Evidence that PSI-D, a chloroplast photosystem I protein, is in haustoria of the powdery mildew fungus Erysiphe pisi

Testut, J.-F.; Callow, James A.; Green, J.R.

doi:10.1006/pmpp.1999.0238

Cited by 2 publications

(2 citation statements)

References 24 publications

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“…In contrast to the membranes and organelles in very close proximity to the EHM on the side of the host (Kwaaitaal et al, ), transmission electron micrographs have shown the EHMx to be easily discernible and up to 3 μm wide (Hippe‐Sanwald, Hermanns, & Somerville, ). Interestingly, the chloroplast protein, PSI‐D, was localised in haustoria of Erysiphe pisi growing on pea, thus suggesting protein transport across both the EHM and the fungal plasma membrane (Testut, Callow, & Green, ). Otherwise, molecular and cellular information available on the haustorial complex is very sparse, despite the obvious importance of this compartment.…”

Section: Introductionmentioning

confidence: 99%

The isoelectric point of proteins influences their translocation to the extrahaustorial matrix of the barley powdery mildew fungus

Smigielski

Aguilar

Kwaaitaal

et al. 2019

Cellular Microbiology

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Many biotrophic fungal plant pathogens develop feeding structures, haustoria, inside living plant cells, which are essential for their success. Extrahaustorial membranes (EHMs) surround haustoria and delimit the extrahaustorial matrices (EHMxs). Little is known about transport mechanisms across EHMs and what properties proteins and nutrients need in order to cross these membranes. To investigate this further, we expressed fluorescent proteins in the cytosol of infected barley leaf epidermal cells after particle bombardment and investigated properties that influenced their localisation in the powdery mildew EHMx. We showed that this translocation is favoured by a neutral isoelectric point (pI) between 6.0 and 8.4. However, for proteins larger than 50 kDa, pI alone does not explain their localisation, hinting towards a more complex interplay between pI, size, and sequence properties. We discuss the possibility that an EHM translocon is involved in protein uptake into the EHMx.

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Section: Introductionmentioning

confidence: 99%

The isoelectric point of proteins influences their translocation to the extrahaustorial matrix of the barley powdery mildew fungus

Smigielski

Aguilar

Kwaaitaal

et al. 2019

Cellular Microbiology

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“…Previously, the procedure for isolating haustoria from obligate biotrophs employed affinity chromatography on rust fungi (Catanzariti et al, 2006;Garnica et al, 2014). During the 1990s, several centrifugation-based methods were introduced to isolate powdery mildew haustoria from pea plants (Mackie et al, 1991;Testut et al, 1999). This method was then applied to powdery mildews of other plants, including Arabidopsis (Wang et al, 2009;Micali et al, 2011).…”

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Isolation of Powdery Mildew Haustoria from Infected Barley

Li¹,

Collier²,

Spanu³

2019

BIO-PROTOCOL

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Blumeria graminis is a fungus that causes powdery mildews on grasses, such as barley.Investigations of this pathogen present many challenges due to its obligate biotrophic nature. This means that the fungus can only grow in the presence of a living host plant. B. graminis forms epiphytic mycelia on the plant surface and feeding organs (haustoria) inside the epidermal cells of the host plant.Therefore, it is difficult to separate the fungus from plant tissues. This protocol shows how to obtain different fungal structures from powdery mildew infected barley leaves. The epiphytic mycelia including conidia and conidiophores can be separated after immersing the infected leaves into 5% cellulose acetate dissolved in acetone, and peeling off the cellulose acetate membrane. Then, the haustoria are isolated from dissected epidermis after cellulase degradation of plant cell walls. The isolated haustoria remain intact with few plant impurities. The haustoria may be visualized by epifluorescence microscopy after staining with the chitin-specific dye WGA-Alexa Fluor 488. Finally, dissected material can be either processed immediately or kept at -80 °C for long-term storage for studies on gene expression and protein identification, for example by mass spectrometry.

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Evidence that PSI-D, a chloroplast photosystem I protein, is in haustoria of the powdery mildew fungus Erysiphe pisi

Cited by 2 publications

References 24 publications

The isoelectric point of proteins influences their translocation to the extrahaustorial matrix of the barley powdery mildew fungus

The isoelectric point of proteins influences their translocation to the extrahaustorial matrix of the barley powdery mildew fungus

Isolation of Powdery Mildew Haustoria from Infected Barley

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