Next Generation Sequencing Facilitates Quantitative Analysis of Physcion and Chrysophanol on cucumber powdery mildew Transcriptomes
Ontology highlight
ABSTRACT:
ORGANISM(S): Cucumis sativus
PROVIDER: GSE72034 | GEO | 2015/08/14
SECONDARY ACCESSION(S): PRJNA292785
REPOSITORIES: GEO
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Project description:In a previous study, we found that H2S alleviates salinity stress in cucumber by maintaining the Na+/K+ balance and by regulating H2S metabolism and the oxidative stress response. However, little is known about the molecular mechanisms behind H2S-regulated salt-stress tolerance in cucumber. Here, an integrated transcriptomic and proteomic analysis based on RNA-seq and 2-DE was used to investigate the global mechanism underlying H2S-regulated salt-stress tolerance. In total, 11 761 differentially expressed genes (DEGs) and 61 differentially expressed proteins (DEPs) were identified. Analysis of the pathways associated with the DEGs showed that salt stress enriched expression of genes in primary and energy metabolism, such as photosynthesis, carbon metabolism and biosynthesis of amino acids. Application of H2S significantly decreased these DEGs but enriched DEGs related to plant-pathogen interaction, sulfur-containing metabolism, cell defense and signal transduction pathways. Notably, changes related to sulfur-containing metabolism and cell defense were also observed through proteome analysis, such as Cysteine synthase 1, Glutathione S-transferase U25-like, Protein disulfide-isomerase and Peroxidase 2. We present the first global analysis of the mechanism underlying H2S regulation of salt-stress tolerance in cucumber through tracking changes in the expression of specific proteins and genes.
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Project description:Disease resistance is associated with a plant defense response that involves an integrated set of signal transduction pathways. Gene Expression studies were performed using Arabidopsis wild type seedlings treated with and without salicylic acid (SA). In this study we examined pathogenesis related genes are upregulated. We used Affymetrix GeneChip to study the genome wide gene expression with and without induction of defense by treatment with SA Experiment Overall Design: Arabidopsis seedlings Col-0 were grown for 9 days in controlled environment rooms at 22°C temperature and a photoperiod of 16 h light 100 µm m2 s_1 cool-white fluorescent illumination. After 9 days seedlings were transferred into water as a mock treatment and with the defense related signaling molecules salicylic acid (SA) for one day. The concentration of SA (Sigma, St. Louis) was 2mM and pH 6.5. RNA extracted from SA and mock treated seedling and hybridize with Affymetrix GeneChip to examine the change in plant defence gene expression due to induction of SA.
2009-02-28 | E-GEOD-14961 | biostudies-arrayexpress
Project description:Corynespora leaf spot (CSL), caused by Corynespora cassiicola, has become one of the most important foliar diseases of cultivated cucumber. However, the defense mechanisms of cucumber plants in response to C. cassiicola are still poorly understood. Here, proteins from resistant plants were analyzed using isobaric tags for relative and absolute quantification (iTRAQ). A total of 286 differentially expressed proteins were identified (P<0.05, ratio>1.2 or <0.83) at 6 and 24 h after pathogen inoculation in the resistant cucumber cultivar Jinyou 38. Some of the early responses to C. cassiicola infection were revealed, and four vital clues regarding the resistance of Cucumis sativus to cucumber CLS were discovered. First, the proteomic approach revealed the modulation of signaling pathways in resistant cucumber plants in response to C. cassiicola infection. Second, the plant immune system recognizes the pathogen and initiates the expression of basal immune response proteins, including those related to defense and stress responses, signal transduction, cell metabolism and redox regulation. Third, the common stress pathways were activated by C. cassiicola; in particular, mildew resistance locus O (MLO) proteins played a crucial role in the prevention of CLS. Fourth, the rapid activation of the carbohydrate and secondary metabolic pathways, the modification and reinforcement of cell walls, and the adjustment of the apoplectic environment to high-stress conditions were crucial in cucumber resistance to CLS disease. Overall, our data increase the knowledge of incompatible interactions between plants and pathogens and provide new insight into the contribution of molecular processes in cucumber to disease resistance.
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Project description:Cucumber (Cucumis sativus L.) is an economically important vegetable cultivated all over the world. Grafting can produce bloomless or sparse-bloom cucumber, which is welcomed by increasing consumers. Bloom granule is tine glandular hair, which is hard and rare studied on its formation and related genes. Mutifunctional RNA-seq is a recently developed analytical approach for transcriptome profiling via high-throughput sequencing and has been recently applied to a wide variety of organisms, which provide us reliable technical means detect bloom formation and related genes. In this study, we chose a cucumber inbred line (Shannong No.5) and two pumpkin rootstock lines as materials, and constructed four tested cucumbers, grew plants in “Yamazaki cucumber nutrient solution formula” prepared by deionized water, treated plants with or without 1.7mM potassium silicate 2 hours before collecting pericarp. Each treatment were duplicated twice.16 cDNA libraries were constructed from pericarp of a cucumber inbred line (own-rooted cucumber), C/C (self-grafted cucumber), M/C (More bloom, cucumber grafted onto “3225” rootstock) and L/C(Less bloom, cucumber grafted onto “3212” rootstock). We obtained 17,215,769~17,529,047 high quality reads, and 18,804~19,358 genes from each sample. All reads can be mapped to the cucumber genome (Version 2). By RPKM comparing, we got 38 comparing combinations with differentially expressed genes (DEGs), obtained 38 significantly expressed combinations by FDR≤0.001 and the absolute value of log2Ratio≥1 as the thresholds. These results suggest that there are many differences and genes expression mode among effects of grafting or added silicon. This study addresses a preliminary analysis and offers a foundation for future genomic research in the bloom formation of cucumber.
