Papers by Ulrich Schaffrath
Plant Physiology, 2014
For plant pathogenic fungi, such as powdery mildews, that survive only on a limited number of hos... more For plant pathogenic fungi, such as powdery mildews, that survive only on a limited number of host plant species, it is a matter of vital importance that their spores sense that they landed on the right spot to initiate germination as quickly as possible. We investigated a barley (Hordeum vulgare) mutant with reduced epicuticular leaf waxes on which spores of adapted and nonadapted powdery mildew fungi showed reduced germination. The barley gene responsible for the mutant wax phenotype was cloned in a forward genetic screen and identified to encode a 3-KETOACYL-CoA SYNTHASE (HvKCS6), a protein participating in fatty acid elongation and required for synthesis of epicuticular waxes. Gas chromatography-mass spectrometry analysis revealed that the mutant has significantly fewer aliphatic wax constituents with a chain length above C-24. Complementation of the mutant restored wild-type wax and overcame germination penalty, indicating that wax constituents less present on the mutant are a crucial clue for spore germination. Investigation of Arabidopsis (Arabidopsis thaliana) transgenic plants with sense silencing of Arabidopsis REQUIRED FOR CUTICULAR WAX PRODUCTION1, the HvKCS6 ortholog, revealed the same germination phenotype against adapted and nonadapted powdery mildew fungi. Our findings hint to an evolutionary conserved mechanism for sensing of plant surfaces among distantly related powdery mildews that is based on KCS6-derived wax components. Perception of such a signal must have been evolved before the monocot-dicot split took place approximately 150 million years ago.
Molecular Plant-Microbe Interactions, 2006
the induction of a unique physiological state called "priming." The primed state can also be indu... more the induction of a unique physiological state called "priming." The primed state can also be induced by treatment of plants with various natural and synthetic compounds. Primed plants display either faster, stronger, or both activation of the various cellular defense responses that are induced following attack by either pathogens or insects or in response to abiotic stress. Although the phenomenon has been known for decades, most progress in our understanding of priming has been made over the past few years. Here, we summarize the current knowledge of priming in various induced-resistance phenomena in plants.
Plant Cell Reports, 2016
Adapted pathogens are able to modulate cell responses of their hosts most likely due to the activ... more Adapted pathogens are able to modulate cell responses of their hosts most likely due to the activity of secreted effector molecules thereby enabling colonisation by ostensible nonhost pathogens. It is postulated that host and nonhost pathogens of a given plant species differ in their repertoire of secreted effector molecules that are able to suppress plant resistance. We pursued the strategy of identifying novel effectors of Magnaporthe oryzae, the causal agent of blast disease, by comparing the infection process of closely related host vs. nonhost Magnaporthe species on barley (Hordeum vulgare L.). When both types of pathogen simultaneously attacked the same cell, the nonhost isolate became a successful pathogen possibly due to potent effectors secreted by the host isolate. Microarray studies led to a set of M. oryzae Hypothetical Effector Genes (MoHEGs) which were classified as Early- and LateMoHEGs according to the maximal transcript abundance during colonization of barley. Interestingly, orthologs of these MoHEGs from a nonhost pathogen were similarly regulated when investigated in a host situation, suggesting evolutionary conserved functions. Knockout mutants of MoHEG16 from the group of EarlyMoHEGs were less virulent on barley and microscopic studies revealed an attenuated transition from epidermal to mesophyll colonization. MoHEG13, a LateMoHEG, was shown to antagonize cell death induced by M. oryzae Necrosis-and ethylene-inducing-protein-1 (Nep1)-like proteins in Nicotiana benthamiana. MoHEG13 has a virulence function as a knockout mutant showed attenuated disease progression when inoculated on barley.
