Apoplastic Sugar Extraction and Quantification from Wheat Leaves Infected with Biotrophic Fungi

Date

2017

Authors

Roman-Reyna, Veronica
Rathjen, John

Journal Title

Journal ISSN

Volume Title

Publisher

Springer Science+Business Media

Abstract

Biotrophic fungi such as rusts modify the nutrient status of their hosts by extracting sugars. Hemibiotrophic and biotrophic fungi obtain nutrients from the cytoplasm of host cells and/or the apoplastic spaces. Uptake of nutrients from the cytoplasm is via intracellular hyphae or more complex structures such as haustoria. Apoplastic nutrients are taken up by intercellular hyphae. Overall the infection creates a sink causing remobilization of nutrients from local and distal tissues. The main mobile sugar in plants is sucrose which is absorbed via plant or fungal transporters once unloaded into the cytoplasm or the apoplast. Infection by fungal pathogens alters the apoplastic sugar contents and stimulates the influx of nutrients towards the site of infection as the host tissue transitions to sink. Quantification of solutes in the apoplast can help to understand the allocation of nutrients during infection. However, separation of apoplastic fluids from whole tissue is not straightforward and leakage from damaged cells can alter the results of the extraction. Here, we describe how variation in cytoplasmic contamination and infiltrated leaf volumes must be controlled when extracting apoplastic fluids from healthy and rust-infected wheat leaves. We show the importance of correcting the data for these parameters to measure sugar concentrations accurately.

Description

Keywords

Rust fungi, Puccinia striiformis f. sp. tritici, Apoplastic fluids, Hexoses, Cytoplasmic contamination

Citation

Source

Type

Book chapter

Book Title

Wheat Rust Diseases: Methods and Protocols

Entity type

Access Statement

License Rights

Restricted until

2099-12-31