Project description:The purpose of this experiment was to study the effects of the bacterial enzyme ACC deaminase on the transcriptional changes within canola seedlings. Seedlings from seeds treated with the plant growth-promoting bacteria Pseudomonas putida UW4 which expresses a high level of ACC deaminase and its ACC deaminase-minus mutant were compared to untreated seedlings along with a transgenic line of canola expressing the ACC deaminase enzyme in the roots. ACC deaminase breaks down 1-aminocyclopropane-1-carboxylic acid, the biosynthetic precursor to the plant hormone ethylene, lowering ethylene levels and improving plant fitness. Plants treated with the ACC deaminase-containing bacteria and transgenic plants expressing ACC deaminase are more tolerant to a variet of stresses and this expression study helps to illuminate the pathways responsible for the growth promotion provided by the beneficial bacteria and the role of the enzyme itself.
Project description:The purpose of this experiment was to study the effects of a bacterial ACC deaminase transgene in the roots and its impact on nickel tolerance of canola. ACC deaminase breaks down 1-aminocyclopropane-1-carboxylic acid, the biosynthetic precursor to the plant hormone ethylene, lowering ethylene levels and improving plant tolerance to stress. Ethylene evolved during plant stress is thought to causes senescence and cell death and worsen stress symptoms. Transgenic plants expressing ACC deaminase from the plant growth-promoting bacteria Pseudomonas putida UW4 are more tolerant to heavy metals in the soil and this expression study helps to illuminate the pathways responsible for this tolerance.
Project description:Canola plants inoculated with plant growth-promoting bacteria either expressing ACC deaminase or not to determine the effect on plant gene expression using an Arabidopsis microarray.
Project description:Canola plants inoculated with plant growth-promoting bacteria either expressing ACC deaminase or not to determine the effect on plant gene expression using an Arabidopsis microarray. 3 replicates for each ACD+ and ACD- bacteria, each compared with untreated control.
Project description:We profiled the gene regulatory landscape of Brassica napus reproductive development using RNA sequencing. Comparative analysis of this nascent allotetraploid across the plant lifecycle revealed the contribution of each subgenome to plant reproduction. Global mRNA profiling across reproductive development revealed lower accumulation of C subgenome transcripts relative to the A subgenome. Subgenome-specific transcriptional networks identified distinct transcription factor families enriched in each of the A and C subgenome in early seed development. Analysis of a tissue specific transcriptome of early seed development revealed transcription factors predicted to be regulators encoded by the A subgenome are expressed primarily in the seed coat whereas regulators encoded by the C subgenome were expressed primarily in the embryo. Whole genome transcription factor networks identified BZIP11 as an essential regulator of early B. napus seed development. Knockdown of BZIP11 using RNA interference resulted in knockdown of predicted target genes, and a reproductive-lethal phenotype. Our data indicate that subgenome bias are characteristic features of the B. napus seed throughout its development, and that such bias might not be universal across the embryo, endosperm, and seed coat of the developing seed. We also find that examining transcriptional networks spanning both the A and C genomes of the whole B. napus seed can identify valuable targets for seed development research. We suggest that-omics level approaches to studying gene regulation in B. napus can benefit from both broad and high-resolution analyses.
Project description:Background: The biological control agent Pseudomonas chlororaphis PA23 is effective at protecting Brassica napus (canola) from the necrotrophic fungus Sclerotinia sclerotiorum via direct antagonism. Despite the growing importance of biocontrol bacteria in plant protection from fungal pathogens, little is known about how the host plant responds to bacterial priming on the leaf surface or about changes in gene activity genome-wide in the presence and absence of S. sclerotiorum. Results: PA23 priming of mature canola plants reduced the number of lesion forming petals by 90%. Global RNA sequencing of the host pathogen interface showed a reduction in the number of genes uniquely upregulated in response to S. sclerotiorum by 16-fold when pretreated with PA23. Upstream defense-related gene patterns suggest MAMP-triggered immunity via surface receptors detecting PA23 flagellin and peptidoglycans. Although systemic acquired resistance was induced in all treatment groups, a response centered around a glycerol-3-phosphate (G3P)-mediated pathway was exclusively observed in plants treated with PA23 alone. Activation of these defense mechanisms by PA23 involved mild reactive oxygen species production as well as pronounced thylakoid membrane structures and plastoglobule formation in leaf chloroplasts. Conclusion: Further to the direct antibiosis that it exhibits towards the pathogen S. sclerotiorum, PA23 primes defense responses in the plant through the induction of unique local and systemic defense regulatory networks. This study has shed light on the potential effects of biocontrol agents applied to the plant phyllosphere. Understanding these interactions will aid in the development of biocontrol systems as a viable alternative to chemical pesticides in the protection of important crop systems.