Physiological and Molecular Plant Pathology, 2004
The acquired resistance state of rice plants, rendering them resistant to subsequent infections w... more The acquired resistance state of rice plants, rendering them resistant to subsequent infections with virulent pathogens, can be triggered by treatments with non-host pathogens, necrotizing pathogens and certain chemicals. The 13-lipoxygenase RCI1 (LOX2:Os:2) is induced specifically following treatment with chemical inducers of resistance. Here, we report that the over-expression of RCI1 in a transient transformation assay leads to PR1 transcript accumulation in rice leaves. In addition, we show that this property is due to the enzymatic activity of the transgenic protein. Furthermore, exogenous application of (13S,9Z,11E,15Z)-13-hydroxy-9,11,15-octadecatrienoic acid (13-HOT), an oxylipin deriving from the reductase branch of the lipoxygenase pathway, is capable of inducing PR1. These results support a role for RCI1 in the generation of signalling molecules during the establishment of the acquired resistance response in rice. q
Frontiers in plant science, 2015
Non-host resistance (NHR) is the resistance of plants to a plethora of non-adapted pathogens and ... more Non-host resistance (NHR) is the resistance of plants to a plethora of non-adapted pathogens and is considered as one of the most robust resistance mechanisms of plants. Studies have shown that the efficiency of resistance in general and NHR in particular could vary in different plant organs, thus pointing to tissue-specific determinants. This was exemplified by research on host and non-host interactions of the fungal plant pathogen Magnaporthe oryzae with rice and Arabidopsis, respectively. Thus, rice roots were shown to be impaired in resistance to M. oryzae isolates to which leaves of the same rice cultivar are highly resistant. Moreover, roots of Arabidopsis are also accessible to penetration by M. oryzae while leaves of this non-host plant cannot be infected. We addressed the question whether or not other plant tissues such as the reproductive system also differ in NHR compared to leaves. Inoculation experiments on wheat with different Magnaporthe species forming either a host ...
Molecular Plant Microbe Interactions Mpmi, Sep 1, 2006
Magnaporthe oryzae is a major pathogen of rice (Oryza sativa L.) but is also able to infect other... more Magnaporthe oryzae is a major pathogen of rice (Oryza sativa L.) but is also able to infect other grasses, including barley (Hordeum vulgare L.). Here, we report a study using Magnaporthe isolates collected from other host plant species to evaluate their capacity to infect barley. A nonhost type of resistance was detected in barley against isolates derived from genera Pennisetum (fontaingrass) or Digitaria (crabgrass), but no resistance occurred in response to isolates from rice, genus Eleusine (goosegrass), wheat (Triticum aestivum L.), or maize (Zea mays L.), respectively. Restriction of pathogen growth in the nonhost interaction was investigated microscopically and compared with compatible interactions. Real-time polymerase chain reaction was used to quantify fungal biomass in both types of interaction. The phylogenetic relationship among the Magnaporthe isolates used in this study was investigated by inferring gene trees for fragments of three genes, actin, calmodulin, and βtubulin. Based on phylogenetic analysis, we could distinguish different species that were strictly correlated with the ability of the isolates to infect barley. We demonstrated that investigating specific host interaction phenotypes for a range of pathogen isolates can accurately highlight genetic diversity within a pathogen population.
Molecular Plant, 2015
Modular proteins are an evolutionary answer to optimize performance of proteins that physically i... more Modular proteins are an evolutionary answer to optimize performance of proteins that physically interact with each other for functionality. Using a combination of genetic and biochemical experiments, we characterized the rice protein OsJAC1 which consists of a jacalin-related lectin (JRL) domain predicted to bind mannose-containing oligosaccharides and a dirigent domain which might function in stereoselective coupling of monolignols. Transgenic overexpression of OsJAC1 in rice resulted in quantitative broad-spectrum resistance against different pathogens including bacteria, oomycetes and fungi. Overexpression of this gene or its wheat ortholog TAJA1 in barley enhanced resistance against the powdery mildew fungus. Both protein domains of OsJAC1 are required to establish resistance as indicated by single or combined transient expression of individual domains. Expression of artificially separated and fluorescence-tagged protein domains showed that the JRL domain is sufficient for targeting the powdery mildew penetration site. Nevertheless, co-localization of lectin- and dirigent-domain occurred. Phylogenetic analyses revealed orthologs of OsJAC1 exclusively within the Poaceae plant family. Dicots, by contrast, only contain proteins with either JRL or dirigent domain(s). Altogether, our results identify OsJAC1 as a representative of a novel type of resistance proteins derived from a plant-lineage specific gene fusion event for better function in local pathogen defense.
