Dataset Information

Effect of ?-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of Lasiodiplodia theobromae Isolated from Grapevine.

ABSTRACT: The effect of ?-aminobutyric acid (GABA) on the metabolome of two strains of Lasiodiplodia theobromae isolated from grapevine that hold a different degree of virulence to the host plant (LA-SOL3 (more virulent), LA-SV1 (less virulent)) was investigated. The culture filtrates and crude extracts from the two strains grown in the presence and absence of 10 mM of GABA were tested for phytotoxicity on tomato plant cuttings and leaves, respectively. Considering the opportunistic nature of this fungus for humans, crude extracts were also tested for cytotoxicity on mammalian cell lines. We found that culture filtrates and crude extracts have a decreased toxicity in the presence of GABA. Metabolomic analysis, conducted on both strains at both growth conditions, revealed the production of several compounds, such as indole-3-carboxylic acid (ICA, which is the main compound produced by L. theobromae), 3-indolecarboxyaldehyde, (3R,4S)-botryodiplodin, (R)-mellein. Finally, data demonstrate that GABA both induces a decrease in the amount of ICA, and a diversification of the metabolites produced by L. theobromae.

SUBMITTER: Salvatore MM

PROVIDER: S-EPMC7503889 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Effect of γ-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of <i>Lasiodiplodia theobromae</i> Isolated from Grapevine.

Salvatore Maria Michela MM Félix Carina C Lima Fernanda F Ferreira Vanessa V Duarte Ana Sofia AS Salvatore Francesco F Alves Artur A Esteves Ana Cristina AC Andolfi Anna A

Molecules (Basel, Switzerland) 20200823 17

The effect of γ-aminobutyric acid (GABA) on the metabolome of two strains of <i>Lasiodiplodia theobromae</i> isolated from grapevine that hold a different degree of virulence to the host plant (LA-SOL3 (more virulent), LA-SV1 (less virulent)) was investigated. The culture filtrates and crude extracts from the two strains grown in the presence and absence of 10 mM of GABA were tested for phytotoxicity on tomato plant cuttings and leaves, respectively. Considering the opportunistic nature of this ...[more]

PMID: 32842542

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Project description:BackgroundLasiodiplodia theobromae is a fungus of the Botryosphaeriaceae that causes grapevine vascular disease, especially in regions with hot climates. Fungi in this group often remain latent within their host and become virulent under abiotic stress. Transcriptional regulation analysis of L. theobromae exposed to heat stress (HS) was first carried out in vitro in the presence of grapevine wood (GW) to identify potential pathogenicity genes that were later evaluated for in planta expression.ResultsA total of 19,860 de novo assembled transcripts were obtained, forty-nine per cent of which showed homology to the Botryosphaeriaceae fungi, Neofusicoccum parvum or Macrophomina phaseolina. Three hundred ninety-nine have homology with genes involved in pathogenic processes and several belonged to expanded gene families in others fungal grapevine vascular pathogens. Gene expression analysis showed changes in fungal metabolism of phenolic compounds; where genes encoding for enzymes, with the ability to degrade salicylic acid (SA) and plant phenylpropanoid precursors, were up-regulated during in vitro HS response, in the presence of GW. These results suggest that the fungal L-tyrosine catabolism pathway could help the fungus to remove phenylpropanoid precursors thereby evading the host defense response. The in planta up-regulation of salicylate hydroxylase, intradiol ring cleavage dioxygenase and fumarylacetoacetase encoding genes, further supported this hypothesis. Those genes were even more up-regulated in HS-stressed plants, suggesting that fungus takes advantage of the increased phenylpropanoid precursors produced under stress. Pectate lyase was up-regulated while a putative amylase was down-regulated in planta, this could be associated with an intercellular growth strategy during the first stages of colonization.ConclusionsL. theobromae transcriptome was established and validated. Its usefulness was demonstrated through the identification of genes expressed during the infection process. Our results support the hypothesis that heat stress facilitates fungal colonization, because of the fungus ability to use the phenylpropanoid precursors and SA, both compounds known to control host defense.

