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

根据交叉测试的生物活性，蕨类花药属植物可分为几类。然而，迄今为止所鉴定的所有花药原均来自裂科蕨类植物，它们是赤霉素相关化合物。本研究的目的是鉴定三种剑科蕨类植物（Lygodium flexuosum， L.circinnatum和morhria caffrorum）和richardii角翅蕨的花药原。赤霉素对裂科蕨类植物无交叉作用，但其合成受到赤霉素生物合成抑制剂的抑制。用气相色谱-质谱联用技术从弯草和卷草原菌培养基中鉴定出赤霉素A_<73>-Me甲酯（GA_<73>-Me）为主要的抗氧化剂。从黑毛菌原菌培养基中，蒽醌酸和3-epi-GA_<63>同样被鉴定为蒽醌。richardii的花药原至少由两种化合物组成，其中一种化合物经HPLC纯化后在色谱上显示单峰。为了研究花药原的作用方式，本研究尝试克隆了原叶橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐橐。利用GA_<73>- me处理和未处理的原草体mRNA制备的^<32> p标记探针，对GA_<73>- me处理的原草体cDNA文库进行差异筛选。然而，目前尚未获得GA_<73>- me诱导基因。我们将尝试减法克隆花药诱导基因。

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.