Circadian oscillations in Trichoderma atroviride and the role of core clock components in secondary metabolism, development, and mycoparasitism against the phytopathogen Botrytis cinerea

  1. Marlene Henríquez-Urrutia
  2. Rebecca Spanner
  3. Consuelo Olivares-Yánez
  4. Aldo Seguel-Avello
  5. Rodrigo Pérez-Lara
  6. Hector Guillén-Alonso
  7. Robert Winkler
  8. Alfredo Heriberto Herrera-Estrella
  9. Paulo Canessa  Is a corresponding author
  10. Luis F Larrondo  Is a corresponding author
  1. Pontificia Universidad Católica de Chile, Chile
  2. Millennium Institute for Integrative Biology, Chile
  3. Cinvestav Unidad Irapuato, Mexico
  4. Unidad de Genómica Avanzada - Langebio, Mexico
  5. Universidad Andrés Bello, Chile

Abstract

Circadian clocks are important for an individual’s fitness, and recent studies have underlined their role in the outcome of biological interactions. However, the relevance of circadian clocks in fungal-fungal interactions remains largely unexplored. We sought to characterize a functional clock in the biocontrol agent Trichoderma atroviride to assess its importance in the mycoparasitic interaction against the phytopathogen Botrytis cinerea. By utilizing luciferase reporters to monitor the T. atroviride core-clock, we confirmed the existence of circadian oscillations of ~26h that are temperature-compensated and modulated by environmental cues such as light and temperature. Notably, the presence of such rhythms appears to be highly dependent on the nutritional composition of the media. Heterologous expression of the T. atroviride negative clock component (tafrq) in a clock null (Δfrq) strain of Neurospora crassa restored core clock function in the latter fungus, with the same period observed in T. atroviride, confirming the role of tafrq as a bona fide core-clock component. Confrontation assays between wild-type and clock mutant strains of T. atroviride and B. cinerea, in constant light or darkness, revealed an inhibitory effect of light on T. atroviride's mycoparasitic capabilities. Interestingly, when confrontation assays were performed under light/dark cycles, T. atroviride's overgrowth capacity was enhanced when inoculations were at dawn compared to dusk. Deleting the core-clock negative element FRQ in B. cinerea, but not in T. atroviride, was vital for the daily differential phenotype, suggesting that the B. cinerea clock has a more significant influence on the result of this interaction. Additionally, we observed that T. atroviride clock components modulate development and secondary metabolism in this fungus, affecting the production of several molecules, including volatile compounds, such as 6-pentyl-α-pyrone (6-PP). Notably, we detected the rhythmic production of distinct T. atroviride volatile organic compounds (VOCs), which depended on its circadian clock. Thus, this study provides evidence on how clock components impact diverse aspects of T. atroviride lifestyle and how daily changes modulate fungal interactions and dynamics.

Data availability

All data generated and analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 1 and 2 and Table 1.

Article and author information

Author details

  1. Marlene Henríquez-Urrutia

    Molecular Genetics and Microbiology department, Pontificia Universidad Católica de Chile, Santiago, Chile
    Competing interests
    No competing interests declared.
  2. Rebecca Spanner

    Molecular Genetics and Microbiology department, Pontificia Universidad Católica de Chile, Santiago, Chile
    Competing interests
    No competing interests declared.
  3. Consuelo Olivares-Yánez

    Millennium Science Initiative Program, Millennium Institute for Integrative Biology, Santiago, Chile
    Competing interests
    No competing interests declared.
  4. Aldo Seguel-Avello

    Molecular Genetics and Microbiology department, Pontificia Universidad Católica de Chile, Santiago, Chile
    Competing interests
    No competing interests declared.
  5. Rodrigo Pérez-Lara

    Molecular Genetics and Microbiology department, Pontificia Universidad Católica de Chile, Santiago, Chile
    Competing interests
    No competing interests declared.
  6. Hector Guillén-Alonso

    Department of Biotechnology and Biochemistry, Cinvestav Unidad Irapuato, Irapuato, Mexico
    Competing interests
    No competing interests declared.
  7. Robert Winkler

    Department of Biotechnology and Biochemistry, Cinvestav Unidad Irapuato, Irapuato, Mexico
    Competing interests
    No competing interests declared.
  8. Alfredo Heriberto Herrera-Estrella

    Laboratorio de expresión génica y desarrollo en hongos, Unidad de Genómica Avanzada - Langebio, Irapuato, Mexico
    Competing interests
    No competing interests declared.
  9. Paulo Canessa

    Centro de Biotecnología Vegetal, Universidad Andrés Bello, Santiago, Chile
    For correspondence
    paulo.canessa@unab.cl
    Competing interests
    No competing interests declared.
  10. Luis F Larrondo

