ABSTRACT: Transcription profiling by array of seedlings from two maize inbred lines (Ye478 and Han21 )under drought stress, re-watering after drought, or normal watering conditions
Project description:To understand the transcriptome changes during drought tolerance in maize, the drought-tolerant line Han21 and drought-sensitive line Ye478, which show substantial differences in drought tolerance at the seedling stage, were selected for this study. Using the GeneChip Maize Genome Arrays, we applied genome-wide gene expression analysis to the two genotypes under gradual drought stress and re-watering. We identified 2172 common regulated transcripts in both lines under drought stress, with 1084 common up-regulated transcripts and 1088 common down-regulated transcripts. Among the 2172 transcripts, 58 potential protein kinases and 117 potential transcription factors were identified. The potential components of the ABA signaling pathway were identified from the common regulated transcripts. We also identified 940 differentially regulated transcripts between the two lines. Among the 940 transcripts, the differential expression levels of 29 transporters and 15 cell wall-related transcripts may contribute to the different tolerances of the two lines. Additionally, we found that the drought-responsive genes in the tolerant Han21 line recovered more quickly when the seedlings were re-watered, and 311 transcripts in the tolerant Han21 line were exclusively up-regulated at the re-watering stage compared to the control and stress conditions. Our study provides a global characterization of two maize inbred lines during drought stress and re-watering and will be valuable for further study of the molecular mechanisms of drought tolerance in maize.
Project description:To understand the transcriptome changes during drought tolerance in maize, the drought-tolerant line Han21 and drought-sensitive line Ye478, which show substantial differences in drought tolerance at the seedling stage, were selected for this study. Using the GeneChip Maize Genome Arrays, we applied genome-wide gene expression analysis to the two genotypes under gradual drought stress and re-watering. We identified 2172 common regulated transcripts in both lines under drought stress, with 1084 common up-regulated transcripts and 1088 common down-regulated transcripts. Among the 2172 transcripts, 58 potential protein kinases and 117 potential transcription factors were identified. The potential components of the ABA signaling pathway were identified from the common regulated transcripts. We also identified 940 differentially regulated transcripts between the two lines. Among the 940 transcripts, the differential expression levels of 29 transporters and 15 cell wall-related transcripts may contribute to the different tolerances of the two lines. Additionally, we found that the drought-responsive genes in the tolerant Han21 line recovered more quickly when the seedlings were re-watered, and 311 transcripts in the tolerant Han21 line were exclusively up-regulated at the re-watering stage compared to the control and stress conditions. Our study provides a global characterization of two maize inbred lines during drought stress and re-watering and will be valuable for further study of the molecular mechanisms of drought tolerance in maize. In two independent experiments, we generate maize gene expression profiles during drought stress and re-watering through comparing genome-wide expression patterns of drought stress treatment and re-watering treatment by using 17,555 Affymetrix maize whole genome array.
Project description:This was a comparative transcriptome analysis by using high throughput sequencing. To assess the effects of drought stress and NF-Y transcription factors ZmNF-YA1 and ZmNF-YB16 on maize, leaves from wild-type (W22), zmnf-ya1 (m67) mutant, wild-type (B104) and ZmNF-YB16 overexpression (OE) plants grow under well-watered and drought stress conditions were collected and RNAseq was performed. We tracked the gene expression events of inbred maize lines W22 or B104 seedlings in response to drought stress to evaluate how drought stress affects the gene expression program in maize. At the same time, we analyzed the effects of drought stress on gene expression in zmnf-ya1 and ZmNF-YB16 OE plants to investigate whether and how ZmNF-YA1 and ZmNF-YB16 confer drought stress tolerance in maize. Maize plants were grown under well-watered conditions until the V4 stage (zmnf-ya1 and W22) or V9 stage (ZmNF-YB16 OE and B104), and then half of them were exposed to drought stress treatment. Water loss in the soil and the electrolyte leakage from leaf cells were used to assess drought stress in plants. Leaves from 3-4 plants were pooled for each sample, and two replicates were used. RNA was extracted from small strips of leaf lamina excised from the first fully expanded leaf of the plants.
Project description:MicroRNAs (miRNAs) are a class of small, non-coding regulatory RNAs that regulate gene expression by guiding target mRNA cleavage or translational inhibition in plants and animals. At present there is relatively little information regarding the role of miRNAs in the response to drought stress in maize. In this study, two small RNA libraries were sequenced, and a total of 11,973,711 and 14,326,010 raw sequences were generated from growing leaves of drought-tolerant and drought-sensitive maize seedlings, respectively. Further analysis identified 192 mature miRNAs, which include 124 known maize (zma) miRNAs and 68 potential novel miRNA candidates. Additionally, 167 target genes (259 transcripts) of known and novel miRNAs were predicted to be differentially expressed between two maize inbred lines. Of these, three novel miRNAs were up-regulated and two were down-regulated under drought stress. The expression of these five miRNAs and nine target genes was confirmed using quantitative reverse transcription PCR. The expression of three of the miRNAs and their putative target genes exhibited an inverse correlation, and expression analysis suggested that all five may play important roles in maize leaves. Finally, GO annotations of the target genes indicated a potential role in photosynthesis, may therefore contribute to the drought stress response. This study describes the identification and characterization of novel miRNAs that are the differentially expressed in drought-tolerant and drought-sensitive inbred maize lines. This provides the foundation for further investigation into the mechanism of miRNA function in response to drought stress in maize.
