Deploying calcium-dependent protein kinases to fight canola pathogens

Project Summary

Overview

Agriculture is one of Canada’s most important industries. Grown outside and in greenhouses, our crops are susceptible to dynamic environmental conditions such as fluctuating temperature, moisture, light, and exposure to a number of pests and pathogens. Fungi, oomycetes, viruses, bacteria, nematodes, and chewing insects wreak havoc on our crops, and this is expected to worsen with global climate change. Even in the absence of significant outbreaks, diseases such as clubroot and stem rot of canola account for ~25% of crop and yield losses annually.

Current efforts to mitigate crop damage typically rely on chemical inputs (i.e., pesticides), which can present unfavourable trade-offs to the larger ecosystem. As there continues to be no evidence that genetically modified plants used for food production are harmful to humans, livestock, or the environment, the most sustainable solution to these problems is the development of elite plant varieties that are resistant to pathogen infection.

Enhancing pathogen resistance is particularly challenging for two reasons: (a) Similar to the flu, new pathotypes continually arise that can overcome plant immunity, and (b) Similar to autoimmune diseases, plant immune signaling must be finely tuned to avoid self-damage.

Monaghan’s research team proposes using precision gene editing to make single base pair changes in the canola genome to enhance the function of single genes that are poised to result in plants with enhanced, durable, broad-spectrum resistance without any growth tradeoff.

Purpose

This project will contribute to the development of elite canola varieties that are resistant to pathogen infection for the betterment of the canola industry. As a result, this research will have a great impact at different levels:

(a) In our research environments, through training of five undergraduate students, one Master student, and a Postdoctoral Scholar, and through the opportunity to expand our research in canola

(b) For canola growers and the canola industry in Canada and elsewhere, because the results obtained through this project works towards deploying precision-engineered genetic loci that are likely to result in broad-spectrum resistance

(c) For our society, because stable and robust canola resistance will allow economic stability and a healthy environment, taking into account that aside from genetic resistance, current methods to manage clubroot and stem rot rely on the application of high amounts of chemicals.

From the local perspective, this project will also have a lasting impact by working closely with provincial governments and canola specialists (through existing and newly established partnerships) and with the Manitoba Canola Growers Association, SaskCanola, and Alberta Canola to ensure the effective transfer of the results.

Objectives

The objectives of this project are:

Objective 1. Biochemical and functional assessment of subgroup IV CPKs in canola (Years 1-3)
● 1a. Select candidate BnCPKs.
● 1b. Identify catalytically active candidate BnCPKs.
● 1c. Confirm membrane localization of candidate BnCPKs.
● 1d. Assess the function of BnCPK variants using Arabidopsis as a surrogate.
Objective 2. Precision-engineer BnCPK alleles for enhanced disease resistance in canola (Years 3-5)
● 2a. Base-edit BnCPKs directly in canola.
● 2b. Assess the effect of base-edited BnCPKs variants in canola development.
● 2c. Assess the effect of base-edited BnCPKs variants in canola disease resistance.

References