Effectors of Plant Necrotrophic Fungi Shao, Dandan; Smith, Damon L; Kabbage, Mehdi ...
Frontiers in plant science,
06/2021, Letnik:
12
Journal Article
Recenzirano
Odprti dostop
Plant diseases caused by necrotrophic fungal pathogens result in large economic losses in field crop production worldwide. Effectors are important players of plant-pathogen interaction and deployed ...by pathogens to facilitate plant colonization and nutrient acquisition. Compared to biotrophic and hemibiotrophic fungal pathogens, effector biology is poorly understood for necrotrophic fungal pathogens. Recent bioinformatics advances have accelerated the prediction and discovery of effectors from necrotrophic fungi, and their functional context is currently being clarified. In this review we examine effectors utilized by necrotrophic fungi and hemibiotrophic fungi in the latter stages of disease development, including plant cell death manipulation. We define "effectors" as secreted proteins and other molecules that affect plant physiology in ways that contribute to disease establishment and progression. Studying and understanding the mechanisms of necrotrophic effectors is critical for identifying avenues of genetic intervention that could lead to improved resistance to these pathogens in plants.
Soybean vein necrosis virus (SVNV; genus Tospovirus; Family Bunyaviridae) is a negative-sense single-stranded RNA virus that has been detected across the United States and in Ontario, Canada. In ...2013, a seed lot of a commercial soybean variety (Glycine max) with a high percentage of discolored, deformed and undersized seed was obtained. A random sample of this seed was planted in a growth room under standard conditions. Germination was greater than 90% and the resulting seedlings looked normal. Four composite samples of six plants each were tested by reverse transcription polymerase chain reaction (RT-PCR) using published primers complimentary to the S genomic segment of SVNV. Two composite leaflet samples retrieved from seedlings yielded amplicons with a size and sequence predictive of SVNV. Additional testing of twelve arbitrarily selected individual plants resulted in the identification of two SVNV positive plants. Experiments were repeated by growing seedlings from the same seed lot in an isolated room inside a thrips-proof cage to further eliminate any external source of infection. Also, increased care was taken to reduce any possible PCR contamination. Three positive plants out of forty-eight were found using these measures. Published and newly designed primers for the L and M RNAs of SVNV were also used to test the extracted RNA and strengthen the diagnosis of viral infection. In experiments, by three scientists, in two different labs all three genomic RNAs of SVNV were amplified in these plant materials. RNA-seq analysis was also conducted using RNA extracted from a composite seedling sample found to be SVNV-positive and a symptomatic sample collected from the field. This analysis revealed both sense and anti-sense reads from all three gene segments in both samples. We have shown that SVNV can be transmitted in seed to seedlings from an infected seed lot at a rate of 6%. To our knowledge this is the first report of seed-transmission of a Tospovirus.
While fungal biotrophs are dependent on successfully suppressing/subverting host defenses during their interaction with live cells, necrotrophs, due to their lifestyle are often confronted with a ...suite of toxic metabolites. These include an assortment of plant defense compounds (PDCs) which can demonstrate broad antifungal activity. These PDCs can be either constitutively present in plant tissue or induced in response to infection, but are nevertheless an important obstacle which needs to be overcome for successful pathogenesis. Fungal necrotrophs have developed a number of strategies to achieve this goal, from the direct detoxification of these compounds through enzymatic catalysis and modification, to the active transport of various PDCs to achieve toxin sequestration and efflux. Studies have shown across multiple pathogens that the efficient detoxification of host PDCs is both critical for successful infection and often a determinant factor in pathogen host range. Here, we provide a broad and comparative overview of the various mechanisms for PDC detoxification which have been identified in both fungal necrotrophs and fungal pathogens which depend on detoxification during a necrotrophic phase of infection. Furthermore, the effect that these mechanisms have on fungal host range, metabolism, and disease control will be discussed.
Sclerotinia sclerotiorum is a broad-host range necrotrophic pathogen which is the causative agent of Sclerotinia stem rot (SSR), and a major disease of soybean (Glycine max). A time course ...transcriptomic analysis was performed in both compatible and incompatible soybean lines to identify pathogenicity and developmental factors utilized by S. sclerotiorum to achieve pathogenic success.
