Zur Kultur von <i>Phytophthora infestans</i> auf vollsynthetischen Nährsubstraten

Henniger, H.

doi:10.1002/jobm.19630030204

Cited by 13 publications

(17 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Identification of a 15 kDa glycoprotein from culture filtrate of P. palmivora. To identify potential elicitors, culture filtrate of P. palmivora grown in Henniger medium 32 was collected, dialyzed and lyophilized, and then separated on 15% SDS-PAGE. Two replicative gels were stained with InstantBlue Coomassie Protein Stain (Expedeon) and periodic acid-Schiff reagent for visualization of total proteins and glycoproteins, respectively.…”

Section: Resultsmentioning

confidence: 99%

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity

Pettongkhao

Navet

Schornack

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Phytophthora palmivora is a destructive oomycete plant pathogen with a wide host range. So far, little is known about the factors governing its infection structure development and pathogenicity. From the culture filtrate of a P. palmivora strain isolated from papaya, we identified a secreted glycoprotein of 15 kDa, designated as Ppal15kDa, using liquid chromatography tandem mass spectrometry. Two gene variants, Ppal15kDaA and Ppal15kDaB were amplified from a P. palmivora papaya isolate. Transient expression of both variants in Nicotiana benthamiana by agroinfiltration enhanced P. palmivora infection. Six Ppal15kDa mutants with diverse mutations were generated via CRISPR/Cas9-mediated gene editing. All mutants were compromised in infectivity on N. benthamiana and papaya. Two mutants with all Ppal15kDa copies mutated almost completely lost pathogenicity. The pathogenicity of the other four containing at least one wild-type copy of Ppal15kDa was compromised at varying levels. The mutants were also affected in development as they produced smaller sporangia, shorter germ tubes, and fewer appressoria. The affected levels in development corresponded to the levels of reduction in pathogenicity, suggesting that Ppal15kDa plays an important role in normal development of P. palmivora infection structures. Consistent with its role in infection structure development and pathogenicity, Ppal15kDa was found to be highly induced during appressorium formation. In addition, Ppal15kDa homologs are broadly present in Phytophthora spp., but none were characterized. Altogether, this study identified a novel component involved in development and pathogenicity of P. palmivora and possibly other Phytophthora spp. known to contain a Ppal15kDa homolog. Oomycetes are fungal-like microorganisms belonging to the kingdom Straminipila 1,2. Their characteristics are distinct from true fungi since oomycetes are diploid or polyploid whereas fungi are haploid for most of the life cycles 2-4. In addition, cell walls of oomycetes mainly consist of 1,3-beta-glucan, while the major cell wall constituent of fungi is chitin 5. Oomycetes include many destructive plant pathogens, among which are over 100 species in the genus Phytophthora that severely threaten agricultural production and natural ecosystems 5,6. Phytophthora palmivora is a hemibiotrophic oomycete pathogen that infects more than 200 plant species in the tropics and subtropics 5. Examples of economically important hosts include papaya, cacao, pineapple, durian, rubber tree, citrus, and oil palm. It also infects model plant species, such as Nicotiana benthamiana and Medicago truncatula 7-9. Similar to other Phytophthora spp., plant infection by P. palmivora starts with motile zoospores, which encyst after contacting plant surfaces, followed by formation of germ tubes and then appressoria to penetrate the plant surface 7,10,11. During infection, P. palmivora initially grows as a biotroph by forming haustoria inside the host cells to obtain nutrients, and then switches to necrotrophy in the later...

show abstract

Section: Resultsmentioning

confidence: 99%

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity

Pettongkhao

Navet

Schornack

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…However, their study used a different medium, the amino acid-based Henninger medium [44]. We therefore performed the experiment using that medium (Fig 8B).…”

Section: Resultsmentioning

confidence: 99%

“…Cultures were maintained in the dark on rye-sucrose agar [31] at 18°C. Conditions for RNA analysis employed rye-sucrose agar, a defined minimal medium based on the recipe of Xu [32], the latter with (NH 4 ) 2 SO 4 omitted and replaced by 1% casamino acids, or Henninger medium [44]. Some cultures were amended with KNO 3 or (NH 4 ) 2 SO 4 as described in Results.…”