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Project description:Trichomes are the hair-like structures that are widely present on the surface of aerial organs and function in plant defense against biotic and abiotic stresses. Previous studies focus on the single cell trichomes in Arabidopsis and cotton, or multicellular glandular trichomes in tomato, but the developmental process and molecular mechanisms controlling multicellular non-glandular trichome development are largely neglected. Here, we extensively characterized the fruit trichome (spine) development in wild type cucumber and in a tiny branched hair (tbh) mutant that contains a spontaneous mutation and has hairless foliage and smooth fruit surface. Our data indicated that cucumber trichome was multicellular and non-glandular, with no branches or endoreduplication. Further, the major feature of cucumber trichome development was spine base expansion. Transcriptome profiling through Digital Gene Expression indicated that meristem-related genes and transcription factors were implicated in the fruit spine development, and polarity regulators were upregulated during spine base expansion. qRT-PCR verified the reliability of our RNA-SEQ data, and in situ hybridization confirmed the enriched expression of meristem regulators CUP-SHAPED COTYLEDON3 (CUC3) and STM (SHOOT MERISTEMLESS) , as well as the abaxial identity gene KANADI (KAN) in cucumber fruit spine. Together, our results suggest a distinct regulatory pathway involving meristem genes and polarity regulators in multicellular trichome development in cucumber. Using Digital Gene Expression technology to compare the genome-wide gene expression profiles in the fruit spines of wild type cucumber and the tbh mutant, as well as the fruit spines on fruits of 0.5cm and 1.6cm long, repectively. Two biological repelicates were generated for each tissue.
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Project description:Salinity seriously reduces the yield and quality of crops. Silicon (Si) has been widely reported to have beneficial effects on plant growth and development under salt stress. However, the mechanism is still poorly understood. In an attempt to identify genes or gene networks that may be orchestrated to improve salt tolerance of cucumber plants, we profiled the RNA-seq transcriptomes of both control and salt-stressed cucumber leaves in the presence or absence of added Si. The comparative transcriptome analysis revealed that Si plays an important role in shaping the transcriptome of cucumber: the expression levels of >1,000 genes (differtially expressed genes, DEGs) were changed in response to Si treatment as compared with the control, and these genes were mainly involved in ion transport, hormone and signal transduction, biosynthesis and metabolic processes, stress and defense responses, and antioxidant activity. Under salt stress, many genes functionally associated with metabolic processes and responses to environmental stimuli were strongly up- or down-regulated in their expressions. Si treatment showed a tendency that induced the transcriptomic profile of salt-stressed cucumber back to that of the control with large majority of Na down-regulated DEGs and about half of Na up-regulated DEGs being adjusted back to CT levels. This study provides a novel insight into the mechanism for Si-mediated alleviation of salt stress in plants at the transcriptome level, and it suggests that Si may act as an elicitor to precondition cucumber plants and induce salt tolerance.
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Project description:Trichomes are the hair-like structures that are widely present on the surface of aerial organs and function in plant defense against biotic and abiotic stresses. Previous studies focus on the single cell trichomes in Arabidopsis and cotton, or multicellular glandular trichomes in tomato, but the developmental process and molecular mechanisms controlling multicellular non-glandular trichome development are largely neglected. Here, we extensively characterized the fruit trichome (spine) development in wild type cucumber and in a tiny branched hair (tbh) mutant that contains a spontaneous mutation and has hairless foliage and smooth fruit surface. Our data indicated that cucumber trichome was multicellular and non-glandular, with no branches or endoreduplication. Further, the major feature of cucumber trichome development was spine base expansion. Transcriptome profiling through Digital Gene Expression indicated that meristem-related genes and transcription factors were implicated in the fruit spine development, and polarity regulators were upregulated during spine base expansion. qRT-PCR verified the reliability of our RNA-SEQ data, and in situ hybridization confirmed the enriched expression of meristem regulators CUP-SHAPED COTYLEDON3 (CUC3) and STM (SHOOT MERISTEMLESS) , as well as the abaxial identity gene KANADI (KAN) in cucumber fruit spine. Together, our results suggest a distinct regulatory pathway involving meristem genes and polarity regulators in multicellular trichome development in cucumber.
2014-07-01 | GSE49607 | GEO
Project description:Purpose: The goals of this study are using RNA-seq to obtain cucumber and Botrytis cinerea transcriptome changes during infection Methods: mRNA profiles of anti-infection samples and interaction sample were generate by deep sequencing,using Illumina Hiseq 2500. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: BurrowsâWheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRTâPCR validation was performed using SYBR Green assays Results: Using an optimized data analysis workflow,In total, 248,908,688 raw reads were generated; after removing low-quality reads and those containing adapter and poly-N, 238,341,648 clean reads remained to map the reference genome. There were 3,512 cucumber (differential expression genes) DEGs and 1,735 B. cinerea DEGs. GO enrichment and KEGG enrichment analysis were performed on these DEGs to study the interaction between cucumber and B. cinerea. To verify the reliability and accuracy of our transcriptome data, 5 cucumber DEGs and 5 B. cinerea DEGs were chosen for RT-PCR verification. Conclusions:To the best of our knowledge, this is the first analysis of large-scale transcriptome changes of cucumber during the infection of Botrytis cinerea. These results will increase our understanding of the molecular mechanisms of the cucumber defense Botrytis cinerea and may be used to protect plants against disasters caused by necrotrophic fungal pathogens. mRNA profiles of infection and anti-infection cucumber were generated by deep sequencing, using Illumina Hiseq 2500 .
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