Institute of Applied Mycology, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Key Laboratory of Agro-Microbial Resource Comprehensive Utilization, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, China.
Institute of Applied Mycology, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
Fungal Genet Biol. 2018 Sep;118:37-44. doi: 10.1016/j.fgb.2018.07.002. Epub 2018 Jul 10.
DnaJ proteins, termed heat shock proteins based on their molecular weight, function as molecular chaperones that play critical roles in regulating organism growth and development as well as adaptation to the environment. However, little has been reported on their gene function in higher basidiomycetes. Here, the heat shock protein 40 (LeDnaJ) gene was cloned and characterized from Lentinula edodes. RNA interference was used to explore the function of LeDnaJ in response to heat stress and Trichoderma atroviride. Integration of the target gene into the L. edodes genome was confirmed by Southern blot analysis, and the silence efficiency of LeDnaJ was analyzed by qRT-PCR. The results revealed that LeDnaJ silence caused defects in mycelial growth and resistance to heat stress and T. atroviride, but increased the mycelial density compared with the wild type (WT) strain S606. Additionally, the IAA content showed a more than 10-fold increase in the WT after heat stress, but an about two-fold increase in the two LeDnaJ RNAi transfortants (LeDnaJ-i-6 and LeDnaJ-i-8). Previous study has shown that enhanced IAA (indole-3-acetic acid) content enhanced the thermotolerance of the heat-sensitive strain YS3357. In this study, it was documented that IAA amendments could partly restore the resistance to T. atroviride and thermotolerance of the two LeDnaJ RNAi transformants. Overall, LeDnaJ is nvolved in fungal growth, T. atroviride resistance, and thermotolerance by regulating the IAA biosynthesis in L. edodes.
DnaJ 蛋白因其分子量而被称为热休克蛋白，作为分子伴侣发挥作用，在调节生物生长发育以及适应环境方面发挥着关键作用。然而，关于它们在高等担子菌中的基因功能报道甚少。本研究从香菇中克隆并鉴定了热休克蛋白 40（LeDnaJ）基因。通过 RNA 干扰技术研究了 LeDnaJ 在应对热应激和绿僵菌时的功能。通过 Southern blot 分析证实了目标基因整合到香菇基因组中，通过 qRT-PCR 分析了 LeDnaJ 的沉默效率。结果表明，LeDnaJ 沉默导致菌丝生长和耐热性及绿僵菌抗性缺陷，但与野生型（WT）菌株 S606 相比，菌丝密度增加。此外，WT 在热应激后 IAA（吲哚-3-乙酸）含量增加了 10 多倍，但两个 LeDnaJ RNAi 转化体（LeDnaJ-i-6 和 LeDnaJ-i-8）则增加了约两倍。先前的研究表明，增强的 IAA 含量增强了热敏感菌株 YS3357 的耐热性。本研究表明，IAA 添加剂可以部分恢复两个 LeDnaJ RNAi 转化体对绿僵菌的抗性和耐热性。总之，LeDnaJ 通过调节香菇中 IAA 的生物合成，参与真菌生长、绿僵菌抗性和耐热性。
