| Drought stress is one of the main factors affecting the normal growth and production of wheat and it is significantly important to study the mechanism responding to drought stress.In order to analyze the ABA-dependent and-independent signaling pathways in response to drought stress,The wheat cultivar “Aikang 58”,was selected as material to identify drought-responding genes using high-throughput sequencing technology,furthermore,the genes were analyzed with functional annotation and metabolic pathway enrichment analysis,and the main results are as follows:1.Ten genes that responded to drought stress were selected randomly and the transcriptional expression level of each gene was measured by q RT-PCR.It was found that there was a high correction with the results between qRT-PCR and RNA-seq,which indicated that the transcriptome sequencing data was reliable to be used for further analysis.2.Through high-throughput sequencing technology,the gene expression levels of wheat seedlings under six treatments were measured.The results showed that each group of samples obtained more than 68,687,164 raw reads,and the total data volume of all samples reached 180 G.By assembling and further screening genes responding to drought stress,4327 genes(log2 Fold >2)were found at 3h,of which 1338 genes belong to the ABA-dependent signaling pathway and2989 genes belong to the non-ABA-dependent signaling pathway;at 24 h A total of 5364 genes were discovered,of which 1943 genes belonged to ABA-dependent signaling pathways and 3421 genes belonged to non-ABA-dependent signaling pathways.KEGG metabolic pathway analysis found that the accumulation of more genes in the metabolic pathways are endoplasmic reticulum protein process,plant hormone signal transduction,plant pathogen interaction pathway,glutathione metabolism pathway.3.A total of 1326 up-regulated genes responding to drought stress were found in a rapid process.These genes were enriched in 90 metabolic pathways,among which plant pathogen interaction pathways had enriched 90 genes and they were mainly found in CaM/CML;56up-regulated genes were found in glutamate metabolic pathways and they distributed in both ABA-dependent and-independent signaling,and the up-regulated expression genes are mainly enriched in GST;in addition,a large number of plant hormones respond to drought stress at 3 h,such as auxin,abscisic acid,ethylene,jasmine Acid,salicylic acid and so on.4.A total of 5364 up-regulated genes responding to drought stress were found at 24 h,of which 1943 and 3421 up-regulated genes were present in ABA-dependent and –independent signaling pathways.KEGG analysis found that a total of 1702 genes were mapped to 101 metabolic pathways,of which 54 genes were found in the plant pathogen interaction pathway,and they were mainly enriched in Ca M/CML;70 up-regulated genes were enriched in the glutathione pathway and they were mainly enriched in GST.5.In this study,a total of 1889 genes were found to up-regulated expression continuously and significantly and they were mapped to 70 metabolic pathways.The protein process on the endoplasmic reticulum was enriched with a large number of genes,in which the genes were mainly involved in ERAD and ubiquitin ligase complexes.synthesis.In addition,GST,CaM/CMLs,and plant hormones(auxin,ethylene,ABA,JA,etc.)all played an important role in response to drought stress.6.Transcription factors play an important regulatory role in response to drought stress in plants.In this study,a total of 55 kinds of transcription factors were found,and they distributed in both ABA-dependent and-independent signaling pathways.WRKY transcription factor enriched the most number of genes,78 and 26 genes were found at 3 h and 24 h respectively.In addition,AP2-EREBP,MYB,C2H2,NAC,bHLH and other transcription factors were all play an important role in response to drought stress. |
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