skip to main content


Title: SUMOylation regulates low‐temperature survival and oxidative DNA damage tolerance in Botrytis cinerea
Summary

SUMOylation as one of the protein post‐translational modifications plays crucial roles in multiple biological processes of eukaryotic organisms.Botrytis cinereais a devastating fungal pathogen and capable of infecting plant hosts at low temperature. However, the molecular mechanisms of low‐temperature adaptation are largely unknown in fungi.

Combining with biochemical methods and biological analyses, we report that SUMOylation regulates pathogen survival at low temperature and oxidative DNA damage response during infection inB. cinerea. The heat shock protein (Hsp70) BcSsb and E3 ubiquitin ligase BcRad18 were identified as substrates of SUMOylation; moreover, their SUMOylation both requires a single unique SUMO‐interacting motif (SIM).

SUMOylated BcSsb regulates β‐tubulin accumulation, thereby affecting the stability of microtubules and consequently mycelial growth at low temperature. On the contrary, SUMOylated BcRad18 modulates mono‐ubiquitination of the sliding clamp protein proliferating cell nuclear antigen (PCNA), which is involved in response to oxidative DNA damage during infection.

Our study uncovers the molecular mechanisms of SUMOylation‐mediated low‐temperature survival and oxidative DNA damage tolerance during infection in a devastating fungal pathogen, which provides novel insights into low‐temperature adaptation and pathogenesis for postharvest pathogens as well as new targets for inhibitor invention in disease control.

 
more » « less
NSF-PAR ID:
10395836
Author(s) / Creator(s):
 ;  ;  ;  ;  ;  ;  ;  
Publisher / Repository:
Wiley-Blackwell
Date Published:
Journal Name:
New Phytologist
Volume:
238
Issue:
2
ISSN:
0028-646X
Page Range / eLocation ID:
p. 817-834
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Summary

    Fungal phytopathogens can suppress plant immune mechanisms in order to colonize living host cells. Identifying all the molecular components involved is critical for elaborating a detailed systems‐level model of plant infection probing pathogen weaknesses; yet, the hierarchy of molecular events controlling fungal responses to the plant cell is not clear.

    Here we show how, in the blast fungusMagnaporthe oryzae, terminating rice innate immunity requires a dynamic network of redox‐responsive E3 ubiquitin ligases targeting fungal sirtuin 2 (Sir2), an antioxidation regulator required for suppressing the host oxidative burst.

    Immunoblotting, immunopurification, mass spectrometry and gene functional analyses showed that Sir2 levels responded to oxidative stress via a mechanism involving ubiquitination and three antagonistic E3 ubiquitin ligases: Grr1 and Ptr1 maintained basal Sir2 levels in the absence of oxidative stress; Upl3 facilitated Sir2 accumulation in response to oxidative stress. Grr1 and Upl3 interacted directly with Sir2 in a manner that decreased and scaled with oxidative stress, respectively.

    DeletingUPL3depleted Sir2 during growth in rice cells, triggering host immunity and preventing infection. OverexpressingSIR2in the Δupl3mutant remediated pathogenicity. Our work reveals how redox‐responsive E3 ubiquitin ligases inM. oryzaemediate Sir2 accumulation‐dependent antioxidation to modulate plant innate immunity and host susceptibility.

     
    more » « less
  2. Summary

    Histone lysine methylations (HLMs) are implicated in control of gene expression in different eukaryotes. However, the role of HLMs in regulating desirable crop traits and the enzymes involved in these modifications are poorly understood.

    We studied the functions of tomato histone H3 lysine methyltransferases SET Domain Group 33 (SDG33) and SDG34 in biotic and abiotic stress responses.SDG33andSDG34gene edited mutants were altered in H3K36 and H3K4 methylations, and expression of genes involved in diverse processes and responses to biotic and abiotic stimuli.

    The double but not the single mutants show resistance to the fungal pathogenBotrytis cinerea.Interestingly, single mutants were tolerant to drought and the double mutant showed superior tolerance and plant growth consistent with independent and additive functions. Mutants maintained higher water status during drought and improved recovery and survival after lapse of drought.

    Notably, diminution of H3K4 and H3K36 trimethylation and expression of negative regulators in challenged plants contributes to stress tolerance of the mutants. Mutations inSDG33andSDG34are likely to remove predisposition to biotic and abiotic stress by disrupting permissive transcriptional context promoting expression of negative regulatory factors. These allows improvement of stress and pathogen tolerance, without growth trade‐offs, through modification of histone epigenetic marks.

     
    more » « less
  3. Abstract

    Resistance and tolerance are unique host defence strategies that can limit the impacts of a pathogen on a host. However, for most wildlife–pathogen systems, there are still fundamental uncertainties regarding (a) how changes in resistance and tolerance can affect disease outcomes and (b) the mechanisms underlying resistance and tolerance in host populations.

