三唑类杀菌剂是目前最重要的一类杀菌剂，而戊唑醇是其中用量最大的一种杀菌剂。因其广泛使用造成环境生物的潜在危害逐渐受到关注。前期研究发现戊唑醇对斑马鱼具有类雌激素样作用，对斑马鱼内分泌系统的HPG轴具有干扰效应，但这种干扰效应发生的关键环节及其分子机制等仍然不明。为此，本课题拟采用不同发育阶段的斑马鱼，结合高通量测定技术，研究鱼体内类固醇含量在戊唑曝露前后的变化，以及参与类固醇合成、转运、代谢的相关酶及其基因在药剂曝露前后表达量的变化，力图在类固醇/激素代谢途径上探究该药对斑马鱼类雌激素样干扰作用产生的关键环节和敏感期。进一步结合CRISPR/Cas9基因敲除技术，探讨类固醇生成因子SF-1在戊唑醇内分泌干扰中的作用，以期更好地解释戊唑醇对斑马鱼HPG轴干扰效应的分子通路机制。研究结果为揭示环境内分泌干扰的作用机制，探讨戊唑醇等三唑类农药的合理使用、环境保护和食品安全政策制定，提供理论依据。

Triazole fungicides are currently the most important class of fungicides, and the application amount of tebuconazole is the biggest one in all triazole fungicides in recent years. Due to its widely application, the potantial harm to environmental organisms is paid more and more attention. Previous study indicated that tebuconazole has the estrogen-like effect to zebrafish, which can disrupt the normal fuction of HPG axis of endocrine systerm. However, the key location where tebuconazole effects on and its molecular disrupting mechanism are still unknown. For this reason, combined with high throughput screening assay (such as UPLC-MS/MS and microarray screening of genes), developmental stages of zebrafish will be exposed to tebuconazole at environmental dosages to display the expression alternation of HPG axis related steroids and genes which participate in the synthesis, transportation and metabolism of these steroids and to discover the he key location and main lifestage where and when tebuconazole effects on. Furthermore, in order to better explain the molecular mechanism of tebuconazole disrupting effect on the HPG axis of zebrafish, steroidogenic factor 1 (SF-1) will be selected as target which tebuconazole effects on. The SF-1lack of phenotype mutant zebrafish by CRISPR/Cas9 knockdown assay will be cultured and used for exposure to tebuconazole and then the expression alternation of HPG axis related steroids and genes mentioned above will be also test. The prospective result will be benefit to explain the molecular mechanism of EEDs, provide the scientific evidence of reasonable application of tebuconzole and even triazole fungicides, policy making of environment protection and food safty, and so on.