Transcriptome analysis of hrf1 transformed rice revealed crosstalk between disease resistance and drought tolerance mediated by Harpin
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ABSTRACT:
ORGANISM(S): Oryza sativa
PROVIDER: GSE36661 | GEO | 2013/12/29
SECONDARY ACCESSION(S): PRJNA153669
REPOSITORIES: GEO
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Project description:Harpin proteins from Gram-negative plant pathogenic bacteria can activate distinct signaling pathways and cause multiple effects in plants. In this document, we observed the transcriptome profile in transgenic NJH12 and wide type R109 using a customized 57 k rice cDNA microarray and found 225 differentially expressed transcripts related to the responses to biotic and abiotic stresses, including programmed cell death (PCD), defense responses associated with the recognition of pathogen-derived elicitors secondary metabolite biosynthesis, and defense signalling pathways mediated by abscisic acid (ABA), reactive oxygen species (ROS), nitric oxide (NO), polyamines (PAs) and Ca2+, suggesting that the disease resistance and drought tolerance mediated by Harpin protein in hrf1-expressing NJH12 rice has a crosstalk through several signaling pathways. The homozygous T3 NJH12 line is a transgenic rice line generated by transforming rice cultivar R109 with hrf1-containing pBMH9 using Agrobacterium-mediated transformation (Shao et al., 2008). Three T3 progeny of the transgenic NJH12 used in this study were grown in a sterile incubator at 28 ℃ in the greenhouse with a 16 h light/8 h dark cycle. Rice leaves were collected at the four-leaf stage and stored in liquid nitrogen prior to RNA extraction. Wild-type rice R109 was used as the negative control. The experiments for each transgenic line and wild type were repeated three times by three independently grown sets of plants for preparing RNA samples used for independent hybridization to the microarrays. Total RNA was extracted with TRIZOL reagent (Invitrogen, Gaithersburg, MD). The RNA quality was assessed by formaldehyde agarose gel electrophoresis and concentration was quantitated by spectrophotometric analysis. Total RNA was kept at −80 ℃ and sent to the Affymetrix core facility, where quality-control analysis was carried out before cDNA synthesis from the mRNA, labeling (through synthesis of cRNA with incorporation of biotinylated ribonucleotide analogs), and hybridization to the GeneChip Rice Genome Array (Affymetrix). This chip contains 57, 381 probe sets to query 51, 279 transcripts, which was designed based on the 59, 712 gene predictions of TIGR’s osa1 version 2.0 release. The chip was washed and then stained with streptavidin-phycoerythrin on an Affymetrix Fluidics Station 400, followed by scanning on a GeneChip Scanner 3000 (Affymetrix). The results were quantified and normalized using MicroArray Suite 5.0 (GCOS1.4) software (Affymetrix). The gene annotation and pathway identification were performed according to the method described by Shi et al. (2006) and Miao et al. (2010b).
2013-12-29 | E-GEOD-36661 | biostudies-arrayexpress
Project description:Crop breeding for board-spectrum disease resistance is the most profitable strategy to control diseases. However, knowledge on genes and mechanism of board-spectrum resistance is very limited. We fund a rice mutant ebr1 (enhanced blight and blast resistance 1) showed broad-spectrum resistance to both bacterial Xanthomonas oryzae pv. oryzae and fungal Magnaporthe oryzae associated with spontaneous programmed cell death (PCD), autoimmunity activation and stunting. EBR1 is located at the chromosome centromere and encodes a previously unknown RING-type protein with E3 ubiquitin ligase activity. Extensive yeast two-hybrid screening identified one EIP (EBR1-interacting) protein, EIP1, which belongs to the BAG (Bcl-2 associated athanogene) family that have been associated with stress responses and modulation of cell death.cross kingdoms. We showed that EIP1 is a specially target of EBR1 and is a key player activating PCD and immunity including the salicylic acid and jasmonate defense pathways in rice. Its function is suppressed EBR1-mediated degradation through ubiquitination. EIP1-RNAi could restore the PCD and disease resistance phenotypes of ebr1, while EIP1 overexpression caused strong PCD and enhanced resistance to rice pathogens likely ebr1. Together, our study reveals a unique E3 ligase-BAD protein module that orchestrates autoimmunity and trade-off between defense and growth in rice.
