Data_Sheet_1_Differential Expression Characterisation of the Heat Shock Proteins DnaJB6, DnaJshv, DnaJB13, and DnaJB14 in Apis cerana cerana Under Various Stress Conditions.zip

As key pollinators, bees are frequently exposed to multiple environmental stresses and have developed crucial mechanisms by which they adapt to these stressors. However, the molecular bases mediated at the gene level remain to be discovered. Here, we found four heat shock protein DnaJB subfamily genes, DnaJB6, DnaJshv, DnaJB13, and DnaJB14, from Apis cerana cerana, that all have J domains in their protein sequences. The expression levels of DnaJB6 and DnaJshv were upregulated by different degrees of heat stress, and the transcript level of DnaJB14 was gradually upregulated as the degree of heat stress increased, while the mRNA level of DnaJB13 was downregulated at multiple time points during heat stress treatment. The mRNA levels of all four DnaJBs were upregulated by cold and UV stress. In addition, the expression levels of DnaJB6, DnaJshv and DnaJB13 were reduced under abamectin, imidacloprid, cypermethrin, bifenthrin, spirodiclofen, and methomyl stresses. The transcript level of DnaJB14 was decreased by imidacloprid, cypermethrin, spirodiclofen, and methomyl exposure but increased by abamectin and bifenthrin exposure. These results indicate that the demand of A. cerana cerana for these four DnaJBs differs under various stress conditions. To further explore the role of DnaJBs in the stress response, we successfully silenced DnaJshv and DnaJB14. The content of protein carbonyl was increased, while the content of VC, the enzymatic activities of CAT, GST, and SOD, the mRNA levels of many antioxidant-related genes, and the total antioxidant capacity were reduced after knockdown of DnaJshv and DnaJB14 in A. cerana cerana. These results indicate that silencing DnaJshv and DnaJB14 increases oxidative damage and decreases the antioxidant ability of A. cerana cerana. Taken together, our results demonstrate that DnaJB6, DnaJshv, DnaJB13, and DnaJB14 are differentially expressed under stress conditions and play crucial roles in response to various stressors, possibly through the antioxidant signalling pathway. These findings will be conducive to understanding the molecular basis of bee responses to environmental stresses and are beneficial for improving bee protection.

作为关键传粉者，蜜蜂常暴露于多重环境胁迫之中，并已演化出适应这些胁迫因子的关键机制。然而，其基因层面介导的分子基础仍有待揭示。本研究从中华蜜蜂（Apis cerana cerana）中鉴定得到4个热休克蛋白DnaJB亚家族基因：DnaJB6、DnaJshv、DnaJB13与DnaJB14，上述基因的蛋白序列中均含有J结构域。DnaJB6与DnaJshv的表达水平可被热胁迫不同程度地上调；DnaJB14的转录水平随热胁迫程度加深逐渐上调；而DnaJB13的mRNA水平在热胁迫处理后的多个时间点均呈下调趋势。冷胁迫与紫外（UV）胁迫均可上调4个DnaJBs的mRNA水平。此外，经阿维菌素、吡虫啉、氯氰菊酯、联苯菊酯、螺螨酯及灭多威胁迫处理后，DnaJB6、DnaJshv与DnaJB13的表达水平均有所降低。DnaJB14的转录水平在吡虫啉、氯氰菊酯、螺螨酯与灭多威处理后下降，但在阿维菌素与联苯菊酯处理后则呈上升趋势。上述结果表明，中华蜜蜂对这4个DnaJBs的需求在不同胁迫条件下存在差异。为进一步探究DnaJBs在胁迫响应中的作用，本研究成功对DnaJshv与DnaJB14进行了基因沉默。在中华蜜蜂体内沉默DnaJshv与DnaJB14后，其羰基化蛋白含量升高，而抗坏血酸（VC）含量、过氧化氢酶（CAT）、谷胱甘肽S-转移酶（GST）与超氧化物歧化酶（SOD）的酶活性、多个抗氧化相关基因的mRNA水平及总抗氧化能力均出现下降。上述结果表明，沉默DnaJshv与DnaJB14会加剧中华蜜蜂的氧化损伤并降低其抗氧化能力。综上，本研究结果证实，DnaJB6、DnaJshv、DnaJB13与DnaJB14在不同胁迫条件下呈差异表达，并在应对各类环境胁迫的过程中发挥关键作用，其机制可能与抗氧化信号通路相关。本研究结果有助于阐明蜜蜂响应环境胁迫的分子基础，同时可为蜜蜂保护工作提供理论支撑。