Physiological and Molecular Plant Pathology, 1995
Plant biotechnology journal, Jan 15, 2015
The wheat gene Lr34 confers durable and partial field resistance against the obligate biotrophic,... more The wheat gene Lr34 confers durable and partial field resistance against the obligate biotrophic, pathogenic rust fungi and powdery mildew in adult wheat plants. The resistant Lr34 allele evolved after wheat domestication through two gain-of-function mutations in an ATP-binding cassette transporter gene. An Lr34-like fungal disease resistance with a similar broad-spectrum specificity and durability has not been described in other cereals. Here, we transformed the resistant Lr34 allele into the japonica rice cultivar Nipponbare. Transgenic rice plants expressing Lr34 showed increased resistance against multiple isolates of the hemibiotrophic pathogen Magnaporthe oryzae, the causal agent of rice blast disease. Host cell invasion during the biotrophic growth phase of rice blast was delayed in Lr34-expressing rice plants, resulting in smaller necrotic lesions on leaves. Lines with Lr34 also developed a typical, senescence-based leaf tip necrosis (LTN) phenotype. Development of LTN durin...
The New phytologist, 2014
Asian soybean rust (Phakopsora pachyrhizi) causes a devastating disease in soybean (Glycine max).... more Asian soybean rust (Phakopsora pachyrhizi) causes a devastating disease in soybean (Glycine max). We tested the hypothesis that the fungus generates high turgor pressure in its hyaline appressoria to mechanically pierce epidermal cells. Turgor pressure was determined by a microscopic technique, called transmitted light double-beam interference Mach-Zehnder microscopy (MZM), which was developed in the 1960s as a forefront of live cell imaging. We revitalized some original microscopes and equipped them for modern image capturing. MZM data were corroborated by cytorrhysis experiments. Incipient cytorrhysis determined the turgor pressure in appressoria of P. pachyrhizi to be equivalent to 5.13 MPa. MZM data revealed that osmotically active sugar alcohols only accounted for 75% of this value. Despite having a lower turgor pressure, hyaline rust appressoria were able to penetrate non-biodegradable polytetrafluoroethylene (PTFE) membranes more efficiently than do melanized appressoria of t...
BMC Plant Biology, 2015
Background: Plant hormones are well known regulators which balance plant responses to abiotic and... more Background: Plant hormones are well known regulators which balance plant responses to abiotic and biotic stresses. We investigated the role of abscisic acid (ABA) in resistance of barley (Hordeum vulgare L.) against the plant pathogenic fungus Magnaporthe oryzae. Results: Exogenous application of ABA prior to inoculation with M. oryzae led to more disease symptoms on barley leaves. This result contrasted the finding that ABA application enhances resistance of barley against the powdery mildew fungus. Microscopic analysis identified diminished penetration resistance as cause for enhanced susceptibility. Consistently, the barley mutant Az34, impaired in ABA biosynthesis, was less susceptible to infection by M. oryzae and displayed elevated penetration resistance as compared to the isogenic wild type cultivar Steptoe. Chemical complementation of Az34 mutant plants by exogenous application of ABA re-established disease severity to the wild type level. The role of ABA in susceptibility of barley against M. oryzae was corroborated by showing that ABA application led to increased disease severity in all barley cultivars under investigation except for the most susceptible cultivar Pallas. Interestingly, endogenous ABA concentrations did not significantly change after infection of barley with M. oryzae. Conclusion: Our results revealed that elevated ABA levels led to a higher disease severity on barley leaves to M. oryzae. This supports earlier reports on the role of ABA in enhancing susceptibility of rice to the same pathogen and thereby demonstrates a host plant-independent function of this phytohormone in pathogenicity of monocotyledonous plants against M. oryzae.
