The transcription factor bZIP60 links the Unfolded Protein Response (UPR) to the Heat Stress Response (HSR) in maize
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ABSTRACT: This was a comparative transcriptome analysis by using high throughput sequencing. To assess the effects of heat stress on maize we used a controlled environment facility called the Enviratron to simulate field conditions. For our experiments, maize plants were subjected to conditions simulating normal diurnal rhythms of light and temperature, with increasing maximal daily temperature (MDT). Maize plants were grown continuously under four different temperature regimes with simulated morning temperatures ramped up over 6 hr to the MDT of 31°C, 33°C, 35°C or 37°C and simulated evening/night time temperatures ramped down over 8 hr to 10°C below the MDT. We tracked the gene expression events of maize W22 seedlings grow under different temperatures (MDT of 31°C, 33°C, 35°C or 37°C) to evaluate how different MDTs affect the program of gene expression in maize. At the same time, we analyzed the effects of temperature on gene expression in bzip60-2 and W22 V4 plants (20 DAG) and V5 plants (27 DAG) in the Enviratron as the temperature reached its MDT to investigate whether and how bZIP60 confers heat stress tolerance in maize. RNA was extracted from small strips of leaf lamina excised from the first fully expanded leaf of V4 and V5 W22 plants (at 20 and 27 DAG, respectively). Plants were sampled in triplicates.
Project description:This was a comparative transcriptome analysis by using high throughput sequencing. To assess the effects of heat stress on maize alternative splicing we used a controlled environment facility called the Enviratron to simulate field conditions. For our experiments, maize plants were subjected to conditions simulating normal diurnal rhythms of light and temperature, with increasing maximal daily temperature (MDT). Maize plants were grown continuously under four different temperature regimes with simulated morning temperatures ramped up over 6 hr to the MDT of 31°C, 33°C, 35°C or 37°C and simulated evening/night time temperatures ramped down over 8 hr to 10°C below the MDT. We tracked the alternative splicing events of maize W22 seedlings grow under different temperatures (MDT of 31°C, 33°C, 35°C or 37°C) to evaluate how different MDTs affect the program of gene alternative splicing in maize. RNA was extracted from small strips of leaf lamina excised from the first fully expanded leaf of V4 and V5 W22 plants (at 20 and 27 DAG, respectively). Plants were sampled in triplicates.
Project description:We sequenced mRNA from from four segments along a developing 9 day old third maize leaf of WT W22 accession and ZmDCT2 KO (in W22 background - dct2-1::Ac) plants, to identify changes in gene expression in dct2-1::Ac as a response to the loss of ZmDCT2 function.
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:Maize is a staple food, feed, and industrial crop. Heat stress (HS) is one of the major stresses affecting maize production and is usually accompanied by other stresses, such as drought. Our previous study identified a heterotrimer complex, ZmNF-YA1-YB16-YC17, in maize. ZmNF-YA1 and ZmNF-YB16 were positive regulators of the drought stress response and were involved in maize root development. In this study, we investigated whether ZmNF-YA1 confers HS tolerance in maize. The ZmNF-YA1 mutant and overexpression lines were used to test the role of ZmNF-YA1 in maize thermotolerance. The ZmNF-YA1 mutant was more temperature-sensitive than the WT, while the ZmNF-YA1 overexpression lines showed a thermotolerant phenotype. Higher malondialdehyde content and reactive oxygen species accumulation were observed in the mutant, followed by WT and overexpression lines after HS treatment, while an opposite trend was observed for chlorophyll content. RNA-seq was used to analyze transcriptome changes in W22 and nf-ya1 plants in response to HS at great depths. Based on their expression profiles, the HS response-related differentially expressed genes in nf-ya1 and W22 were grouped into seven clusters via k-means clustering. Gene Ontology (GO) enrichment analysis of Clade 1, in which ZmNF-YA1 and HS induced, GO terms for protein refolding and protein stabilization, and terms for various stress responses were considerably enriched. Thus, the contribution of ZmNF-YA1 to protein stabilization, refolding, and regulation of ABA, ROS, and heat/temperature signaling may be the major reason why ZmNF-YA1 overexpression enhanced heat tolerance, and the mutant showed a heat-sensitive phenotype.
Project description:To determine the role of MDT-15 in mRNA expression changes at different temperatures, we sequenced mRNAs obtained from day 1 adult Caenorhabditis elegans. Bristol N2 (wild-type) and mdt-15(tm2182) mutant animals that cultured at 25oC or 15oC were used.
Project description:These samples are from RNA collected from specimens held at 4 temperatures (15.5, 20.0, 25.0 and 28.6degC). Specimens were ramped to one of those temperatures over 4 days following a common garden at 11degC for two weeks. RNA was extracted from thorax tissue and used for RNA-seq. Genes with statistically different expression levels among temperatures were identified.
Project description:Analysis of whole genome bisulfite data for 3 maize inbred lines (B73, PH207, and W22) with data aligned to the corresponding genome for determination of methylation level (CG, CHG, and CHH) across 100bp windows of the maize genome.
Project description:This was a pilot project carried out by Dr Wojciech Majeran to determine the maize pollen proteome harvested from field-grown W22 (T43) plants in plots on the Musgrave Research Farm (Cornell CALS) in Aurora (NY). To enhance proteome coverage, the pollen were separated into soluble and membrane bound protein fractions, and separated by SDS-PAGE followed by in-gel digestion and shot-gun proteomics using a nanoLC-Orbitrap system.