LOX1 and PLP1 dependent transcriptional reprogramming is essential for injury induced conidiophore development in a filamentous fungus
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ABSTRACT:
PROVIDER: PRJNA985641 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR24975260_1.fastq.gz | Fastqsanger.gz | |||
| SRR24975260_2.fastq.gz | Fastqsanger.gz | |||
| SRR24975261_1.fastq.gz | Fastqsanger.gz | |||
| SRR24975261_2.fastq.gz | Fastqsanger.gz | |||
| SRR24975262_1.fastq.gz | Fastqsanger.gz |
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Project description:We need to learn more about the components of the fungal damage response that are similar and well-studied in plants and animals, such as the role of oxidized lipids in mechanical stress. Using a genetic approach, mass spectrometry, and phenotypic analysis, we describe the functional role of a patatin-like phospholipase (plp1) and a unique lipoxygenase (lox1) with biosynthetic function in the production of oxidized PUFA derivatives by mechanical injury in the model fungus Trichoderma atroviride. lox1 and plp1 are co-expressed and regulated by damage signaling and sensing components. Phenotypic analysis revealed an essential defect in the emergence of aerial hyphae in the lox1 and plp1 null mutant strains, blocking injury-induced conidiation. In addition, functional loss analysis demonstrated that both genes are essential for producing the wound oxylipin 13-HODE (13-hydroxy octadecadienoic acid) and the transcriptional reprogramming required for conidiation. T. atroviride requires LOX1 and PLP1 to induce at early stages of the response transcription factors involved in asexual development like a BrlA homolog, Hox2, and Azf1, and at a later stage to activate lipid metabolism and structural proteins involved in aerial mycelium emergence. Our study emphasizes how the co-function of lox1 and plp1, during the response to wounding, regulate the molecular and physiological processes that lead to reproductive aerial mycelium development to warrant survival through conidiation.
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Project description:The ability to respond to injury is essential for the survival of an organism and involves analogous mechanisms in animals and plants. Such mechanisms integrate coordinated genetic and metabolic reprogramming events requiring fine tuned regulation by small RNAs for adequate healing of the wounded area. We previously reported that in response to injury, the filamentous fungus Trichoderma atroviride uses molecular mechanisms very similar to those of plants and animals. These processes lead to the formation of new hyphae and the development of asexual reproduction structures (conidiophores). Here we show that regulation by sRNAs of nitrogen metabolic processes and the cellular redox balance is necessary to carry out the correct response to injury. We further discover microRNA-like RNAs (milRNAs) related with the development of conidiophores. Finally, using a loss-of-function and transcriptomic approach, we demonstrate that a milRNA plays a central role in growth and hyphal regeneration in the filamentous fungus T. atroviride, which regulates signaling and cell proliferation at the transcriptional level. Although many classes of small RNAs have been profiled in fungi, and showed to be important for contending with different types of stress, this is the first report showing a physiological function of a milRNA in fungi.
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Project description:The PFK-2/ FBPase-2 are important regulatory enzymes in the glycolytic and are present in all major eukaryotic taxa. However, unlike the well-studied animal and plant PFK-2/ FBPase-2, their functions in filamentous fungi remain little investigated. As a typical filamentous fungus and industrial microorganisms, M. thermophila was choose to investigate the functions of this gene. At the same time, the application of regulating this gene in improving metabolites production was also studied.
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Project description:The ability to respond to injury is a biological process conserved across kingdoms, which in some cases results in regeneration of an organism’s lost part or structure. Due to their immobility, multicellular fungi are prey to a variety of predators and are therefore constantly exposed to mechanical damage. Nevertheless, our current knowledge of how fungi respond to injury is scarce. Activation of injury responses and regeneration relies on the detection of danger or alarm signals known as Danger- or Damage-Associated Molecular Patterns. We show that Ca2+ influxes and extracellular ATP, two such signals, together with Mitogen-Activated Protein Kinases, play a key role in the control of hyphal regeneration in the filamentous fungus Trichoderma atroviride. We further show that these signaling cascades activate major transcriptional changes early after injury. We identified a set of regeneration genes related to cell signaling, stress responses, regulation of transcription, ribosome biogenesis/translation, replication and DNA repair. We uncovered the activation of an innate immune response, including HET domain genes, until now only known to participate in vegetative incompatibility. We show that fungal and animal regeneration share danger-signals, signaling cascades, and the activation of an immune system, challenging the current notion of the evolution of this process.
