Dataset Information

A novel protein extraction method for proteomics of the trunk disease fungus Lasiodiplodia theobromae (Botryosphaeriaceae)

ABSTRACT: Fungal proteomics is a developing field that requires renewed interest and attention from the scientific community in many aspects. One of the most compelling objectives of fungal biology is to find out how fungal pathogens colonize a host, in order to find new ways of impeding this colonization, and proteomics is an important and useful tool to this end. However, fungi are also taxonomically interesting and are important biochemical model organisms for the study of many cellular processes, such as cytoskeletal regulation. Additionally, fungi are sources of secondary metabolites, many of which serve as medications for humans. Nevertheless, much work remains in developing systems-biology approaches to understanding fungal gene expression and secondary metabolism. Current fungal proteomics approaches involve 2D SDS-PAGE and extensive, complex, protein extraction methodologies. In this work, an application of a modified Folch extraction to protein extraction was used to perform de novo peptide sequencing of the proteome from the plant and human pathogen Lasiodiplodia theobromae, which greatly streamlined and simplified the analysis process. Using a metaproteomics bioinformatics approach, many novel proteins for L. theobromae were identified and targeted for further biochemical characterization and annotation efforts.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Lasiodiplodia Theobromae

TISSUE(S): Mycelium

DISEASE(S): Pneumonia,Keratitis,Wounds And Injuries,Tinea Unguium

SUBMITTER: Carla Uranga

LAB HEAD: Majid Ghassemian, PhD

PROVIDER: PXD005283 | Pride | 2017-04-05

REPOSITORIES: Pride

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Publications

Fatty acid esters produced by Lasiodiplodia theobromae function as growth regulators in tobacco seedlings.

Uranga Carla C CC Beld Joris J Mrse Anthony A Córdova-Guerrero Iván I Burkart Michael D MD Hernández-Martínez Rufina R

Biochemical and biophysical research communications 20160227 2

The Botryosphaeriaceae are a family of trunk disease fungi that cause dieback and death of various plant hosts. This work sought to characterize fatty acid derivatives in a highly virulent member of this family, Lasiodiplodia theobromae. Nuclear magnetic resonance and gas chromatography-mass spectrometry of an isolated compound revealed (Z, Z)-9,12-ethyl octadecadienoate, (trivial name ethyl linoleate), as one of the most abundant fatty acid esters produced by L. theobromae. A variety of natural ...[more]

PMID: 26926564

Publication: 1/2

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Project description:Inoculation of endophyte-free (E-) Theobroma cacao leaves with Colletotrichum tropicale (E+), the dominant foliar fungal endophyte in healthy T. cacao, induced significant changes in the expression of hundreds of host genes. Further, E+ leaves exhibit enhanced pathogen resistance, increased lignin and cellulose content, reduced maximum rates of photosynthesis (Amax), and enrichment of nitrogen-15 and carbon-13 isotopes that all correspond to the changes in expression of specific functional genes in related pathways. Moreover, a cacao gene highly up-regulated in E+ leaves increases pathogen resistance apart from any direct endophyte effects. Thus, benefits of increased pathogen resistance in E+ plants are partially due to enhanced induction of intrinsic host defense pathways, and potential costs include reduced photosynthetic capacity and endophyte metabolism of host tissues. Similar effects are likely to be properties of most plant-endophyte interactions, suggesting general relevance to the design and interpretation of studies of genetic and phenotypic expression in plants. The objective of this experiment was to identify Theobroma cacao genes that are differentially expressed between leaves inoculated with fungal endophyte Colletotrichum tropicale (E+ leaves) and control un-inoculated leaves (E- leaves) 3 days post endophyte inoculation. The experiment was conducted in a Percival growth chamber (model I35LL, 115 volts, 1/4 Hp, series: 8503122.16, Percival Scientific, Inc., Perry IA) with 12/12 h light/dark photoperiod and temperatures of 30M-BM-:C and 26M-BM-:C respectively. Inoculation was done by aspersion of endophyte spores (2X10^6 spore/ml) to a group of T. cacao seedlings and a second group of seedlings were maintained as control un-inoculated (E- leaves). Then three biological replicates (each one consisting of one leaf from different plants) per treatment E+ and four leaves per treatment E- leaves) were collected and processed for a two color oligo microarray analysis.

Dataset Information

A novel protein extraction method for proteomics of the trunk disease fungus Lasiodiplodia theobromae (Botryosphaeriaceae)

Publications

Fatty acid esters produced by Lasiodiplodia theobromae function as growth regulators in tobacco seedlings.

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