2016-11-17 | GSE81108 | GEO
Project description:PAMP (Pathogen-Associated Molecular Pattern) recognition plays an important role in innate immune responses both in plants and animals. Lipopolysaccharides (LPS) of gram-negative bacteria are a typical PAMP molecule and have been reported to induce defense-related responses such as suppression of hypersensitive responses, defense gene espression and systemic resistance in plant. However the detailed analysis of these cellular responses and the molecular machinery involved in the perception and transduction of LPS molecule largely remains to be studied. Furthermore, the biological activities of LPS on plants have so far been reported only for dicots and no information is available on the action of LPS on monocots. We report here that bacterial LPSs, both from plant pathogens and non-pathogens, could induce various defense responses in rice cells, including reactive oxygen generation and defense gene expression. Global analysis of gene expression induced by two PAMP elicitors, LPS and chitin oligosaccharide elicitor, showed a close correlation between the gene responses induced by these elicitors, indicating the convergence of signaling cascades downstream of corresponding receptors. Further, we show that the defense responses induced by LPS are associated with programmed cell death, a finding so far not reported for LPS action on plant cells. Keywords: elicitor, defense, LPS, rice cell
2006-09-28 | GSE5906 | GEO
Project description:Seedlings blight is one of the destructive diseases of rice, caused by the fungus Fusarium oxysporum, which impairs rice productions. Fluoro-substituted benzothiadiazole derivatives (FBT) and chitosan oligosaccharide (COS) are elicitors that can enhance plant resistance to pathogen infection. However, there is a lack of information regarding FBT and COS used as elicitors in rice seedlings blight. Therefore, the aim of this study was to evaluate the effect of FBT and COS treatments against rice seedling blight and elucidate the possible molecular mechanisms of the two elicitors for inducing resistance by proteomics. The results showed that FBT and COS significantly reduced the disease incidence and index, and improved the growth status of rice root caused by F. oxysporum. Biochemical analysis demonstrated that the two elicitors effectively enhanced activities of defense enzymes. Moreover, proteomic profiling analysis of rice root tissues disclosed more differentially expressed proteins in diterpenoid biosynthesis pathway induced by the two elicitors compared with other biological pathways, resulting in the accumulation of antimicrobial substance--momilactone. This study provided the basis of theory and application for FBT and COS used as rice elicitors against seedling blight on roots.
2019-08-12 | PXD014979 |
Project description:PAMP (Pathogen-Associated Molecular Pattern) recognition plays an important role in innate immune responses both in plants and animals. Lipopolysaccharides (LPS) of gram-negative bacteria are a typical PAMP molecule and have been reported to induce defense-related responses such as suppression of hypersensitive responses, defense gene espression and systemic resistance in plant. However the detailed analysis of these cellular responses and the molecular machinery involved in the perception and transduction of LPS molecule largely remains to be studied. Furthermore, the biological activities of LPS on plants have so far been reported only for dicots and no information is available on the action of LPS on monocots. We report here that bacterial LPSs, both from plant pathogens and non-pathogens, could induce various defense responses in rice cells, including reactive oxygen generation and defense gene expression. Global analysis of gene expression induced by two PAMP elicitors, LPS and chitin oligosaccharide elicitor, showed a close correlation between the gene responses induced by these elicitors, indicating the convergence of signaling cascades downstream of corresponding receptors. Further, we show that the defense responses induced by LPS are associated with programmed cell death, a finding so far not reported for LPS action on plant cells. Experiment Overall Design: 1. LPS treatment (WT), 2. LPS treatment (WT) color-swap, 3. N-acetylchitooctaose treatment (WT), 4. N-acetylchitooctaose treatment (WT) color-swap
2008-11-06 | E-GEOD-5906 | biostudies-arrayexpress
