高温严重影响香菇菌丝生长，甚至导致菌丝凋亡，但香菇不同菌株的耐热能力明显不同。申请者在前期研究中发现，香菇耐热菌株S606在40℃胁迫24 h之后，邻氨基苯甲酸合酶蛋白表达水平显著上升，但转录水平却明显下降，尤其是在培养基中添加邻氨基苯甲酸能显著提高菌丝耐热能力，但邻氨基苯甲酸合酶影响香菇耐热性的机制却并不清楚。本项目拟一方面采用基因过表达和基因沉默方法鉴定trpE的基因功能，应用免疫共沉淀技术筛选与TrpE互作的蛋白，并开展蛋白间的体内体外互作验证，明确TrpE与互作蛋白共同影响香菇耐热性的机制；另一方面采用组学方法，分析与TrpE相关的色氨酸合成代谢途径中酶基因及代谢产物的表达水平，对与耐热性相关的酶基因进行功能分析，揭示色氨酸合成代谢途径影响香菇耐热性的机制。本项目对深刻揭示真菌耐热机制和遗传基础，培育食用菌耐热品种，具有重要的科学意义和实践价值。

Heat stress often infects seriously the mycelia growth, even causes mycelia apoptosis of Lentinula edodes. However, various strains generally show differences in the thermos-tolerance. In our previous research, the protein level of anthranilate synthase (TrpE) was significantly up-regulated in thermo-tolerant strain S606 after heat treatment at 40℃ for 24 h whereas there was a down-regulated transcriptional level. Moreover, the thermo-tolerance of mycelia would be significantly enhanced when the anthranilate is added into medium with strain S606. However, the mechanism of this protein TrpE involved in heat tolerance in L. edodes has not been determined until until now. In this proposal, the function of trpE would be identified via gene over-expression and gene silencing, and the target proteins that were interacted with the TrpE would be baited out by Co-Immunoprecipitation. Putative protein-interactions would be validated via in vitro and in vivo for explicating the interaction mechanism during the thermo-tolerance process. On the other hand, the candidate gene expression levels and metabolite dynamics engaged in tryptophan synthesis metabolic pathways would be evaluated, and the function of heat-tolerance related genes would be identified to reveal the heat-tolerance mechanism related to tryptophan biosynthesis metabolic pathways in L. edodes. This proposal have great significance in theoretical and applied prospect on the heat-tolerance molecular mechanism, genetic basis and breeding thermo-tolerant varieties of the edible mushroom in the future.