    Molecular Genetics and Microbiology department, Pontificia Universidad Católica de Chile, Santiago, Chile
    For correspondence
    llarrondo@bio.puc.cl
    Competing interests
    Luis F Larrondo, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8832-7109

Funding

Agencia Nacional de Investigación y Desarrollo (FONDECYT Regular 1211715)

  • Luis F Larrondo

Agencia Nacional de Investigación y Desarrollo (FONDECYT Postdoc 3180328)

  • Aldo Seguel-Avello

Agencia Nacional de Investigación y Desarrollo (FONDECYT Postdoc 3190628)

  • Consuelo Olivares-Yánez

Howard Hughes Medical Institute (the International Research Scholar program)

  • Luis F Larrondo

Agencia Nacional de Investigación y Desarrollo (Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio ICN17_022)

  • Paulo Canessa
  • Luis F Larrondo

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Antonis Rokas, Vanderbilt University, United States

Version history

  1. Received: June 17, 2021
  2. Accepted: August 10, 2022
  3. Accepted Manuscript published: August 11, 2022 (version 1)
  4. Version of Record published: August 30, 2022 (version 2)

Copyright

© 2022, Henríquez-Urrutia et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,255
    views
  • 338
    downloads
  • 8
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Marlene Henríquez-Urrutia
  2. Rebecca Spanner
  3. Consuelo Olivares-Yánez
  4. Aldo Seguel-Avello
  5. Rodrigo Pérez-Lara
  6. Hector Guillén-Alonso
  7. Robert Winkler
  8. Alfredo Heriberto Herrera-Estrella
  9. Paulo Canessa
  10. Luis F Larrondo
(2022)
Circadian oscillations in Trichoderma atroviride and the role of core clock components in secondary metabolism, development, and mycoparasitism against the phytopathogen Botrytis cinerea
eLife 11:e71358.
https://doi.org/10.7554/eLife.71358

Share this article

https://doi.org/10.7554/eLife.71358

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression
    Lucie Crhak Khaitova, Pavlina Mikulkova ... Karel Riha
    Research Article

    Heat stress is a major threat to global crop production, and understanding its impact on plant fertility is crucial for developing climate-resilient crops. Despite the known negative effects of heat stress on plant reproduction, the underlying molecular mechanisms remain poorly understood. Here, we investigated the impact of elevated temperature on centromere structure and chromosome segregation during meiosis in Arabidopsis thaliana. Consistent with previous studies, heat stress leads to a decline in fertility and micronuclei formation in pollen mother cells. Our results reveal that elevated temperature causes a decrease in the amount of centromeric histone and the kinetochore protein BMF1 at meiotic centromeres with increasing temperature. Furthermore, we show that heat stress increases the duration of meiotic divisions and prolongs the activity of the spindle assembly checkpoint during meiosis I, indicating an impaired efficiency of the kinetochore attachments to spindle microtubules. Our analysis of mutants with reduced levels of centromeric histone suggests that weakened centromeres sensitize plants to elevated temperature, resulting in meiotic defects and reduced fertility even at moderate temperatures. These results indicate that the structure and functionality of meiotic centromeres in Arabidopsis are highly sensitive to heat stress, and suggest that centromeres and kinetochores may represent a critical bottleneck in plant adaptation to increasing temperatures.

    1. Cell Biology
    Wan-ping Yang, Mei-qi Li ... Qian-qian Luo
    Research Article

    High-altitude polycythemia (HAPC) affects individuals living at high altitudes, characterized by increased red blood cells (RBCs) production in response to hypoxic conditions. The exact mechanisms behind HAPC are not fully understood. We utilized a mouse model exposed to hypobaric hypoxia (HH), replicating the environmental conditions experienced at 6000 m above sea level, coupled with in vitro analysis of primary splenic macrophages under 1% O2 to investigate these mechanisms. Our findings indicate that HH significantly boosts erythropoiesis, leading to erythrocytosis and splenic changes, including initial contraction to splenomegaly over 14 days. A notable decrease in red pulp macrophages (RPMs) in the spleen, essential for RBCs processing, was observed, correlating with increased iron release and signs of ferroptosis. Prolonged exposure to hypoxia further exacerbated these effects, mirrored in human peripheral blood mononuclear cells. Single-cell sequencing showed a marked reduction in macrophage populations, affecting the spleen’s ability to clear RBCs and contributing to splenomegaly. Our findings suggest splenic ferroptosis contributes to decreased RPMs, affecting erythrophagocytosis and potentially fostering continuous RBCs production in HAPC. These insights could guide the development of targeted therapies for HAPC, emphasizing the importance of splenic macrophages in disease pathology.