Table_1_RNA-Seq Analysis of Magnaporthe grisea Transcriptome Reveals the High Potential of ZnO Nanoparticles as a Nanofungicide.xlsx

Magnaporthe grisea is one of the most destructive pathogen that encounters a challenge to rice production around the worldwide. The unique properties of ZnO nanoparticles (NPs), have high attractiveness as nanofungicide. In the present study, the response of fungi to ZnO NPs was evaluated using RNA sequencing (RNA-seq). Two different aligners (STAR and Hisat2) were used for aligning the clean reads, and the DEseq2 package was used to identify the differentially expressed genes (DEGs). In total, 1,438 and 761 fungal genes were significantly up- and down-regulated in response to ZnO NPs, respectively. The DEGs were subjected to functional enrichment analysis to identify significantly enriched biological pathways. Functional enrichment analysis revealed that “cell membrane components,” “ion (calcium) transmembrane transporter activity,” “steroid biosynthesis pathway” and “catalytic activity” were the contributed terms to fungal response mechanisms. The genes involved in aflatoxin efflux pumps and ribosome maturation were among the genes showing significant up- and down-regulation after ZnO NPs application. To confirm the obtained RNA-seq results, the expression of six randomly selected genes were evaluated using q-RT-PCR. Overall, the RNA-seq results suggest that ZnO NPs primarily act on the fungal cell membrane, but accumulation of ROS inside the cell induces oxidative stress, the fungal catalytic system is disrupted, resulting into the inhibition of ROS scavenging and eventually, to the death of fungal cells. Our findings provide novel insights into the effect of ZnO NPs as a promising nanofungicide for effective control of rice blast disease.

稻瘟病菌（Magnaporthe grisea）是全球范围内对水稻生产造成严重威胁的毁灭性病原菌之一。氧化锌纳米颗粒（ZnO NPs）凭借其独特的理化特性，作为纳米杀菌剂展现出极高的应用潜力。本研究采用RNA测序（RNA-seq）技术，探究了稻瘟病菌对氧化锌纳米颗粒的响应机制：使用STAR与Hisat2两款序列比对工具对清洁读段进行比对，并通过DESeq2分析包筛选差异表达基因（DEGs）。经统计，共有1438个真菌基因在氧化锌纳米颗粒处理后显著上调表达，761个基因显著下调表达。研究人员对筛选得到的差异表达基因开展功能富集分析，以鉴定显著富集的生物学通路。功能富集结果显示，“细胞膜组分”、“离子（钙）跨膜转运活性”、“类固醇生物合成通路”与“催化活性”为参与稻瘟病菌响应氧化锌纳米颗粒的核心富集条目。参与黄曲霉毒素外排泵及核糖体成熟过程的基因，均在氧化锌纳米颗粒处理后呈现显著的表达差异。为验证RNA-seq的分析结果，本研究随机选取6个基因，采用实时定量反转录PCR（q-RT-PCR）对其表达量进行验证。综合来看，RNA-seq分析结果表明，氧化锌纳米颗粒主要作用于真菌细胞膜；细胞内活性氧（ROS）的积累会诱导氧化应激，破坏真菌的催化系统，进而抑制活性氧的清除过程，最终导致真菌细胞死亡。本研究结果为氧化锌纳米颗粒作为极具潜力的纳米杀菌剂，用于高效防治水稻稻瘟病提供了全新的理论支撑。