A comparison of genes expressed during early infection identified the potential importance of toxin efflux and nitrogen metabolism during the early stages of disease establishment. The later stages of infection were characterized by an apparent shift to survival structure formation. Analysis of genes highly upregulated in-planta revealed a temporal regulation of hydrolytic and detoxification enzymes, putative secreted effectors, and secondary metabolite synthesis genes. Redox regulation also appears to play a key role during the course of infection, as suggested by the high expression of genes involved in reactive oxygen species production and scavenging. Finally, distinct differences in early gene expression were noted based on the comparison of S. sclerotiorum infection of resistant and susceptible soybean lines.
Although many potential virulence factors have been noted in the S. sclerotiorum pathosystem, this study serves to highlight soybean specific processes most likely to be critical in successful infection. Functional studies of genes identified in this work are needed to confirm their importance to disease development, and may constitute valuable targets of RNAi approaches to improve resistance to SSR.
Corn silage, made from
, is a high-energy feed that is important for feeding dairy cows. Plant diseases, such as those caused by
, can decrease silage corn yields and quality.
(teleomorph
) is an ...ascomycete fungus that causes Gibberella ear and stalk rot in corn.
produces deoxynivalenol (DON), a secondary metabolite toxic to humans and animals. An understanding of the distribution of DON and
throughout the corn plant is important for determining the quality of corn silage. A partitioned sample experiment that included two brown midrib silage hybrids and three fungicide treatments was conducted in research plots located in Arlington, WI, U.S.A., in 2018 and 2019. At harvest, stalk and ear parts were physically separated, dried, and ground for analysis. DON concentration (in parts per million) was determined using an enzyme-linked immunosorbent assay, and
DNA concentration (in picograms per nanogram) was determined using quantitative PCR. DON and
DNA were detected in all samples, demonstrating accumulation of the fungus in stalks and ears of the plant. In 2018, DON contamination was as high as 30 ppm and varied drastically between stalks and ears. In 2019, DON concentrations were much lower (<5 ppm), but were consistently higher in stalk samples than ear samples. Across all samples, DON concentrations and
accumulation were highly correlated within the separated stalk (
= 0.78) and ear portions (
= 0.87) but were not correlated between ears and stalks. Depending on the weather and planting conditions in a given year, stalk infections or ear infections may occur by
, leading to subsequent DON increases in those respective parts that are independent of each other.
commonly causes Fusarium head blight on wheat, barley, rice, and oats.
produces nivalenol (NIV) and deoxynivalenol (DON) and forms derivatives of DON based on its acetylation sites. The fungus is ...profiled into chemotypes based on DON derivative chemotypes (3 acetyldeoxynivalenol 3ADON chemotype; 15 acetyldeoxynivalenol 15ADON chemotype), and/or the NIV chemotype. This study assessed the
population found on wheat and the chemotype profile of the isolates collected from 2016 and 2017 in Wisconsin.
was isolated from all locations sampled in both 2016 and 2017.
was isolated only from Door County in 2016. Over both growing seasons, 91% of isolates were identified as the 15ADON chemotype, while 9% of isolates were identified as the 3ADON chemotype. Aggressiveness was quantified by area under the disease progress curve (AUDPC). The isolates with the highest AUDPC values were from the highest wheat-producing cropping districts in the state. Deoxynivalenol production in grain and sporulation and growth rate in vitro were compared with aggressiveness in the greenhouse. Our results showed that 3ADON isolates in Wisconsin were among the highest in sporulation capacity, growth rate, and DON production in grain. However, there were no significant differences in aggressiveness between the 3ADON and 15ADON isolates. The results of this research detail the baseline frequency and distribution of 3ADON and 15ADON chemotypes observed in Wisconsin. Chemotype distributions within populations of
in Wisconsin should continue to be monitored in the future.
