, composed mainly of linear arabinan, were present. The gene tagged with T-DNA was cloned and named LARA1 (long arabinan related protein 1). The gene encodes a membrane protein and was predominantly expressed in shoot apex and root primordia. When nolac-H14 callus was cultured with arabinose, the phenotype of the mutant was recovered. These results suggest that LARA1 might be involved in the supply of UDP-arabinose to be putative arabinosyltransferase.

细胞间粘附是多细胞生物形态发生的重要过程。利用单倍体烟草（Nicotianaplumbaginifolia）的叶盘培养物通过T-DNA转化产生的新突变体nolac-H18（具有松散附着细胞的非器官发生愈伤组织）丧失了形成紧密细胞间附着和不定芽的能力。用T-DNA标记的基因，命名为NpGUT 1（葡萄糖醛酸转移酶1），与动物葡萄糖醛酸转移酶催化结构域的基因相似，并且主要在茎和根的顶端分生组织中表达。NpGUT 1的转化补充了nolac-H18突变，反义NpGUT 1 RNA的表达产生了破碎的芽。该突变引起的缺陷，在葡糖醛酸的鼠李糖半乳糖醛酸聚糖II（RG-II）的果胶，这大大减少了形成的硼酸盐交联的RG-II。NpGUT 1编码一种新的葡萄糖醛酸转移酶，是果胶生物合成中鉴定的第一个糖基转移酶基因，对植物分生组织和组织中的细胞间附着至关重要。在另一个突变株系nolac-H14中，不存在主要由线性阿拉伯聚糖组成的中性糖侧链。用T-DNA标记的基因被克隆并命名为LARA 1（long arabinan related protein 1）。该基因编码一种膜蛋白，主要在茎尖和根原基中表达。当nolac-H14愈伤组织与阿拉伯糖一起培养时，突变体的表型得到恢复。这些结果表明，LARA 1可能参与供应UDP-阿拉伯糖是假定的阿拉伯糖基转移酶。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

細胞接着とペクチン生合成

细胞粘附和果胶生物合成

DOI：
发表时间：
2002
期刊：
蛋白質・核酸・酵素 47
影响因子：
0
作者：
岩井宏暁;佐藤忍
通讯作者：
佐藤忍

Intercellular attachment and pectin biosynthesis

细胞间附着和果胶生物合成

DOI：
发表时间：
2002
期刊：
Protein, Nucleic Acid and Enzyme 47
影响因子：
0
作者：
H.Iwai;S.Satoh
通讯作者：
S.Satoh

A pectin glucuronyltransferase gene is essential for intercellular attachment in the plant meristem

DOI：
10.1073/pnas.252530499
发表时间：
2002-12-10
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Iwai, H;Masaoka, N;Satoh, S
通讯作者：
Satoh, S

Molecular and functional analysis of pectins in intercellular attachment with T-DNA tagging using the haploid tobacco tissue culture system.

使用单倍体烟草组织培养系统对细胞间附着的果胶进行 T-DNA 标记的分子和功能分析。