Project description:High-throughput sequencing of small RNAs from rice was used to identify distinct miRNAs that are responsive to elicitors from the fungal pathogen Magnaporthe oryzae. [Expression profiling by array] We used microarrays to determine the expression behaviour of target genes for elicitor-regulated miRNAs. [High throughput sequencing] High-throughput sequencing of rice small RNAs was performed in two different tissues, leaves and roots, and two different time point of elicitor treatment, 30' and 2h Amplicons were prepared by 5M-BM-4and 3M-BM-4adaptor ligation in which the 5'-adaptor contained a 'barcode' consisting of a 4-nucleotide identifier sequence for each sample. The libraries containing unique barcodes were combined and subjected to pyrosequencing (454 Life SciencesTM, Roche) [Expression profiling by array] Leaves from rice plants were harvested at two time points after the onset of treatment (30' and 2h) with elicitors of Magnaporthe oryzae 18.1 and used for RNA extraction and hybridization on Affymetrix microarrays. Mock inoculations were performed with sterile water for control experiments. Three biological replicates were analyzed. Each sample represented a pool of approximately 150 rice plants. [High throughput sequencing] 8 samples examined: leaves and roots, treated or not with elicitors at two different time points, 30' and 2h (2x2x2)
2014-01-15 | E-GEOD-30583 | biostudies-arrayexpress
Project description:Rice in tropical and sub-tropical areas is often subjected to cold stress at the seedling stage resulting in poor growth and yield loss. In general, japonica rice has stronger cold tolerance (CT) than indica rice. However, several favorite alleles for CT exist in indica rice and can be used to enhance CT under japonica background. Genome-wide gene expression profiling is the efficient way to decipher the molecular genetic mechanism of CT enhancement and provide valuable information for the CT improvement in rice molecular breeding. In this study, the transcriptome of a CT introgressed line (IL) K354 and its recurrent parent C418 under cold stress were comparatively analyzed to explore the possible CT enhancement mechanism of K354.Totally 3184 differentially expressed genes (DEGs) including 195 transcription factors were identified in both lines under cold stress, about half of them were commonly regulated, which were involved in the major cold responsive pathways including OsDREB1s regulon. The K354-specific cold-induced DEGs were mainly related to stimulus response, cellular cell wall organization, and microtubule-based movement process compared with commonly cold-induced DEGs by GO analysis. Moreover, 296 constitutive DEGs with significantly different transcription level between C418 and K354 were detected under both control and cold stress conditions. K354-specific cold-regulated and constitutive DEGs jointly account for the CT improvement of K354. Pathway analysis unraveled up-regulation of starch and sucrose metabolism in both genotypes and presumably weaker defense response to stress in K354 than C418 under cold stress. Candidate genes prediction based on previous putative CT genetic networks revealed genotype-dependent CT enhancement mechanism in CT IL K354 vs recurrent parent C418, including Sir2, OsFAD7, OsWAK112d, and PCD related genes, etc.We propose a hypothesis of the CT enhancement mechanism in rice based on the results in present study. Firstly, a number of cold-regulated genes are able to express constitutively at high level or absent under control condition for standing by the arrival of cold stress. Next, under cold stress slower perception of stress signal from the more cold-tolerant membrane can postpone the activation of defensive system which may have possible negative effects on rice growth. Then, PCD will be launched to sacrifice a few cells for maintaining the basal growth of most cells. Finally, the protective responses on multiple aspects of cold damage will be postponed because of delayed several cold-defensive pathways (i.e. OsDREB1C regulon). It might explain why the recovery capacity of K354 from cold stress to control condition is stronger than C418. The CT enhancement mechanism can be regarded as the possible way to improve CT of japonica rice using indica germplasm in rice breeding program. In this study, the specific gene expression patterns of two genotypes (C418 and K354) at 2h, 6h, 12h, 24h and 48h during cold stress treatment (4 C) and control were characterized by using the Affymetrix rice microarray platform.
