Recent progress in discovery and functional analysis of effector proteins of fungal and oomycete plant pathogens

Ellis, Jeffrey G.; Rafiqi, Maryam; Gan, Pamela; Chakrabarti, Apratim; Dodds, Peter N.

doi:10.1016/j.pbi.2009.05.004

Cited by 152 publications

(118 citation statements)

References 41 publications

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“…Effectoromics, a high-throughput functional genomics approach has recently emerged as a powerful tool to identify resistance (R) genes in crop plants and matching avirulence (Avr) genes of pathogens [1][2][3][4] . In contrast to the more time-consuming stable transformation with R genes, the effectoromics strategy is based on transient assays of pathogen gene sequences.…”

Section: Introductionmentioning

confidence: 99%

Agroinfiltration and PVX Agroinfection in Potato and <em>Nicotiana benthamiana</em>

Rietman

Vleeshouwers

2014

JoVE

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Agroinfiltration and PVX agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. The most commonly used agent for agroinfiltration is Agrobacterium tumefaciens, a pathogen of many dicot plant species. This implies that agroinfiltration can be applied to many plant species. Here, we present our protocols and expected results when applying these methods to the potato (Solanum tuberosum), its related wild tuber-bearing Solanum species (Solanum section Petota) and the model plant Nicotiana benthamiana. In addition to functional analysis of single genes, such as resistance (R) or avirulence (Avr) genes, the agroinfiltration assay is very suitable for recapitulating the R-AVR interactions associated with specific host pathogen interactions by simply delivering R and Avr transgenes into the same cell. However, some plant genotypes can raise nonspecific defense responses to Agrobacterium, as we observed for example for several potato genotypes. Compared to agroinfiltration, detection of AVR activity with PVX agroinfection is more sensitive, more high-throughput in functional screens and less sensitive to nonspecific defense responses to Agrobacterium. However, nonspecific defense to PVX can occur and there is a risk to miss responses due to virus-induced extreme resistance. Despite such limitations, in our experience, agroinfiltration and PVX agroinfection are both suitable and complementary assays that can be used simultaneously to confirm each other's results.

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Section: Introductionmentioning

confidence: 99%

Agroinfiltration and PVX Agroinfection in Potato and <em>Nicotiana benthamiana</em>

Rietman

Vleeshouwers

2014

JoVE

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“…Predicting that genes encoding proteins necessary for the establishment of haustoria are under transcriptional control, we sequenced the whole haustorial transcriptome (Loehrer and Schaffrath, unpublished). Finally, we ended up with 111,440 de-novo assembled contigs from which 50 candidate genes were chosen based on structural features commonly found within known or predicted fungal effector genes (Ellis et al, 2007;Ellis et al, 2009). The functional testing of our candidate genes for an involvement in haustorium initiation requires the transformation of the pathogen.…”

Section: Analysis Of the Fungal-plant Interactome To Unravel P Pachymentioning

confidence: 99%

Asian Soybean Rust – Meet a Prominent Challenge in Soybean Cultivation

Loehrer¹,

Schaffrath²

2011

Soybean - Biochemistry, Chemistry and Physiology

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“…These include the RXLR motif that is required for translocation of secreted oomycete effectors inside the host cell cytoplasm, and appears to identify those effectors directed to host intracellular targets. 2,5,7,8,10,12 As a result, the oomycete effectors are currently grouped under different classes based on their modulatory activity toward plant defense and immunity. 6 Furthermore, the oomycete effector secretome comprises several uncharacterized proteins resulting from large-scale comparative genomics efforts aimed at elucidating the molecular mechanisms of Phytophthora pathogenicity.…”

Section: Introductionmentioning

confidence: 99%

Comparative structural and functional characterization of putative protein effectors belonging to the PcF toxin family from Phytophthora spp

et al. 2011

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The PcF Toxin Family (Pfam 09461) includes the characterized phytotoxic protein PcF from Phytophthora cactorum, as well as several predicted protein effectors from other Phytophthora species recently identified by comparative genomics. Here we provide first evidence that such 'putatives', recombinantly expressed in bacteria and purified to homogeneity, similarly to PcF, can trigger defense-related responses on tomato, that is leaf withering and phenylalanine ammonia lyase induction, although with various degrees of effectiveness. In addition, structural prediction by computer-aided homology modeling and subsequent structural/functional comparison after rational engineering of the disulfide-structured protein fold by site-directed mutagenesis, highlighted the surface-exposed conserved amino acid stretch SK(E/C)C as a possible structural determinant responsible for the differential phytotoxicity within this family of cognate protein effectors.

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Recent progress in discovery and functional analysis of effector proteins of fungal and oomycete plant pathogens

Cited by 152 publications

References 41 publications

Agroinfiltration and PVX Agroinfection in Potato and <em>Nicotiana benthamiana</em>

Agroinfiltration and PVX Agroinfection in Potato and <em>Nicotiana benthamiana</em>

Asian Soybean Rust – Meet a Prominent Challenge in Soybean Cultivation

Comparative structural and functional characterization of putative protein effectors belonging to the PcF toxin family from Phytophthora spp

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