Project description:Gummosis is 1 of the most common and destructive diseases threatening global peach (Prunus persica) production. Our previous studies have revealed that ethylene and methyl jasmonate enhance peach susceptibility to Lasiodiplodia theobromae, a virulent pathogen inducing gummosis; however, the underlying molecular mechanisms remain obscure. Here, 2 ethylene response factors (ERFs), PpERF98 and PpERF1, were identified as negative regulators in peach response to L. theobromae infection. Expression of 2 putative paralogs, PpERF98-1/2, was dramatically induced by ethylene and L. theobromae treatments and accumulated highly in the gummosis-sensitive cultivar. Silencing of PpERF98-1/2 increased salicylic acid (SA) content and pathogenesis-related genes PpPR1 and PpPR2 transcripts, conferring peach resistance to L. theobromae, whereas peach and tomato (Solanum lycopersicum) plants overexpressing either of PpERF98-1/2 showed opposite changes. Also, jasmonic acid markedly accumulated in PpERF98-1/2-silenced plants, but reduction in PpPR3, PpPR4, and PpCHI (Chitinase) transcripts indicated a blocked signaling pathway. PpERF98-1 and 2 were further demonstrated to directly bind the promoters of 2 putative paralogous PpERF1 genes and to activate the ERF branch of the jasmonate/ethylene signaling pathway, thus attenuating SA-dependent defenses. The lesion phenotypes of peach seedlings overexpressing PpERF1-1/2 and PpERF98-1/2 were similar. Furthermore, PpERF98-1/2 formed homodimers/heterodimers and interacted with the 2 PpERF1 proteins to amplify the jasmonate/ethylene signaling pathway, as larger lesions were observed in peach plants cooverexpressing PpERF98 with PpERF1 relative to individual PpERF98 overexpression. Overall, our work deciphers an important regulatory network of ethylene-mediated peach susceptibility to L. theobromae based on a PpERF98-PpERF1 transcriptional cascade, which could be utilized as a potential target for genetic engineering to augment protection against L. theobromae-mediated diseases in crops and trees.

Project description:High temperatures associated with a fluctuating climate profoundly accelerate the occurrence of a myriad of plant diseases around the world. A comprehensive insight into how plants respond to pathogenic microorganisms under high-temperature stress is required for plant disease management, whereas the underlying mechanisms behind temperature-mediated plant immunity and pathogen pathogenicity are still unclear. Here, we evaluated the effect of high temperature on the development of grapevine canker disease and quantified the contribution of temperature variation to the gene transcription reprogramming of grapevine and its pathogenic agent Lasiodiplodia theobromae using a dual RNA-seq approach. The results showed that both grapevine and the pathogen displayed altered transcriptomes under different temperatures, and even the transcription of a plethora of genes from the two organisms responded in different directions and magnitudes. The transcription variability that arose due to temperature oscillation allowed us to identify a total of 26 grapevine gene modules and 17 fungal gene modules that were correlated with more than one gene module of the partner organism, which revealed an extensive web of plant-pathogen gene reprogramming during infection. More importantly, we identified a set of temperature-responsive genes that were transcriptionally orchestrated within the given gene modules. These genes are predicted to be involved in multiple cellular processes including protein folding, stress response regulation, and carbohydrate and peptide metabolisms in grapevine and porphyrin- and pteridine-containing compound metabolisms in L. theobromae, implying that in response to temperature oscillation, a complex web of signaling pathways in two organism cells is activated during infection. This study describes a co-transcription network of grapevine and L. theobromae in the context of considering temperature variation, which provides novel insights into deciphering the molecular mechanisms underlying temperature-modulated disease development.

Project description:Purpose: Lasiodiplodia theobromae is a pathogenic fungus causing Botryosphaeria dieback in grapevine. Heat and drought stress was suggested to promote the disease incidence in recent years, therefore we decided to evaluate the effect of heat on in vitro experiment, adding grapevine wood to simulate the environment in the trunk during in field infection. Methods: Fungal mRNA was extracted from mycellium collected after 49 hours of growth without stress in the presence of GW (GW_noHS) or in its absence (VS_noHS). Some mRNA were extracted form mycellium collected 10 min after stress (1 hour at 42 C) both in the presence of GW (GW_HS) or in its absence (VS_HS). The cDNA and sequenciation was done according with Illumina HiSeq 2500 recomendations by triplicate. Paired end reads (100bp) were used for de novo assembly through Trinity. Transdecoder was used to predict ORFs. The transcripts functional annotatio was done through BlastX and Blast2GO. Bowtie2 was used to map the reads for each sample to the previously assembled transcritome, a homemade Perl script was used to count the mapped reads and edgeR was employed for differential expression and statistical analysis. Results: Seventy million reads were asembled to produce 19,860 transcripts, from those 15,981 were predicted to codify for ORFs. Thirty-five % of transcripts did not have any homologue in NCBI NR database, most of those also did not were predicted to codify for ORFs. With the remainning 38 % of transcripts the functional annotation was obtained. Around 90% of reads for each sample were mapped to the de novo assembled transcriptome. Around 2339 genes were differentially expressed when comparing GW_HS vs GW_noHS or vs VS_noHS and GW_noHS vs VS_noHS and VS_HS vs VS_noHS. Hierarchical clustering with functional enrichment were used to predict metabolic pathways with differential regulation. In silico prediction were then verified with RT-qPCR showing linear correlation (r2=0.87) in in vitro experiment. Then trough in planta fungal gene expression helps to verify that phenolic metabolism is important for colonization in grapevine. Conclusion: A model for fungal response to HS in the presence of GW was proposed. This correspond to the first work to identify nucleotide sequences, measure gene expression and propose pathogenicity factors important for disease establishment in grapevine.

Dataset Information

Effect of ?-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of Lasiodiplodia theobromae Isolated from Grapevine.

Publications

Effect of γ-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of <i>Lasiodiplodia theobromae</i> Isolated from Grapevine.

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