Distribution of gibberellin-related antheridiogens in ferns and their mode of action

蕨类植物中赤霉素相关致瘤菌的分布及其作用方式

• 批准号: 04660134
• 负责人: YAMANE Hisakazu
• 金额: $ 1.34万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04660134/
• 关键词: Schizaeaceae; Lygodium flexuosum; Lygodium circinnatum; Mohria caffrorum; Ceratopteris richardii; antheridiogens; Lygodium japonicum; differential screening; Schizaeaceae; Lygodium japonicum; GA_<73>メチルエステル; Lygodium circinatum; GA_<73> methyl

Fern antheridiogens are divided into several groups based on cross-testing of biological activities. However, all the antheridiogens characterized so far were derived from Schizaeaceous ferns, and they are gibberellin-related compounds. The purpose of this study is to identify antheridiogens in three species of Scizaeaceous ferns (Lygodium flexuosum, L.circinnatum and Mohria caffrorum) and Ceratopteris richardii. The antheridiogens of C.richardii didn't show any cross-activity on Schizaeaceous ferns, but their biosynthesis was inhibited by treatment with gibberellin biosynthetic inhibitors. From culture media of prothallia of L.flexuosum and L.circinnatum, gibberellin A_<73> methyl ester (GA_<73>-Me) was identified by GC-MS as the principal antheridiogen. From theculture medium of prothallia of Mohria caffrorum, antheridic acid and 3-epi-GA_<63> were similarly identified as antheridiogens. The antheridiogens of C.richardii were shown to consist of at least two compounds, one of them being purified by HPLC to show a single peak on the chromatogrom.As a case study to investigate mode of action of the antheridiogens, cloning of GA_<73>-Me-inducing genes in L.japonicum prothallia was attempted. Differential screening on a cDNA library from GA_<73>-Me-treated prothallia was carried out using ^<32>P-labeled probes prepared from mRNA from GA_<73>-Me-treated and non-treated prothallia. However, no GA_<73>-Me-inducing gene has been obtained so far. We will attempt subtraction methods to clone antheridiogen-inducing genes.

根据生物学活性的交叉测试，蕨类植物的促精子剂被分为几类。然而，迄今为止所鉴定的所有促精子剂都来自于裂殖蕨，并且它们是赤霉素相关的化合物。对三种海金沙科蕨类植物（海金沙、卷纹海金沙和咖啡莫里亚）和水蕨中的精子器进行了鉴定。赤霉素生物合成抑制剂处理后，其生物合成受到抑制。用GC-MS法从弯叶百合和卷叶百合原叶体培养液中鉴定出赤霉素A<73>甲酯（GA_<73>-Me）为主要的雄激素。从咖啡Mohriacaffrorum原叶体培养液中，同样鉴定出了花药素和3-表-GA_2<63>为促雄剂。本研究以日本落叶松原叶体中GA_<73>-Me诱导基因的克隆为例，探讨了日本落叶松原叶体中GA_ -Me诱导基因的作用机制。以GA_<73>-Me处理的原叶体和未处理的原叶体<32>的mRNA为探针，对GA_<73>-Me处理的原叶体cDNA文库进行了差异筛选。但至今尚未获得GA_2<73>-Me诱导基因。我们将尝试用差减法克隆促精子生成素诱导基因。