Rna clean up kit 5

For RNA sequencing, mycelium samples were collected from the V8 medium after 4 days of culturation. Total RNA was isolated from the samples using Trizol reagent (Invitrogen) and an RNA Clean-Up Kit-5 (Zymo Research, R1016), following the manufacturer's instructions. The extracted mRNA was used to construct the cDNA library, and the library construction was sequenced on an Illumina Hiseq 4000 Platform (Lc-Bio Technologies, Hangzhou, China). The preprocessed RNA-Seq reads were mapped to the reference genome of P. capsici strain LT1534 (https://mycocosm.jgi.doe.gov/Phyca11/Phyca11.home.html) [59 (link)] using the HISAT package [60 (link)], and the mapped reads of each sample were assembled using the StringTie method [61 (link)]. The DEGs were identified from RNA-Seq data with the cut-off of the corrected p-value < 0.05, using log2foldchange greater than or equal to 1 as a threshold. Analyses of the biological information of DEGs were performed using an online database (http://geneontology.org/); analysis of the KEGG (Kyoto Encyclopedia of Genes and Genomes) was performed using an online database (www.genome.jp/kegg) as a reference.

Because Zhengdan 958 is a hybrid cross, we chose the cv. B73, which had its genome sequenced, as the material for transcriptomic analysis. Due to the maize cv. B73 dying when treated at 47°C, the high stress temperature was changed to 42°C and the treatment time was increased to 1 day. For transcriptomic analysis, samples of roots were collected from the maize seedlings (cv.B73) previously soaked in guvermectin or water under the described high temperature conditions, as well as the samples from roots that had room temperature treatments. Total RNA was isolated from the samples using Trizol reagent (Invitrogen) and RNA Clean-Up Kit-5 (Zymo Research, R1016), following the manufacturers’ instructions. The extracted mRNA was used to construct the cDNA library, and the library construction was sequenced on an Illumina Hiseq 4000 Platform (Lc-Bio Technologies, Hangzhou, China). The preprocessed transcriptomic reads were mapped to the reference genome of maize cv.B73 using the HISAT package, and the mapped reads of each sample were assembled using the StringTie method. The differentially expressed genes (DEGs) were identified from RNA-Seq data with a cut-off for the corrected p-value < 0.005, using log2foldchange greater than or equal to 1 as a threshold. Analyses of the biological information of DEGs were performed using an online database (http://geneontology.org/).