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Identification of quantitative trait loci controlling gene expression during the innate immunity response of soybean

ABSTRACT: Microbe associated molecular pattern (MAMP)-responsive genes were identified in leaves of four different genotypes. The genotypes used in this study were: LD0-2817P; LDX01-1-65; Ripley and EF59. Leaves from three-weeks old plants were used in this study We used microarray to identified the MAMP-responsive genes in leaves of four soybean genotypes trated with 1 uM flg22 and 50 ug/ml chitin for 30 minutes Trifoliolate leaves from three-week-old plants were detached and then vacuumed infiltrated with ddiH2O for 2 min. Water-infiltrated trifoliolate leaves from five different plants of each genotype were pooled and cut into approximately 1 cm2 slices. Equal amount of leaf slices (~30 slices) were transferred into two different Petri dishes and then floated overnight on autoclaved ddiH2O. Water was removed from both Petri dishes and was replaced with 5 ml of MAMP solution [1 mM flagellin 22 (flg22) and 50 mg of crab shell chitin (called in this study as chitin)], or 5 ml of mock solution [autoclaved ddiH2O plus equivalent amount of dimethyl sulfoxide (DMSO). DMSO was included since it was contained in the solution used to dissolve the flg22 peptide]. After a 30 min treatment, mock- and MAMP-treated leaf slices were harvested into different tubes and immediately frozen in liquid nitrogen. Samples were stored at -80 until use. All procedures described above were performed under dark conditions.

ORGANISM(S): Glycine max

SUBMITTER: Oswaldo Valdes-Lopez

PROVIDER: E-GEOD-32642 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Identification of quantitative trait loci controlling gene expression during the innate immunity response of soybean.

Valdés-López Oswaldo O Thibivilliers Sandra S Qiu Jing J Xu Wayne Wenzhong WW Nguyen Tran H N TH Libault Marc M Le Brandon H BH Goldberg Robert B RB Hill Curtis B CB Hartman Glen L GL Diers Brian B Stacey Gary G

Plant physiology 20110930 4

Microbe-associated molecular pattern-triggered immunity (MTI) is an important component of the plant innate immunity response to invading pathogens. However, most of our knowledge of MTI comes from studies of model systems with relatively little work done with crop plants. In this work, we report on variation in both the microbe-associated molecular pattern-triggered oxidative burst and gene expression across four soybean (Glycine max) genotypes. Variation in MTI correlated with the level of pat ...[more]

PMID: 21963820

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Project description:Despite their importance, plant MAP kinase targets are still poorly elucidated. Here, the specific in vivo interaction of an ethylene response factor (ERF104) with the Arabidopsis MAP kinase, MPK6, is shown by fluorescence resonance energy transfer. The interaction, which is lost within minutes after treatment with the flagellin-derived flg22 peptide, is dependent on both MPK6 kinase activity and rapid ethylene signaling initiated downstream of MPK6 activation. ERF104 is an MPK6 substrate and phosphorylation site mutations affected its stability. ERF104 activates promoters with GCC elements. This was evident from microarray data of overexpressing transgenic plants, where promoters of up regulated genes contain GCC motifs and chromatin immunoprecipitation showing ERF104 association with PDF1.2 promoter. The ERF104 overexpressor did not affect biotrophic bacteria proliferation but was more susceptible to necrotrophic Botrytis cinerea. Microarray performed with erf104 or mpk6 revealed only a limited number of flg22-induced genes that require these elements - possibly as a; result of functional redundancies. Thus, ERF104 phosphorylation by MPK6, in concert with ethylene signaling induced by pathogen-derived molecules, modulates defense in Arabidopsis. Experiment Overall Design: Leaves of six week old Col-0, erf104, mpk6 and 35S::ERF104 plants were infiltrated with 1ÂµM Flg22 or water and harvested four hours later. Total RNA was isolated and processed according to the Affymetrix protocol for biotin-labelled cRNA and hybridized to the Affymetrix ATH1 chip. The data Experiment Overall Design: was analyzed with Genespring GX 7.3.1 software (Agilent) with the following parameters: Filter on Flags for present or marginal in 50% of all considered experiments, Filter for reliable differentially expressed genes based on volcano plot (one way ANOVA p-value <0.05 and >3-fold change in expression). For the flg22 experiments, analysis for each genotype was separately performed and a composite list of flg22-regulated genes compiled â with the aim of including genes that may be differentially regulated in the genotype. Global expression profile was visualized by k-means clustering and condition tree (Genespring).

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Identification of quantitative trait loci controlling gene expression during the innate immunity response of soybean

Publications

Identification of quantitative trait loci controlling gene expression during the innate immunity response of soybean.

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