Data_Sheet_2_Label-Free Comparative Proteomics Analysis Revealed Heat Stress Responsive Mechanism in Hypsizygus marmoreus.xlsx

Heat stress is an important adverse environmental stress that influences the growth and development of Hypsizygus marmoreus (white var.). However, the molecular basis of heat stress response in H. marmoreus remains poorly understood. In this study, label-free comparative proteomic technique was applied to investigate global protein expression profile of H. marmoreus mycelia under heat stress. Confocal laser scanning microscope observation revealed that mycelia underwent autolysis and apoptosis under heat stress. Autolysis was mediated by upregulating the expression of cell wall degradation enzymes and inhibiting cell wall synthesis enzymes, and apoptosis might be induced by ROS and activation of caspases. TBARS analysis indicated that ROS was accumulated in H. marmoreus mycelia under heat stress. H. marmoreus induced antioxidant defense system by upregulating the expression of catalases, superoxide dismutases and peroxidases to prevent oxidative damage. MAPK cascade was found to be involved in heat stress signal transduction. The stress signal induced a ubiquitous defense response: inducible expression of different kinds of heat shock proteins. Trehalose synthesis enzymes were also upregulated, suggesting the accumulation of stress protector trehalose under heat stress. Besides, upregulated proteasome was identified, which could prevented the accumulation of non-functional misfolding proteins. To satisfy ATP depletion in heat response cellular processes, such as ROS scavenging, and protein folding and synthesis, enzymes involved in energy production (carbon metabolism and ATP synthesis) system were upregulated under heat stress. Taken together, these findings improve our understanding of the molecular mechanisms underlying the response of heat stress in H. marmoreus.

热应激作为一种重要的不利环境胁迫，对双孢蘑菇（白变种）的生长发育产生显著影响。然而，关于双孢蘑菇对热应激反应的分子基础，目前尚缺乏深入理解。本研究采用无标记比较蛋白质组学技术，探究了双孢蘑菇菌丝在热应激条件下的全局蛋白质表达谱。共聚焦激光扫描显微镜观察发现，菌丝在热应激下发生自溶和凋亡。自溶过程通过上调细胞壁降解酶的表达并抑制细胞壁合成酶的活性来实现，而凋亡可能由活性氧（ROS）的产生和caspase的激活所诱导。TBARS分析表明，在热应激条件下，双孢蘑菇菌丝中ROS的积累。双孢蘑菇通过上调过氧化氢酶、超氧化物歧化酶和过氧化物酶的表达，激活抗氧化防御系统，以防止氧化损伤。研究发现，丝裂原活化蛋白激酶（MAPK）级联反应参与热应激信号转导。应激信号诱导了普遍的防御反应：不同类型热休克蛋白的诱导表达。此外，海藻糖合成酶的表达也上调，这表明在热应激条件下，应激保护剂海藻糖的积累。此外，还鉴定出上调的蛋白酶体，它能够防止非功能性错误折叠蛋白的积累。为了满足热应激反应中细胞过程（如ROS清除、蛋白质折叠和合成）对ATP的消耗，参与能量产生（碳代谢和ATP合成）系统的酶在热应激条件下也被上调。综上所述，这些发现加深了我们对双孢蘑菇热应激反应分子机制的理解。