RNA Sequencing Reveals that Endoplasmic Reticulum Stress and Disruption of Membrane Integrity Underlie Dimethyl Trisulfide Toxicity against Fusarium oxysporum f. sp. cubense Tropical Race 4

Abstract

Fusarium wilt of banana, a destructive disease that affects banana production, is caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4). In a previous study, we confirmed the strong inhibitory effects of Chinese leek (Allium tuberosum) on the incidence of this disease. Sulfur compounds are the primary antifungal constituents of Chinese leek. Among these, dimethyl trisulfide (DT) was the most abundant and exhibited the strongest inhibition of Foc TR4 growth and development. In the present study, the global gene expression profiles of Foc TR4 isolates treated with DT at 4,000-folds dilution (concentration of 1/4,000, v/v) for 1.5, 6, and 12 h were investigated by using RNA sequencing. The expression patterns of 15 DEGs were validated based on quantitative real-time PCR (qRT-PCR) assay. Untreated sample presented 2,556, 1,691, and 1,150 differentially expressed genes (DEGs) at 1.5, 6, and 12 h after the onset of the experiment, respectively, whereas DT-treated isolates presented 2,823, 3,546, and 6,197 DEGs. Based on Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, DEGs involved in endoplasmic reticulum (ER), glycosylation, and steroid biosynthesis were significantly inhibited by DT exposure. The similar expressional patterns of 15 DEGs between RNA-seq and qRT-PCR assays indicated the reliability of the RNA-seq data. In conclusion, ER stress related to glycosylation inhibition and damage to cell membrane integrity might contribute to the toxicity of DT against Foc TR4. As the results presented here evidenced changes in gene expression associated with DT exposure, which might be used to develop new approaches for controlling FWB.

The Impairment of Methyl Metabolism From luxS Mutation of Streptococcus mutans

“…The S. mutans strains were cultured overnight. The isolation, purification, and reverse transcription of total bacterial RNA into cDNA was performed as described previously (Zuo et al, 2017 ). qRT-PCR was performed using the Bio-Rad CFX96 system (Bio-Rad, Marnes-la-Coquette, France).…”

Antifungal effects of dimethyl trisulfide against Colletotrichum gloeosporioides infection on mango

“…Previous studies showed that DMTS together with the volatiles from Chinese leek could inhibit conidial germination of Fusarium oxysporum f.sp. cubense tropical race 4 ( Foc TR4), and DMTS displayed the strongest inhibition among volatiles (Zhang et al, ; Zuo et al, , ). Moreover, DMTS together with the volatiles emitted from Streptomyces globisporus JK‐1 exhibited strong inhibitory effects on conidial germination of P. italicum and Botrytis cinerea , and DMTS acted as the main active compound (Li et al, , ).…”

“…DMTS has shown strong inhibitory effects on the growth of Penicillium italicum and Fusarium oxysporum f.sp. cubense tropical race 4 (Li et al, ; Zuo, Zhang, Chen, Chen, & Huang, ). However, the toxicity of DMTS to C. gloeosporioides has not been previously demonstrated.…”

RNA Sequencing Reveals That Both Abiotic and Biotic Stress-Responsive Genes are Induced during Expression of Steroidal Glycoalkaloid in Potato Tuber Subjected to Light Exposure

“…Poly(A) mRNA was purified by using oligo(dT) magnetic beads and then digested into short fragments (approximately 200 bp). The RNA-Seq library preparation method were performed according to our previous method that was described by Zuo et al (2017) [26] and it included three replicates for each treatment. The amplified fragments were sequenced while using an Illumina HiSeq TM 2000.…”

“…PCR conditions were 95 °C for 15 min., followed by 40 cycles of 95 °C for 10 s, and annealing/extension at 60 °C for 30 s. The melting curve was determined for each sample. Relative gene expression was calculated while using the cycle threshold (Ct)2 −ΔΔCt method, as described by Zuo et al (2017) and Livak and Schmittgen (2001) [26,33]. Data from qRT-PCR analysis were expressed as means ±SD of three independent replicates.…”