    Here, we first compared observed patterns of resistance and tolerance and their effects on disease outcomes among salamander species that are susceptible to infection and mortality from the emerging fungal pathogenBatrachochytrium salamandrivorans(Bsal). We then tested whether two putative mechanisms that contribute to host resistance and tolerance, skin sloughing and skin lesion reduction, predicted reducedBsalgrowth rate or increased host survival during infection, respectively.

    We performed multi‐doseBsalchallenge experiments on four species of Salamandridae found throughout North America. We combined the laboratory experiments with dynamic models and sensitivity analysis to examine how changes in load‐dependent resistance and tolerance functions affectedBsal‐induced mortality risk. Finally, we used our disease model to test whether skin sloughing and lesion reduction predicted variability in infection outcomes not described byBsalinfection intensity.

    We found that resistance and tolerance differed significantly among salamander species, with the most susceptible species being both less resistance and less tolerant ofBsalinfection. Our dynamic model showed that the relative influence of resistance versus tolerance on host survival was species‐dependent—increasing resistance was only more influential than increasing tolerance for the least tolerant species where changes in pathogen load had a threshold‐like effect on host survival. Testing two candidate mechanisms of resistance and tolerance, skin sloughing and lesion reduction, respectively, we found limited support that either of these processes were strong mechanisms of host defence.

    Our study contributes to a broader understanding of resistance and tolerance in host–pathogen systems by showing that differences in host tolerance can significantly affect whether changes in resistance or tolerance have larger effects on disease outcomes, highlighting the need for species and even population‐specific management approaches that target host defence strategies.

    A freePlain Language Summarycan be found within the Supporting Information of this article.

     
    more » « less
  4. Abstract

    Ophiocordycepsfungi manipulate the behaviour of their ant hosts to produce a summit disease phenotype, thereby establishing infected ant cadavers onto vegetation at elevated positions suitable for fungal growth and transmission. Multiple environmental and ecological factors have been proposed to shape the timing, positioning and outcome of these manipulations.

    We conducted a long‐term field study ofOphiocordyceps camponoti‐floridaniinfections ofCamponotus floridanusants—the Florida zombie ants. We propose and refine hypotheses on the factors that shape infection outcomes by tracking the occurrence of and fungal growth from hundreds of ant cadavers. We modelled and report these data in relation to weather, light, vegetation and attack by hyperparasites.

    We investigated environmental factors that could affect the occurrence and location of newly manipulated ant cadavers. New cadaver occurrence was preferentially biased towards epiphyticTillandsiabromeliads, canopy openness and summer weather conditions (an interactive effect of temperature, humidity and precipitation). Furthermore, we suggest that incident light at the individual cadaver level reflects microhabitat choice by manipulated ants or selective pressure on cadaver maintenance for conditions that improve fungal survival.

    We also asked which environmental conditions affect fungal fitness. Continued fungal development of reproductive structures and putative transmission increased with moist weather conditions (interaction of humidity and precipitation) and canopy openness, while being reduced by hyperparasitic mycoparasite infections. Moreover, under the most open canopy conditions, we found an atypicalOphiocordycepsgrowth morphology that could represent a plastic response to conditions influenced by high light levels.

    Taken together, we explore general trends and the effects of various ecological conditions on host and parasite disease outcomes in the Florida zombie ant system. These insights from the field can be used to inform experimental laboratory setups that directly test the effects of biotic and abiotic factors on fungus–ant interactions or aim to uncover underlying molecular mechanisms.

    Read the freePlain Language Summaryfor this article on the Journal blog.

     
    more » « less
  5. Summary

    Plant diseases are an important threat to food production. While major pathogenicity determinants required for disease have been extensively studied, less is known on how pathogens thrive during host colonization, especially at early infection stages.

    Here, we used randomly barcoded‐transposon insertion site sequencing (RB‐TnSeq) to perform a genome‐wide screen and identify key bacterial fitness determinants of the vascular pathogenXanthomonas campestrispvcampestris(Xcc) during infection of the cauliflower host plant (Brassica oleracea). This high‐throughput analysis was conducted in hydathodes, the natural entry site ofXcc, in xylem sap and in synthetic media.

    Xccdid not face a strong bottleneck during hydathode infection. In total, 181 genes important for fitness were identified in plant‐associated environments with functional enrichment in genes involved in metabolism but only few genes previously known to be involved in virulence. The biological relevance of 12 genes was independently confirmed by phenotyping single mutants. Notably, we show that XC_3388, a protein with no known function (DUF1631), plays a key role in the adaptation and virulence ofXccpossibly through c‐di‐GMP‐mediated regulation.

    This study revealed yet unsuspected social behaviors adopted byXccindividuals when confined inside hydathodes at early infection stages.

     
    more » « less