Project description:We used Arabidopsis full-genome microarrays to characterize plant transcript accumulations at different stages of infection with the biotrophic oomycete downy mildew pathogen, Hyaloperonospora arabidopsidis : initiation (< 1 dpi) and maintenance of infection (> 4 dpi). In two independent experiments, cotyledons from the ecotype Wassilewskija (WS) were inoculated with water, or with Hyaloperonospora arabidopsidis to establish a compatible interaction. Affymetrix ATH1 microarrays were used to profile Arabidopsis transcript accumulations at the initiation (mixed samples at 8 and 24 hours post inoculation, hpi; early stage) and maintenance (mixed samples at 4 and 6 days post inoculation; late stage) of the compatible interaction.
Project description:The biological function of the plant-microbiome system is the result of contributions from the host plant and microbiome members. The Populus root microbiome is a diverse community that has high abundance of β- and γ-Proteobacteria, both classes which include multiple plant-growth promoting representatives. To understand the contribution of individual microbiome members in a community, we studied the function of a simplified community consisting of Pseudomonas and Burkholderia bacterial strains isolated from Populus hosts and inoculated on axenic Populus cutting in controlled laboratory conditions. Both strains increased lateral root formation and root hair production in Arabidopsis plate assays and are predicted to encode for different functions related to growth and plant growth promotion in Populus hosts. Inoculation individually, with either bacterial isolate, increased root growth relative to uninoculated controls, and while root area was increased in mixed inoculation, the interaction term was insignificant indicating additive effects of root phenotype. Complementary data including photosynthetic efficiency, whole-transcriptome gene expression and GC-MS metabolite expression data in individual and mixed inoculated treatments indicate that the effects of these bacterial strains are unique and additive. These results suggest that the function of a microbiome community may be predicted from the additive functions of the individual members.
Project description:Using pine wood nematode resistant Pinus massoniana clones as materials, after inoculation with pine wood nematode, needles from 5 locations of the same plant were collected and mixed as a biological replicate at 0d,3d and 10d, and a total of two biological replicates were performed for proteomics analysis based on TMT tags.
Project description:Plants of the resistant Pisum sativum subsp. syriacum accession P665 and the susceptible pea cultivar Messire were inoculated with M. pinodes.The experiment was conducted in three replicates. 16, 24 and 48 hours after inoculation RNA was isolated from leaves of infected plants and transcribed into cDNA. For each time point and replicate, Cy-labelled cDNA samples from resistant and susceptible plants were mixed and hybridized to Mt16kOLI1Plus microarray
Project description:Three plants of two species in Brassica genus, i.e. B. rapa cv. "Purple To White Globe" and B. napus cv. "SariGol", highly susceptible and tolerant hosts of Cauliflower mosaic virus (CaMV), respectively, were mechanically inoculated with CaMV strain NY8153.Phosphate buffer was used as control in mock-inoculated plants. RNAs, in three biological repeats, were extracted 20 days post-inoculation (dpi) collecting ten leaves from mock-inoculated and virus-infected plants using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. Five µg of total RNA were resolved in 15% SDS-PAGE gel. Total RNA pattern was visualized upon Ethidium Bromide staining and sRNAs in the size range of 20-30 nt were eluted from gel. sRNAs replicates were quantified and mixed in equal molarity. Libraries were obtained using NEXTflex Small RNA-Seq Kit v3 (Bio Scientific) according to manufacturer’s instructions. Libraries were eluted from a 6% non-denaturing PAGE. The quality of libraries was checked by BioPharma QC, and then, submitted for sequencing with a HiSeq 2500 Illumina platform.
Project description:To understand the plant-pathogen interaction comprehensively, it is valuable to monitor the gene expression profiles of both interacting organisms simultaneously in the same infected plant tissue. Using RNA-Seq, we analyzed the mixed transcriptome of rice and blast fungus in infected leaves at 24 hours post-inoculation. We demonstrated that our method detected the gene expression of both the host plant and pathogen simultaneously in the same infected leaf blades in natural infection conditions without any artificial treatments. Using compatible (Ina86-137) and incompatible (P91-15B) fungal strains as pathogens, we revealed the differential expression profiles of the compatible and incompatible interactions and observed that the responsive gene expression was more drastic in the incompatible interaction. Our mixed transcriptome analysis is useful for the simultaneous elucidation of the tactics of host plant defense and pathogen attack.
Project description:In the study, two soybean genotypes were selected to conduct high and low P, high and low Mg, and AM fungal inoculation treatments, combined with RNA-seq sequencing technique in order to investigate the physiological and molecular mechanisms of the symbiosis between soybean and AM fungi affected by P and Mg treatments. The results showed that both Mg application and AM fungal inoculation were beneficial to promote soybean growth under low P condition. And there was a synergistic effect between the Mg concentration and the P concentration in the root of HN112 under the inoculation condition. RNA-seq sequencing was carried out using the roots of P-efficient soybean HN89 under different Mg and inoculation treatments with low P condition, and the difference of gene expression profiles was analyzed between high and low Mg treatments, and different inoculation treatments. According to the analysis of GO function classification and KEGG enrichment, under high Mg condition, the metabolic pathway was mainly enriched in lipid metabolism and glucose metabolism pathway under the inoculation treatment compared with the non-inoculation treatment, which regulated carbon metabolism pathway. Under the low Mg condition, the metabolic pathway was mainly enriched in the photosynthesis- antenna protein pathway to regulate the photosynthesis pathway under the inoculation treatment compared with the non-inoculation treatment. At the same time, the inoculation treatment significantly increased soybean root starch concentration under low Mg condition, compared with the non-inoculation treatment, suggesting that the significant up-regulation of a large number of photosynthesis related genes might be related to the significant increase of starch concentration at this treatment.