Summary
Sclerotinia sclerotiorum, a predominately necrotrophic fungal pathogen with a broad host range, causes a significant yield‐limiting disease of soybean called Sclerotinia stem rot. Resistance ...mechanisms against this pathogen in soybean are poorly understood, thus hindering the commercial deployment of resistant varieties. We used a multiomic approach utilizing RNA‐sequencing, gas chromatography–mass spectrometry‐based metabolomics and chemical genomics in yeast to decipher the molecular mechanisms governing resistance to S. sclerotiorum in soybean. Transcripts and metabolites of two soybean recombinant inbred lines, one resistant and one susceptible to S. sclerotiorum were analysed in a time course experiment. The combined results show that resistance to S. sclerotiorum in soybean is associated in part with an early accumulation of JA‐Ile ((+)‐7‐iso‐jasmonoyl‐L‐isoleucine), a bioactive jasmonate, increased ability to scavenge reactive oxygen species, and importantly, a reprogramming of the phenylpropanoid pathway leading to increased antifungal activities. Indeed, we noted that phenylpropanoid pathway intermediates, such as 4‐hydroxybenzoate, cinnamic acid, ferulic acid and caffeic acid, were highly accumulated in the resistant line. In vitro assays show that these metabolites and total stem extracts from the resistant line clearly affect S. sclerotiorum growth and development. Using chemical genomics in yeast, we further show that this antifungal activity targets ergosterol biosynthesis in the fungus, by disrupting enzymes involved in lipid and sterol biosynthesis. Overall, our results are consistent with a model where resistance to S. sclerotiorum in soybean coincides with an early recognition of the pathogen, leading to the modulation of the redox capacity of the host and the production of antifungal metabolites.
Sclerotinia stem rot (SSR), caused by
Sclerotinia sclerotiorum
, is one of the most important diseases of soybean. Disease management is complicated by the long-term survival of sclerotia in the soil ...and the absence of resistance in elite, commercial cultivars. Furthermore, the lifecycle of
S. sclerotiorum
in soybean fields is highly dependent on weather conditions, leading to a highly sporadic occurrence of the disease over seasons and an aggregated distribution within fields. Management relies on a multi-pronged approach of combining partially resistant cultivars with cultural practices, such as altering row spacing and planting population, along with chemical control. These control measures are constrained by economic trade-offs, incomplete efficacy of chemicals, and a lack of understanding of application timing for fungicides. Newer tools have been developed to improve management, such as disease prediction models that can assist farmers in making decisions about fungicide application. This review aims to introduce the
Sclerotinia
pathosystem in soybean, while covering the complicated biology of
S. sclerotiorum
that leads to the need for integrated management by soybean farmers.
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Development of new additive manufacturing materials often requires the production of several batches of relatively large volumes in order to print and test objects. This can be ...difficult for many materials that are expensive or difficult to produce in large volumes on the laboratory scale. Bioprinter systems are advantageous in this regard, however, commercial systems are expensive or do not have the ability to use photopolymers. Herein, we outline a Syringe Pump Extruder and Curing System (SPECS) modification for inexpensive filament-based 3D printers which enables the use of standard bioplotter materials and photopolymers. The system is capable of using multiple syringe volumes and needle sizes that can be quickly and easily exchanged. The SPECS modification is demonstrated using a Prusa i3 mk3 fused filament fabrication printer to print several 3D objects and films using stereolithography (SLA) photopolymer resin. Geometric accuracy in the X, Y, and Z directions was ±0.1 mm using a 5 ml syringe, 22-gauge needle, and commercial SLA resin. The SPECS system could be of great benefit for laboratories pursing material development in the area of additive manufacturing.
Genome-wide association (GWAS) and epistatic (GWES) studies along with expression studies in soybean Glycine max (L.) Merr. were leveraged to dissect the genetics of Sclerotinia stem rot (SSR) caused ...by Sclerotinia sclerotiorum (Lib.) de Bary, a significant fungal disease causing yield and quality losses. A large association panel of 466 diverse plant introduction accessions were phenotyped in multiple field and controlled environments to: (1) discover sources of resistance, (2) identify SNPs associated with resistance, and (3) determine putative candidate genes to elucidate the mode of resistance. We report 58 significant main effect loci and 24 significant epistatic interactions associated with SSR resistance, with candidate genes involved in a wide range of processes including cell wall structure, hormone signaling, and sugar allocation related to plant immunity, revealing the complex nature of SSR resistance. Putative candidate genes for example, PHYTOALEXIN DEFFICIENT 4 (PAD4), ETHYLENE-INSENSITIVE 3-LIKE 1 (EIL3), and ETHYLENE RESPONSE FACTOR 1 (ERF1) clustered into salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) pathways suggest the involvement of a complex hormonal network typically activated by both necrotrophic (ET/JA) and biotrophic (SA) pathogens supporting that S. sclerotiorum is a hemibiotrophic plant pathogen.
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