Project description:Clubroot disease, caused by Plasmodiophora brassicae Woronin results in severe yield losses in Brassica crops, including canola. Silicon (Si) mitigates several stresses and enhances plant resistance to phytopathogens. We investigated the effects of Si on clubroot disease symptoms in canola at two concentrations of Si (Si1.0 and Si0.5). In addition, the effects of Si on P. brassicae-induced gene expression, endogenous levels of phytohormones and metabolites were also studied. Si application reduced clubroot symptoms and improved plant growth under greenhouse conditions. Pathogen-induced transcript-level changes were affected by Si treatment to P. brassicae with genes related to antioxidant activity, phytohormone biosynthesis and signalling, nitrogen metabolism and secondary metabolism exhibiting differential expression. Endogenous levels of several phytohormones (e.g., auxin, cytokinin, salicylic acid and abscisic acid), amino acids and secondary metabolites (e.g., glucosinolates) were affected by Si. This is the first report that Si ameliorates clubroot symptoms and its possible mode of action.
Project description:Phytohormones control many vital biological processes within plants, including growth and development, senescence, seed setting, fruit ripening and innate immunity. These biological processes are controlled by specific combinations of multiple phytohormones. The three main phytohormones involved in plant innate immunity are salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). SA is produced in response to biotrophic and hemibiotrophic pathogens. Whereas, JA and ET are produced in response to necrotrophic pathogens. Pseudomonas aeruginosa is found ubiquitously within the environment, and has been shown to participate in both beneficial and pathogenic interactions with host plants depending on bacterial strain and environment. In this study, we found that Brassica napus (canola) seedlings infected with P. aeruginosa strain PA14 displayed symptoms of disease and had significant weight loss in both root and shoot. Our transcriptomic data revealed that many molecular processes involved in plant innate immunity were upregulated, whereas photosynthesis was downregulated.
Project description:Drought is one of the most severe stresses leading to retardation of plant growth and development and loss of crop yield. Here we examined the proteome changes of an important oil seed crop canola Brassica napus under drought stress over a 14 day period. Using iTRAQ LC-MS/MS, we identified 2,244 proteins expressed during drought stress. Among them, 412 proteins showed significant changes in abundance under stress, and 67, 243, 287, and 79 proteins were differentially expressed in 3rd, 7th, 10th, and 14th day of drought stress, respectively. Functional analysis of the 412 proteins indicated that the number of proteins associated with “Metabolism”, “Protein synthesis”, and “Signaling” decreased, while those related to “Photosynthesis” and “Stress and defense” increased in response to drought stress. In particular, the proteome profiles at the 7th and 10th day were similar to each other, although there were much more post-translational modifications (PTM) at the 10th day of drought. Interestingly, 286 of 2,244 proteins exhibited PTMs in response to drought stress, 82 of which were differentially changed in drought-stressed plants, and 60 were observed at the 10th day. Furthermore, comparison of protein expression changes with those of gene transcription showed that there was positive correlation in B. napus, although there were different patterns between transcripts and proteins at each time point. As drought stress prolongs, most of the protein abundance changes may be attributed to gene transcription, and PTMs clearly contribute to the protein diversity and functions.
Project description:Sclerotinia sclerotiorum, the causal agent of white mould, is a necrotrophic fungal pathogen responsible for extensive crop loss. Current control options rely heavily on the application of chemical fungicides that are becoming less effective and may lead to the development of fungal resistance. In the current study, we used a foliar spray application of boron to protect Brassica napus (canola) from S. sclerotiorum infection using whole plant infection assays. Application of boron to aerial surfaces of the canola plant reduced the number of S. sclerotiorum forming lesions by 87% compared to an untreated control. We used dual RNA sequencing to profile the effect of boron on both the host plant and fungal pathogen during the infection process. Differential gene expression analysis and gene ontology term enrichment further revealed the mode of action of a foliar boron spray at the mRNA level. A single foliar application of boron primed the plant defense response through the induction of genes associated with systemic acquired resistance while an application of boron followed by S. sclerotiorum infection induced genes associated with defense-response-related cellular signalling cascades. Additionally, in S. sclerotiorum inoculated on boron-treated B. napus, we uncovered gene activity in response to salicylic acid breakdown, consistent with salicylic-acid-dependent systemic acquired resistance induction within the host plant. Taken together, this study demonstrates that a foliar application of boron results in priming of the B. napus plant defense response, likely through systemic acquired resistance, thereby contributing to increased tolerance to S. sclerotiorum infection.