Data_Sheet_1_Transcription Analysis of the Stress and Immune Response Genes to Temperature Stress in Ostrinia furnacalis.xlsx

Ostrinia furnacalis is one of the most important pests on maize. O. furnacalis larvae are frequently exposed to the temperature challenges such as high temperature in summer and cold temperature in winter in the natural environment. High and low temperature stress, like any abiotic stress, impairs the physiology and development of insects. Up to now, there is limited information about gene regulation and signaling pathways related to the high and cold stress response in O. furnacalis. High-throughput sequencing of transcriptome provides a new approach for detecting stress and immune response genes under high and low temperature stresses in O. furnacalis. In the present study, O. furnacalis larvae were treated with the temperature at 8 and 40°C, and the responses of O. furnacalis larvae to the temperature stress were investigated through RNA-sequencing and further confirmation. The results showed that immune responses were up-regulated in larvae by the cold stress at 8°C while some stress response genes, such as HSP family, GST-2, Bax inhibitor and P450, were significantly increased at 40°C. Furthermore, quantitative real time polymerase chain reaction were performed to quantify the expression levels of immune related genes, such as PGRP-LB, antimicrobial peptides, lysozyme, serine protease and stress response genes such as small HSPs and HSP90, and the expression levels of these genes were similar to the RNA-seq results. In addition, the iron storage protein Ferritin was found to be involved in the response to temperature stress, and the changes of total iron concentration in the hemolymph were, in general, consistent with the expression levels of Ferritin. Taken together, our results suggested that the stress response genes were involved in the defense against the heat stress at 40°C, and the immune responses triggered by cold stress might provide protection for larvae from cold stress at 8°C. More interestingly, our results showed that during the responses to temperature stress, the total iron concentration in hemolymph regulated by Ferritin increased, which might help O. furnacalis in surviving the low and high temperature stress.

Ostrinia furnacalis 作为玉米作物上最为重要的害虫之一，其幼虫在自然环境中常遭受温度挑战，如夏季的高温和冬季的低温。此类高温与低温的胁迫，如同其他非生物胁迫，会损害昆虫的生理机能和发育进程。截至目前，关于 O. furnacalis 对高温和低温胁迫的基因调控与信号通路的信息仍十分有限。转录组的高通量测序为检测 O. furnacalis 在高温和低温胁迫下的应激和免疫反应基因提供了一种新的途径。在本研究中，通过对 O. furnacalis 幼虫进行 8°C 和 40°C 温度处理，并利用 RNA 测序技术对 O. furnacalis 幼虫对温度胁迫的响应进行深入研究，并进一步验证。研究结果表明，在 8°C 的低温胁迫下，幼虫的免疫反应被上调，而如 HSP 家族、GST-2、Bax 抑制剂和 P450 等应激反应基因在 40°C 时显著增加。此外，通过定量实时聚合酶链式反应（qRT-PCR）对与免疫相关的基因，如 PGRP-LB、抗菌肽、溶菌酶、丝氨酸蛋白酶以及应激反应基因如小 HSPs 和 HSP90 的表达水平进行量化，结果显示这些基因的表达水平与 RNA-seq 结果相似。此外，发现铁储存蛋白 Ferritin 参与了对温度胁迫的响应，血液淋巴中的总铁浓度变化总体上与 Ferritin 的表达水平相一致。综合以上结果，我们的研究提出，应激反应基因参与了抵御 40°C 的热胁迫，而由低温胁迫引发的免疫反应可能为幼虫在 8°C 的低温胁迫下提供保护。更有趣的是，我们的研究结果还显示，在应对温度胁迫的过程中，由 Ferritin 调节的血液淋巴中的总铁浓度增加，这有助于 O. furnacalis 应对低温和高温的胁迫。