Project description:To identify novel miRNA and NAT-siRNAs that are associated with abiotic stresses in maize, we generated small RNA sequences from maize seedlings that grew under control and under dought, salt, and cold stress treatments. Sequencing of small RNAs in maize under control, drought, salt, and cold stress conditions.
Project description:To understand the molecular mechanism of drought stress resistance mediated by OsABA8ox3 gene, we checked the genome-wide expression profile changes in the OsABA8ox3 RNAi and WT seedlings using the Affymetrix GeneChip under the normal condition and drought stress. A total of 1436 genes showed greater than 2-fold higher expression levels in both WT and RNAi-9 seedlings after drought stress, and most of them had higher up-regulated folds in RNAi-9 seedlings than that of WT. Gene expressions in the OsABA8ox3 RNAi and WT seedlings under the normal and drought stress conditions.
Project description:Drought is a major abiotic stress that threatens global food security. Circular RNAs (circRNAs) are endogenous RNAs. How these molecules influence plant stress responses remains elusive. Here, a large scale circRNA profiling identified 2174 and 1354 high-confidence circRNAs in maize and Arabidopsis, respectively, and most were differentially expressed in response to drought. A substantial number of drought-associated circRNA hosting genes were involved in conserved or species-specific pathways in drought responses. Comparative analysis revealed that circRNA biogenesis was more complex in maize than in Arabidopsis. In most cases, maize circRNAs were negatively correlated with sRNA accumulation. In 368 maize inbred lines, the circRNA-hosting genes were enriched for SNPs associated with circRNA expression and drought tolerance, implying either important roles of circRNAs in maize drought responses or their potential use as biomarkers for breeding drought-tolerant maize. Additionally, the expression levels of circRNAs derived from drought-responsible genes encoding calcium-dependent protein kinase and cytokinin oxidase/dehydrogenase were significantly associated with drought tolerance of maize seedlings. Specifically, Arabidopsis plants overexpressing circGORK (Guard cell outward-rectifying K+-channel) were hypersensitive to ABA, but insensitive to drought, suggesting a positive role of circGORK in drought tolerance. We report the transcriptomic profiling and transgenic studies of circRNAs in plant drought responses, and provide evidences highlighting the universal molecular mechanisms involved in plant drought tolerance.
Project description:Purpose: To study the effects of drought at the transcriptomic level on two different actively dividing maize tissue: the ovaries, and the leaf meristem Methods: The Illumina reads were mapped to the maize B73 reference genome using Tophat followed by transcriptome reconstruction using Cufflinks. The FPKM valuse were extracted from cufflinks output and an R package called Limma was used to identify differentially expressed genes under drought under both tissues Results and Conclusions: Different processes which were differentially expressed under drought in both tissues were identified and analyzed in detail. A working hypothesis was formulated to account for the observed susceptibility of the reproductive tissue when compared to the robust response of the vegetative tissue. This analysis also servers as a basis for future study on drought-induced embryo abortion. Maize (Zea mays) plants of inbred line B73 were grown in the green house under well watered and drought stress conditions until they reached the reproductive stage (at the onset of silk emergence). For the drought stress two to three days after irrigation was withheld, the plants were hand pollinated, and 24 hours after pollination measurements and samples were collected for transcriptome analysis. At the end of the drought period (1DAP) the basal leaf meristem of the three youngest leaves and the ovary tissues were sampled for Illumina deep sequencing. Samples were labeled as well watered control leaf meristem (MLC), well watered control ovaries/ "cob" (MCC), drought stressed leaf meristem (MLD) and drought stressed ovary tissue (MCD). There are 8 libraries in total including one biological replicate for each condition.
Project description:The mitogen-activated protein kinase (MAPK) cascade is an evolutionarily conserved signal transduction pathway that is involved in plant development and stress responses. As the first component of this phosphorelay cascade, mitogen-activated protein kinase kinase kinases (MAPKKKs) act as adaptors linking upstream signaling steps to the core MAPK cascade to promote the appropriate cellular responses; however, the functions of MAPKKKs in maize are unclear. Here, we identified 71 MAPKKK genes, of which 14 were novel, based on a computational analysis of the maize (Zea mays L.) genome. Using an RNA-seq analysis in the leaf, stem and root of maize under well-watered and drought-stress conditions, we identified 5,866 differentially expressed genes (DEGs), including 8 MAPKKK genes responsive to drought stress. Many of the DEGs were enriched in processes such as drought stress, abiotic stimulus, oxidation-reduction, and metabolic processes. The other way round, DEGs involved in processes such as oxidation, photosynthesis, and starch, proline, ethylene, and salicylic acid metabolism were clearly co-expressed with the MAPKKK genes. Furthermore, a quantitative real-time PCR (qRT-PCR) analysis was performed to assess the relative expression levels of MAPKKKs. Correlation analysis revealed that there was a significant correlation between expression levels of two MAPKKKs and relative biomass responsive to drought in 8 inbred lines. Our results indicate that MAPKKKs may have important regulatory functions in drought tolerance in maize.
Project description:To understand the molecular mechanism of drought stress resistance mediated by OsABA8ox3 gene, we checked the genome-wide expression profile changes in the OsABA8ox3 RNAi and WT seedlings using the Affymetrix GeneChip under the normal condition and drought stress. A total of 1436 genes showed greater than 2-fold higher expression levels in both WT and RNAi-9 seedlings after drought stress, and most of them had higher up-regulated folds in RNAi-9 seedlings than that of WT.