2012-12-31 | E-GEOD-37940 | biostudies-arrayexpress
Project description:Arabidopsis thaliana cell suspension cultures (ACSC) were subjected to 30-min, mild chemical treatments with three different singlet oxygen elicitors at low-medium light conditions (150 µE m–2 s–1) with the aim of getting a better understanding of singlet oxygen-mediated defence responses in plants. The three elicitors Indigo Carmine (IC), Methylene Violet (MV) and Rose Bengal (RB) at a concentration of 0.5 µM were chosen because they exhibited different abilities to permeate the plasma membrane and to accumulate in the cell soma or organelles such as chloroplasts. In addition, ACSC were treated with 500 µM H2O2 for comparison. Confocal image analysis of Arabidopsis cells revealed that IC was not retained in cells, whereas MV and RB permeated the plasma membrane and accumulated in the chloroplast envelope and inside chloroplasts, respectively. As a consequence of their different cellular location, the physiological, transcriptional and photosynthetic responses of Arabidopsis cells to singlet oxygen production varied from each other and the activation of programmed cell death (PCD) was observed in ACSC treated with 0.5 µM RB, but not with the other elicitor nor with 500 µM H2O2. The role of chloroplasts in the activation of PCD was further investigated when this physiological response was analyzed in dark-grown cell cultures containing undifferentiated plastids. Interestingly, PCD was only activated in light-grown, but not in dark-grown, Arabidopsis cell cultures, suggesting that singlet oxygen-mediated defence responses were initiated inside chloroplasts. Genome-wide transcriptional profile analyses were performed as well and the results proved that there were only statistically significant changes in the transcript expression of light-grown ACSC treated with 0.5 µM RB and 500 µM H2O2, but not with IC nor with MV. Functional enrichment analyses revealed that GO/Biological process terms associated with defence responses were common in the treatments with 0.5 µM RB and 500 µM H2O2; however, resistance response to pathogen and PCD terms were only significantly over-represented in the RB treatment. Moreover, the analysis of the up-regulated transcripts in ACSC treated with 0.5 µM RB brought out that both specific markers for singlet oxygen from the conditional fluorescence (flu) mutant of Arabidopsis and transcripts with a key role in hormone-activated PCD (i.e. ethylene and jasmonic acid) were present, although there was no evidence for the up-regulation of EDS1 encoding the ENHANCED DISEASE SUSCEPTIBILITY PROTEIN 1. Finally, a co-regulation analysis proved that ACSC treated with 0.5 µM RB exhibited higher correlation with the flu family mutants than with other singlet oxygen producer mutants of Arabidopsis or wild-type plants of Arabidopsis subjected to high light treatments, where singlet oxygen was produced in photosystem II and an acclimatory response was activated instead of PCD. 18 Samples total. Six Samples are controls, 3 Samples each treated with Rose Bengal, Hydrogen Peroxide, Methyl Violet, and Indigo Carmine.
2014-03-13 | E-GEOD-43551 | biostudies-arrayexpress
Project description:Insect elicitors, in particular fatty acid amides like volicitin, have been known to induce defense-related gene expression. Here we investigated transcriptional changes in response to volicitin 60min after treatment locally and in distal parts of the treated leaf.
2013-09-19 | GSE50981 | GEO
Project description:Overexpression of programmed cell death 5 gene (PDCD5) in tumor cells enhances apoptosis triggered by growth factor or serum deprivation, and overexpression of its homolog, OsPDCD5, induces the early death of transgenic plants. In this work, a system of inducible OsPDCD5 expression using a heat shock promoter was developed to study PCD in rice at different developmental stages.The results showed that in three-leaf aged and older seedlings, OsPDCD5 could independently induce PCD. In altered plants, OsPDCD5 expression caused lesion mimic phenotype, abnormal leaf morphology, nuclear condensation, DNA fragmentation, and H2O2 production. But two-leaf aged and younger seedlings seedlings showed no visibly morphological phenotype after OsPDCD5 expression, suggested that young seedlings possessed some mechanism inhibiting OsPDCD5 induced PCD. The transcripts of 24-day-old transgenic Zhonghua 11 seedlings heat-shocked for 6 h (before the appearance of PCD phenotypes) were compared with untreated seedlings using microarray analysis (3-Leaf Group) to study the genes involved in OsPDCD5 induced PCD. The experiment was also performed using 18-day-old seedlings to discover why OsPDCD5 expression did not induce PCD in very young seedlings (2-Leaf Group). RNA samples were extracted using TRIzol (Invitrogen) as described by the manufacturer. Microarray analyses were carried out using a competitive hybridization method using the Agilent microarray system. All procedures were carried out according to the manufacturer's protocols. Briefly, total RNA from each sample was used to synthesize cRNA and was labeled with cyanine-5 (Cy5)- or cyanine-3 (Cy3)-labeled CTP. The labeled cRNAs (including a heat-shocked and a control sample) were competitively hybridized to the Agilent Rice Oligo Microarrays, and then washed. The hybridized slides were scanned using Agilent DNA Microarray Scanner and data points were extracted using Agilent Feature Extraction software. Two comparisons were made between biologically independent samples.
2011-06-30 | E-GEOD-18900 | biostudies-arrayexpress