Plant Signaling & Behavior, 2011
The acquired resistance state of rice plants, rendering them resistant to subsequent infections w... more The acquired resistance state of rice plants, rendering them resistant to subsequent infections with virulent pathogens, can be triggered by treatments with non-host pathogens, necrotizing pathogens and certain chemicals. The 13-lipoxygenase RCI1 (LOX2:Os:2) is induced specifically following treatment with chemical inducers of resistance. Here, we report that the over-expression of RCI1 in a transient transformation assay leads to PR1 transcript accumulation in rice leaves. In addition, we show that this property is due to the enzymatic activity of the transgenic protein. Furthermore, exogenous application of (13S,9Z,11E,15Z)-13-hydroxy-9,11,15-octadecatrienoic acid (13-HOT), an oxylipin deriving from the reductase branch of the lipoxygenase pathway, is capable of inducing PR1. These results support a role for RCI1 in the generation of signalling molecules during the establishment of the acquired resistance response in rice. q
The Rir1b gene of rice (Oryza sativa) is one of a set of putative defense genes whose transcripts... more The Rir1b gene of rice (Oryza sativa) is one of a set of putative defense genes whose transcripts accumulate upon inoculation of rice with the non-host pathogen Pseudomonas syringae pv. syringae. It belongs to a family of genes encoding small extracellular proteins so far only identified in cereals. To assess the function of the Rir1b gene in rice blast resistance, it was placed under the control of the CaMV 35S promoter and transferred into rice plants of the japonica variety Taipei 309 by biolistic transformation of immature embryos. Two out of 12 hygromycin-resistant regenerated plants (OE1 and OE3) were fertile. DNA gel blot analysis suggested that these two T 0 plants were independent transformants, each of which had stably incorporated one complete copy of the transgene into the genome. In addition, the OE1 plant appeared also a contain a rearranged copy or incomplete copy. T 1 plants homozygous for the transgene were identified by DNA gel blot analysis of individual T 2 progeny and further propagated. Expression analysis of the transgene showed that the transgene was active both in T 1 plants and homozygous decendants. Challenge inoculation of homozygous transgenic plants with Magnaporthe grisea, the causal agent of rice blast disease, revealed that both independent transgenic lines were more resistant than the untransformed wild type, suggesting that over-expression of the Rir1b gene confers partial resistance against this important pathogen.
Soybean - Biochemistry, Chemistry and Physiology, 2011
The basidiomycete Phakopsora pachyrhizi (P. pachyrhizi) causes Asian soybean rust, one of the mos... more The basidiomycete Phakopsora pachyrhizi (P. pachyrhizi) causes Asian soybean rust, one of the most devastating plant diseases on soybean. When inoculated on the nonhost barley P. pachyrhizi caused only very small necrotic spots, typical for an incompatible interaction, which involves a hypersensitive cell death reaction. A microscopic inspection of the interaction of barley with P. pachyrhizi revealed that the fungus germinated on barley and formed functional appressoria on epidermal cells. The fungus attempted to directly penetrate through periclinal cell walls but often failed, arrested in plant cell wall appositions that stained positively for callose. Penetration resistance depends on intact ROR1(REQUIRED FOR mlo-SPECIFIED RESISTANCE 1) and ROR2 genes of barley. If the fungus succeeded in penetration, epidermal cell death took place. Dead epidermal cells did not generally restrict fungal development but allowed for mesophyll invasion, which was followed by mesophyll cell death and fungal arrest. Transient or stable over expression of the barley cell death suppressor BAX inhibitor-1 reduced both epidermal cell death and fungal penetration success. Data suggest that P. pachyrhizi provokes a programmed cell death facilitating fungal entry into epidermal cells of barley.
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Papers by Ulrich Schaffrath