Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Two contrasting genotypes of Theobroma cacao differ in their transcriptional response and in accumulation of reactive oxygen species in response to treatment with salicylic acid

ABSTRACT: In order to identify variety-specific differences in defense response induction, Scavina 6 (Sca6) and Imperial College Selection 1 (ICS1) cacao plantlets were treated with 2mM SA or water as control and leaves of three developmental stages (A,C, E) were collected. This process was repeated 3 times yielding 32 samples, (Stage E ICS1 and Stage A Sca6 were collected only twice.) Averaging across developmental stages within each genotype, ICS1 had 436 up- and 601 down-regulated genes and Sca6 had 490 up- and 447 down-regulated genes (Benjamini-Hochberg p-value < 0.05). Analysis of gene annotations and Gene Ontology terms revealed that more PR genes and genes associated with defense response were up-regulated in ICS1, the more pathogen-susceptible genotype, while Sca6 had more genes associated with photosynthesis and energy generation. Same 17k unigene array as described in Mejía LC, Herre EA, Sparks JP, Winter K, García MN, Van Bael SA, Stitt J, Shi Z, Zhang Y, Guiltinan MJ, Maximova SN. 2014. Pervasive effects of a dominant foliar endophytic fungus on host genetic and phenotypic expression in a tropical tree. Frontiers in Microbiology 5.

ORGANISM(S): Theobroma cacao

SUBMITTER: Mark Guiltinan

PROVIDER: E-GEOD-64022 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Salicylic acid (SA) is a critical molecule mediating plant innate immunity with an important role limiting the growth and reproduction of the virulent powdery mildew (PM) Golovinomyces orontii on Arabidopsis thaliana. To investigate this later phase of the PM interaction, and the role played by SA, we performed replicated global expression profiling for wild type and SA biosynthetic mutant ics1 Arabidopsis from 0 to 7 days post infection. We found that ICS1-impacted genes comprise 3.8% of profiled genes with known molecular markers of Arabidopsis defense ranked very highly by the multivariate empirical Bayes statistic (T2 statistic ((Tai and Speed, 2006)). Functional analyses of T2-selected genes identified statistically significant PM-impacted processes including photosynthesis, cell wall modification, and alkaloid metabolism that are ICS1-independent. ICS1-impacted processes include redox, vacuolar transport/secretion, and signaling. Our data also supports a role for ICS1 (SA) in iron and calcium homeostasis and identifies components of SA crosstalk with other phytohormones. Through our analysis, 39 novel PMâimpacted transcriptional regulators were identified. Insertion mutants in one of these regulators, PUX2, results in significantly reduced reproduction of the powdery mildew in a cell death independent manner. Though little is known about PUX2, PUX1 acts as a negative regulator of Arabidopsis CDC48 (Rancour et al., 2004; Park et al., 2007), an essential AAA-ATPase chaperone that mediates diverse cellular activities including homotypic fusion of ER and Golgi membranes, ER-associated protein degradation, cell cycle progression, and apoptosis. Future work will elucidate the functional role of the novel regulator PUX2 in PM resistance. Experiment Overall Design: Arabidopsis whole leaves were harvested at 6h, 1 day, 3 days, 5 days and 7 days after Golovinomyces orontii infection for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. Gene expression profiles were obtained for wild type Columbia-0 and enhanced disease susceptibility mutant eds16-1, a null isochorismate synthase 1 (At1g74710) mutant. In parallel, uninfected samples were collected at 0 hr and 7days from wild type and mutant plants. The experiment includes 4 biological replicates.

Project description:Lipoprotein lipase (LPL) is responsible for the intravascular catabolism of triglyceride-rich lipoproteins and plays a central role in whole-body energy balance and lipid homeostasis. As such, LPL is subject to tissue-specific regulation in different physiological conditions, but the mechanisms of this regulation remain incompletely characterized. Previous work revealed that LPL comprises a set of proteoforms with different isoelectric points, but their regulation and functional significance have not been studied thus far. Here we studied the distribution of LPL proteoforms in different rat tissues and their regulation under physiological conditions. First, analysis by two-dimensional electrophoresis and Western blot showed different patterns of LPL proteoforms (i.e., different pI or relative abundance of LPL proteoforms) in different rat tissues under basal conditions, which could be related to the tissue-specific regulation of the enzyme. Next, the comparison of LPL proteoforms from heart and brown adipose tissue between adults and 15-day-old rat pups, two conditions with minimal regulation of LPL in these tissues, yielded virtually the same tissue-specific patterns of LPL proteoforms. In contrast, the pronounced down-regulation of LPL activity observed in white adipose tissue during fasting is accompanied by a prominent reconfiguration of the LPL proteoform pattern. Furthermore, refeeding reverts this down-regulation of LPL activity and restores the pattern of LPL proteoforms in this tissue. Importantly, this reversible proteoform-specific regulation during fasting and refeeding indicates that LPL proteoforms are functionally diverse. Further investigation of potential differences in the functional properties of LPL proteoforms showed that all proteoforms exhibit lipolytic activity and have similar heparin-binding affinity, although other functional aspects remain to be investigated. Overall, this study demonstrates the ubiquity, differential distribution and specific regulation of LPL proteoforms in rat tissues and underscores the need to consider the existence of LPL proteoforms for a complete understanding of LPL regulation under physiological conditions.

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Two contrasting genotypes of Theobroma cacao differ in their transcriptional response and in accumulation of reactive oxygen species in response to treatment with salicylic acid

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