Functional characterisation of genes putatively involved in fruiting body formation of the cultivated edible mushroom Agrocybe aegerita

栽培食用菌茶树菇子实体形成相关基因的功能表征

• 批准号: 437330589
• 负责人: Dr. Florian Hennicke
• 金额: --
• 依托单位: Institut für Pflanzenschutz in Gartenbau und urbanem Grün
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/437330589?language=en
• 关键词: Functional; characterisation; genes; putatively; involved

On the molecular level, fruiting body formation by edible mushrooms is poorly understood. As a basis of fruiting body formation in basidiomycetes, sexual development is well studied in the split gill mushroom Schizophyllum commune and the ink cap Coprinopsis cinerea. In contrast to that, edible mushrooms are up to now less accessible with regard to functional genetics approaches. Agrocybe aegerita is a commercially cultivated edible mushroom which exhibits the rare feature of monokaryotic fruiting sensu stricto which means that it is able to fruit without any sexual mating beforehand. A recently published work exploring molecular tools for transformation and constitutive gene expression in A. aegerita now permits functional genetics-based approaches in this mushroom. Within the proposed project, up to now uncharacterised fruiting-related genes (FRGs) will be functionally characterised, starting with four genes, the differential expression of which during fruiting has already been validated and which are predicted to likely encode transcription factors. In addition to the already set up functional-genetics tools for constitutive gene of interest expression (‘overexpression’) from preliminary work, CRISPR/Cas9-based gene deletion is to be adapted for A. aegerita. Ultimately, commonalities and differences between exceptional monokaryotic and regular dikaryotic fruiting of A. aegerita will be defined. To achieve this, RNA-sequencing is to be conducted, employing wild type monokaryotic fruiting stages versus different monokaryotic fruiting phenotypes exhibited by two selected mutants generated within this project. This approach will also help to understand the mutagenised genes’ roles within the complex genetic network of predicted FRGs in A. aegerita. Using this model system, the expected results will further bridge gaps in the knowledge on molecular biology of fruiting body formation and will also provide first insights on what differentiates monokaryotic fruiting from regular dikaryotic fruiting in mushrooms.

在分子水平上，人们对食用菌子实体的形成知之甚少。作为担子菌子实体形成的基础，性发育在裂鳃菇Schizophyllum commune和墨盖Coprinopsis cinerea中得到了很好的研究。相比之下，到目前为止，食用蘑菇在功能遗传学方法方面还不太容易获得。绿草结菇是一种商业化栽培的食用菌，它具有罕见的单核严格结果感，这意味着它可以在没有任何性交配的情况下结果。最近发表的一项研究工作探索了A. aegerita转化和构成基因表达的分子工具，现在允许在这种蘑菇中使用基于功能遗传学的方法。在拟议的项目中，到目前为止尚未表征的结果相关基因（FRGs）将从四个基因开始进行功能表征，这些基因在结果期间的差异表达已经得到验证，并且被预测可能编码转录因子。除了从前期工作中已经建立的对感兴趣的组成基因表达（“过表达”）的功能遗传学工具外，基于CRISPR/ cas9的基因缺失也将适用于A. aegerita。最后，共同之处和差异之间的特殊单核和常规的双核结果将被定义。为了实现这一点，将进行rna测序，将野生型单核结果期与本项目中产生的两个选定突变体所表现出的不同单核结果表型进行比较。这种方法也将有助于理解突变基因在拟南毛线虫FRGs复杂遗传网络中的作用。利用该模型系统，预期的结果将进一步弥合子实体形成的分子生物学知识的空白，并将首次了解蘑菇单核子实体与常规双核子实体的区别。