Section: Methodsmentioning

confidence: 99%

Gene Expression and Silencing Studies in Phytophthora infestans Reveal Infection-Specific Nutrient Transporters and a Role for the Nitrate Reductase Pathway in Plant Pathogenesis

et al. 2016

View full text Add to dashboard Cite

To help learn how phytopathogens feed from their hosts, genes for nutrient transporters from the hemibiotrophic potato and tomato pest Phytophthora infestans were annotated. This identified 453 genes from 19 families. Comparisons with a necrotrophic oomycete, Pythium ultimum var. ultimum, and a hemibiotrophic fungus, Magnaporthe oryzae, revealed diversity in the size of some families although a similar fraction of genes encoded transporters. RNA-seq of infected potato tubers, tomato leaves, and several artificial media revealed that 56 and 207 transporters from P. infestans were significantly up- or down-regulated, respectively, during early infection timepoints of leaves or tubers versus media. About 17 were up-regulated >4-fold in both leaves and tubers compared to media and expressed primarily in the biotrophic stage. The transcription pattern of many genes was host-organ specific. For example, the mRNA level of a nitrate transporter (NRT) was about 100-fold higher during mid-infection in leaves, which are nitrate-rich, than in tubers and three types of artificial media, which are nitrate-poor. The NRT gene is physically linked with genes encoding nitrate reductase (NR) and nitrite reductase (NiR), which mobilize nitrate into ammonium and amino acids. All three genes were coregulated. For example, the three genes were expressed primarily at mid-stage infection timepoints in both potato and tomato leaves, but showed little expression in potato tubers. Transformants down-regulated for all three genes were generated by DNA-directed RNAi, with silencing spreading from the NR target to the flanking NRT and NiR genes. The silenced strains were nonpathogenic on leaves but colonized tubers. We propose that the nitrate assimilation genes play roles both in obtaining nitrogen for amino acid biosynthesis and protecting P. infestans from natural or fertilization-induced nitrate and nitrite toxicity.

show abstract

“…Candidate target genes were selected based on the stability of their CPM (counts per million) values in nonsporulating hyphae grown in pea broth, rye broth, rye agar, amino acid-based minimal media [ 31 ], ammonium sulfate-based minimal media [ 32 ], sporulating hyphae on rye agar, 10-day matings on rye agar, hyphae treated with metalaxyl in rye broth (6 hr at IC 10 ), sporangia, sporangia chilled to stimulate zoosporogenesis, swimming zoospores, and germinated cysts forming appressoria [ 33 ], and infected potato tubers and tomato leaves at early, middle, and late stages of infection [ 34 , 35 ]. Primers ( S1 Table ) were designed to amplify bands of about 150 bp from the 3’ end of the target mRNA.…”

Section: Methodsmentioning

confidence: 99%

Transcriptomic and proteomic analysis reveals wall-associated and glucan-degrading proteins with potential roles in Phytophthora infestans sexual spore development

et al. 2018

View full text Add to dashboard Cite

Sexual reproduction remains an understudied feature of oomycete biology. To expand our knowledge of this process, we used RNA-seq and quantitative proteomics to examine matings in Phytophthora infestans. Exhibiting significant changes in mRNA abundance in three matings between different A1 and A2 strains compared to nonmating controls were 1170 genes, most being mating-induced. Rising by >10-fold in at least one cross were 455 genes, and 182 in all three crosses. Most genes had elevated expression in a self-fertile strain. Many mating-induced genes were associated with cell wall biosynthesis, which may relate to forming the thick-walled sexual spore (oospore). Several gene families were induced during mating including one encoding histidine, serine, and tyrosine-rich putative wall proteins, and another encoding prolyl hydroxylases which may strengthen the extracellular matrix. The sizes of these families vary >10-fold between Phytophthora species and one exhibits concerted evolution, highlighting two features of genome dynamics within the genus. Proteomic analyses of mature oospores and nonmating hyphae using isobaric tags for quantification identified 835 shared proteins, with 5% showing >2-fold changes in abundance between the tissues. Enriched in oospores were β-glucanases potentially involved in digesting the oospore wall during germination. Despite being dormant, oospores contained a mostly normal complement of proteins required for core cellular functions. The RNA-seq data generated here and in prior studies were used to identify new housekeeping controls for gene expression studies that are more stable than existing normalization standards. We also observed >2-fold variation in the fraction of polyA+ RNA between life stages, which should be considered when quantifying transcripts and may also be relevant to understanding translational control during development.

show abstract

Zur Kultur von Phytophthora infestans auf vollsynthetischen Nährsubstraten

Cited by 13 publications

References 9 publications

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity

Gene Expression and Silencing Studies in Phytophthora infestans Reveal Infection-Specific Nutrient Transporters and a Role for the Nitrate Reductase Pathway in Plant Pathogenesis

Transcriptomic and proteomic analysis reveals wall-associated and glucan-degrading proteins with potential roles in Phytophthora infestans sexual spore development

Contact Info

Product

Resources

About