DataSheet_1_Deciphering the genetic architecture of resistance to Corynespora cassiicola in soybean (Glycine max L.) by integrating genome-wide association mapping and RNA-Seq analysis.zip

Target spot caused by Corynespora cassiicola is a problematic disease in tropical and subtropical soybean (Glycine max) growing regions. Although resistant soybean genotypes have been identified, the genetic mechanisms underlying target spot resistance has not yet been studied. To address this knowledge gap, this is the first genome-wide association study (GWAS) conducted using the SoySNP50K array on a panel of 246 soybean accessions, aiming to unravel the genetic architecture of resistance. The results revealed significant associations of 14 and 33 loci with resistance to LIM01 and SSTA C. cassiicola isolates, respectively, with six loci demonstrating consistent associations across both isolates. To identify potential candidate genes within GWAS-identified loci, dynamic transcriptome profiling was conducted through RNA-Seq analysis. The analysis involved comparing gene expression patterns between resistant and susceptible genotypes, utilizing leaf tissue collected at different time points after inoculation. Integrating results of GWAS and RNA-Seq analyses identified 238 differentially expressed genes within a 200 kb region encompassing significant quantitative trait loci (QTLs) for disease severity ratings. These genes were involved in defense response to pathogen, innate immune response, chitinase activity, histone H3-K9 methylation, salicylic acid mediated signaling pathway, kinase activity, and biosynthesis of flavonoid, jasmonic acid, phenylpropanoid, and wax. In addition, when combining results from this study with previous GWAS research, 11 colocalized regions associated with disease resistance were identified for biotic and abiotic stress. This finding provides valuable insight into the genetic resources that can be harnessed for future breeding programs aiming to enhance soybean resistance against target spot and other diseases simultaneously.</p

CB2-R gene targeting modifies behavior.

<p><i>A</i>, Behavioral effects of CB2 intracerebral gene targeting by antisense oligonucleotide microinjected into the mouse brain and performance of mice in plus-maze test was assessed before and after 3 days of twice daily microinjection. AS1 and AS2 were before and after CB2 antisense oligo microinjection. V1 and V2 are controls. <i>B</i>, performance in plus-maze test following CMS or mice exposed prenatally to capsaicin and the effect of JWH015 (20 mg/kg).</p

Brain CB2-Rs: Immunohistochemistry in mouse and rat brain.

<p><i>A</i>, CB2-IR in apical dendrites and cell bodies of pyramidal neurons of rat cerebral cortex. <i>B</i>, CB2-IR in mouse cerebral cortex. <i>C</i>, CB2-IR in rat corpus callosum and <i>D</i>, CB2-IR in mouse hippocampal allocortex and some interneurons in the striatum oriens and stratum radiatum.</p

Brain CB2-Rs: Immunoractivity (IR) and pre-adsorption with immunizing peptide.

<p><i>A</i>, CB2-IR in the left panel and lack of CB2-IR in the right panel, <i>B</i>, when the CB2 antibody was pre-adsorped with the peptide.</p

CB2-R- activation and blockade on anhedonia induced by chronic mild stress (CMS).

<p><i>A</i>, Weekly sucrose consumption in stress and control mice. <i>B</i>, Effect of JWH015 (20 mg/kg) on mouse weekly sucrose consumption test. C, Effect of AM630 (1 and 3 mg/kg) on mouse weekly sucrose consumption test.</p

Allelic and genotype distribution of R63Q polymorphism in the <i>CB2 gene</i>.

<p>Comparisons were made between patients with major depression and the healthy controls. Significant differences are observed in allelic frequency and genotype distribution in the R recessive model. OR is odd ratio.</p

Presence of <i>CB2</i> gene in the brain.

<p>A, Relative brain expression of <i>CB2</i> gene in C57BL/6J and BALBc strains subjected to stress. <i>B</i>, Relative <i>CB2</i> gene expression levels in the striatum, midbrain, and hippocampus of C57Bl/6J mice. <i>C</i>, Mouse whole brain relative <i>CB2</i> gene expression levels following chronic treatment with heroin and cocaine. <i>D</i>, relative <i>CB2</i> gene expression levels in striatum and midbrain of mice that developed alcohol preference. <i>CB2</i> gene expression was relative to the standard laboratory brain obtained from C57BL/6J that was set to 1.0. The positive control was from the spleen and no cDNA in TaqMan PCR reaction served as negative controls.</p

Brain CB2-Rs: Immunoblots, genotyping and <i>in-situ</i> hybridization.

<p><i>A</i>, <i>In-situ</i> hybridization indicating <i>CB2</i> gene is expressed in the cerebellum of wild type and not in the cerebellum of the CB2-R deficient mice and also in sense controls in the wild type mice. <i>B</i>, RFLP genotyping discrimination on agarose gel for <i>CB2</i> Q63R polymorphism in depressed subjects (Ba) and, Resequences of <i>CB2</i> Q63R polymorphism (Bb). <i>C</i>, Western blotting of CB2-Rs in CMS and control mice (left panel) and in right panel in mice exposed to 4 mg/kg capsaicin in utero.</p

Behavioral effects of CB2-R activation and blockade.

<p><i>A</i>, Mouse spontaneous locomotor activity following acute treatment with CB2 agonist JWH015 (1–20 mg/kg), in mouse strain, C57Bl/6 (a and b); BALBc, (c and d) and DBA/2 (e and f). <i>B</i>, Effect of JWH015 in C57Bl/6 mice in the two compartment black and white box, showing time spent in the black and white chamber. <i>C</i>, Acute effects of SR144528 – a CB2-R antagonist on DBA/2 mouse spontaneous locomotor activity and stereotype behavior. <i>D</i>, Acute effects of SR144528, in DBA/2 male and female mice in the two chamber black and white test box, showing time spent in the black and white chamber.</p

Subcellular localization of rat hippocampal CB2-Rs.

<p><i>A</i>, a CB2-IR dendrite [CB2(+) D] receiving multiple synaptic contacts from axon terminals lacking CB2-R immunolabeling [CB2(−) AT]. B, a CB2-IR dendrite [CB2(+) D] was contacted by a non-immunoreactive axon terminal [CB2(−) AT]. Scale bar represents 0.3 µm.</p