糙皮侧耳锌指转录因子PoZF1的耐热性机理研究

In our country, the high temperature is one of the serious threats to the agricultural production of edible fungi. The molecular regulation mechanism of edible fungi under heat stress has not yet been clearly clarified. Transcription factors can resist heat stress by regulating the expression of specific functional genes, thus are considered key factors at the signal transduction terminal stage in response to heat stress. Previously, our research team found that after heat stress, expression level of Pleurotus ostreatus zinc finger transcription factor, PoZF1, increased 84 times. Homologous search showed PoZF1 to be an unknown protein. This study intends to use the strain, P. ostreatus CCEF89, as materials to construct the PoZF1 RNA interference mutants and over-expressing strains. Then we are going to study the PoZF1's effects on heat resistance, and its regulatory role on the main functional genes in response to heat stress. This study will lay a theoretical basis for the study of P. ostreatus signal transduction, gene regulation etc. in heat stress response. It will also provide the theoretical evidence for heat-tolerant strain identification and molecular marker assisted breeding.

在我国，高温严重威胁农业式食用菌生产。高温胁迫下食用菌分子调控机制至今尚未清晰。转录因子通过调控特定功能基因的表达来抵御高温胁迫，是响应高温胁迫的信号传导最终端的关键因素。本团队前期研究发现高温胁迫后糙皮侧耳锌指蛋白转录因子PoZF1表达量提高84倍，同源比对结果显示是未知蛋白。本研究拟以糙皮侧耳菌株CCEF89为材料，构建该PoZF1的RNA干扰突变体和过表达菌株，研究该转录因子对糙皮侧耳耐热性的影响，以及其对高温胁迫响应的主要功能基因的调控作用。本研究将为糙皮侧耳高温胁迫响应信号转导、基因调控等研究奠定理论基础，为耐高温菌种鉴定及分子辅助育种提供理论依据。

糙皮侧耳（Pleurotus ostreatus）是我国重要的栽培食用菌之一，其农业式栽培易受高温天气影响，热胁迫响应一直是研究的热点。转录因子通过调控特定功能基因的表达来抵御高温胁迫，是响应高温胁迫的信号传导最终端的关键因素。.本研究发现糙皮侧耳高温胁迫后一个锌指蛋白转录因子表达量提高84倍，同源比对结果显示是未知蛋白，我们命名为PoZCP26。本研究采用根癌农杆菌介导的遗传转化方法，构建了糙皮侧耳PoZF26 RNA干扰突变体和过表达菌株，分析了PoZCP26在糙皮侧耳菌丝的生长、耐热性，结果显示PoZCP26正向调节了菌丝的生长速度和热胁迫后的恢复生长能力，PoZCP26的过表达增强了菌丝在热胁迫中的耐受性。此外，本研究还发现PoZCP26在一定程度上正调控了菌丝在栽培基质内的长速，PoZCP26受到干扰后明显延迟了原基的形成并抑制了其分化，进而导致子实体畸形或者发育受阻。这些结果表明PoZCP26不仅响应热胁迫，调控糙皮侧耳菌丝热耐受性，而且调控子实体发育。转录组测序分析发现，PoZCP26可能通过调控G蛋白、疏水蛋白、DnaJ蛋白及参与蛋白泛素化和碳水化合物代谢相关酶的基因的表达来响应热胁迫。.本研究将为糙皮侧耳高温胁迫响应信号转导、子实体发育调控等研究奠定理论基础，为耐高温菌种鉴定及分子辅助育种提供理论依据。