Alberta Barley

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Feed and malting barley: linking enzymes to disease resistance

Project lead:
Dr. Jennifer Zantinge
Research Scientist, Molecular Biology
Alberta Agriculture and Forestry

Timeline: April 2014 – March 2017

Partners:
AB-AAFC-BMBRI-WGRF

Alberta Barley funding: $20,000
Total funding from other partners: $202,000

Benefits for barley farmers
The findings of this project will lead to more durable resistance in elite barley varieties to barley diseases FHB, net blotch and scald. In vitro assays that identify the plant reaction to disease will improve the quality of disease screening by better characterizing the disease resistance mechanism. Improved phenotyping methods for disease resistance may be directly applied within the breeding program or used to improve molecular marker selection.

Summary
We plan to develop in vitro or lab-based phenotyping methods that accurately characterize disease resistance mechanisms and that can be used for quick disease resistance selection with in a breeding program.  This will be done by developing lab based methods to assay pathogen-related enzymes (glucanases, chitiases, peroxidases, etc.) linked to disease resistant or susceptible phenotype.

Recent developments
Peroxides are frequently responsive to early cell signaling resulting from pathogen recognition. Higher expression of peroxidase activity has been linked to increased fungal disease resistance in plants. Preliminary results suggest peroxidase assay (DAB assay) has potential for use within the FCDC breeding program and further evaluation of the DAB assay will continue in 2016-17.

Objectives for the upcoming year
We will compare the current leaf assay (DAB assay) with easier biochemical-based assays.  Enzyme expression in the barley secretome after pathogen inoculation will also be further characterized. Enzymes that differ between resistant and susceptible disease barley-pathogen reactions, will be further evaluated as targets for new in vitro assays.

P80 chip run of AWF fluid from leaves collected at 5 dpi with SFNB (Ptm14) and NFNB (WG17) from barley varieties Harrington (Har), CDC Dolly (Dolly), CDC Helgason and Vivar. Control samples are marked with a “C”, and “R” is for rep sample. The gel-like image representation of AWF protein are shown in the figure after alignment with lower marker (green) and upper marker (purple). The protein Ladder is labeled on the side of image. Some of the bands that vary between samples are marked with an arrow. Red arrows show an increase in susceptible varieties will black arrows appear to increase in resistant varieties.

P80 chip run of AWF fluid from leaves collected at 5 dpi with SFNB (Ptm14) and NFNB (WG17) from barley varieties Harrington (Har), CDC Dolly (Dolly), CDC Helgason and Vivar. Control samples are marked with a “C”, and “R” is for rep sample. The gel-like image representation of AWF protein are shown in the figure after alignment with lower marker (green) and upper marker (purple). The protein Ladder is labeled on the side of image. Some of the bands that vary between samples are marked with an arrow. Red arrows show an increase in susceptible varieties will black arrows appear to increase in resistant varieties.