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

蕨类植物药剂根据生物活性的交叉测试被分成几组。然而，到目前为止，所有鉴定的雄激素都来自五味子科蕨类植物，它们都是赤霉素相关的化合物。本研究的目的是鉴定三种石杉科蕨类植物(卷叶蕨属Lygodium fleuosum、卷叶蕨属L.Circinnatum和咖啡色毛蕨属Mohria caffrorum)和山羊茅(Ceratopteris Richardii)中的花粉原。Richardii的雄蕊原在裂果蕨类植物上没有表现出任何交叉活性，但赤霉素生物合成抑制剂处理会抑制它们的生物合成。用气相色谱-质谱法(GC-MS)从柔嫩乳杆菌和卷曲乳杆菌的原生孢子培养基中鉴定出赤霉素A&lt；73&gt；甲酯(GA_&lt；73&gt；-Me)是主要的花粉诱导剂。从咖啡花原叶体的培养基中鉴定出花药酸和3-epi-GA_(63)&gt；通过高效液相色谱分离纯化了其中一种成分，并克隆了原花药中GA_&lt；73&Gt；-Me诱导基因，作为研究花药诱导剂作用方式的个案研究。利用从GA_lt；73&gt；Me处理和未处理的原叶中的mRNA制备的探针，对GA_&lt；73&gt；Me处理的原叶进行了差异筛选。然而，到目前为止，还没有获得GA_&lt；73&Gt；-me诱导基因。我们将尝试差减的方法来克隆去雄激素诱导基因。

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.