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Project description:Fungi use oxylipins (oxidized lipids) as signaling molecules to induce asexual development. These molecules play an essential role in the response to wounding, exerting a protective effect against plant pathogens, and are part of the inflammatory process in animals. However, the physiological and molecular mechanisms triggered by oxylipins that lead to asexual development in fungi are not well understood. Using a genetic approach, mass spectrometry, and phenotypic analysis, we describe the functional role of a patatin-like phospholipase (plp1) and a unique lipoxygenase (lox1) in the response to injury in the model fungus Trichoderma atroviride. lox1 and plp1 are co-expressed and regulated by damage signaling and sensing components. Phenotypic analysis revealed an essential defect in the emergence of aerial hyphae in the lox1 and plp1 null mutant strains, blocking injury-induced conidiation. In addition, functional loss analysis demonstrated that both genes are essential for wound-associated linoleic acid-derived oxylipin 13-hydroxy-9Z,11E-octadecadienoic acid (13-HODE) production and the transcriptional reprogramming required for conidiation. T. atroviride requires LOX1 and PLP1 to induce transcription factors involved in asexual development such as brlA, hox2, and azf1 homologs at the early stages of the response and at a later stage to activate lipid metabolism and the structural proteins involved in aerial mycelium emergence. Our study shows how the cooperative function of lox1 and plp1, during the response to wounding, regulates the molecular and physiological processes of damaged-sensitized cells that lead to reproductive aerial mycelium development and consequently, ensure survival through asexual reproduction.IMPORTANCEIn addition to being considered a biocontrol agent, the fungus Trichoderma atroviride is a relevant model for studying mechanisms of response to injury conserved in plants and animals that opens a new landscape in relation to regeneration and cell differentiation mechanisms. Here, we reveal the co-functionality of a lipoxygenase and a patatin-like phospholipase co-expressed in response to wounding in fungi. This pair of enzymes produces oxidized lipids that can function as signaling molecules or oxidative stress signals that, in ascomycetes, induce asexual development. Furthermore, we determined that both genes participate in the regulation of the synthesis of 13-HODE and the establishment of the physiological responses necessary for the formation of reproductive aerial mycelium ultimately leading to asexual development. Our results suggest an injury-induced pathway to produce oxylipins and uncovered physiological mechanisms regulated by LOX1 and PLP1 to induce conidiation, opening new hypotheses for the novo regeneration mechanisms of filamentous fungi.
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Project description:MicroRNAs (miRNAs) are small non-coding RNAs capable of negatively regulating gene expression. Trichoderma reesei is an industrial filamentous fungus that can secrete abundant hydrolases for cellulosic biofuels. Recently, microRNA-like RNAs (milRNAs) were discovered in several filamentous fungi rather than T. reesei. The purpose of this study was to explore the presence of milRNA in T. reesei, to characterize the differential expression of T. reesei milRNA under cellulose induction, and to reveal the target genes of milRNA involved in cellulase production. Two small RNA libraries of cellulose induction (IN) or non-induction (CON) were generated and sequenced using Solexa sequencing technology. A total of 664,463 and 529,545 unique sequences, representing 1,271 and 1,021 unique small RNAs, were obtained from the IN and CON samples, respectively. Thirteen milRNAs were finally identified in T. reesei using the hairpin structure analysis. The milRNAs profiles obtained in deep sequencing were validated by RT-qPCR assay. The miRanda program predicts a number of potential targets for T. reesei milRNAs, including several hydrolases and carbon catabolite repressor Cre1.The presence and differential expression of T. reesei milRNAs, along with their predicted targets indicate that milRNAs might play a regulatory role in cellulase induction. This work lays foundation for further functional study of fungal milRNAs and their industrial application.
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