Page 185 - Plant Canada 2024 Proceeding

PLANT CANADA 2024

finding applications for underutilized prairie berries as potential sources of functional food in the
pharmaceutical and nutraceutical industries.
Keywords: Bioactive compounds, UHPLC-HRMS, GC-MS, Canadian small fruits, antioxidants

*[O159] EXOGENOUS APPLICATIONS OF DOUBLE-STRANDED RNA TO INDUCE RNA
INTERFERENCE FOR THE CONTROL OF THE NOVEL FUNGAL PATHOGEN
1
1
NEOPESTALOTIOPSIS SP. AFFECTING STRAWBERRY. Sarah Koeppe and Melanie Kalischuk .
1 Department of Plant Agriculture, University of Guelph, 50 Stone Road East, Guelph, ON N1G 2W1,
Canada
Correspondence to: skoeppe@uoguelph.ca

Neopestalotiopsis sp. is an aggressive novel fungal pathogen of strawberries that currently lacks a
recommended control method. RNA interference (RNAi) is a mechanism triggered by double-stranded
RNA (dsRNA) that has the potential to be a highly effective and specific crop protection strategy,
however, its potential has yet to be explored for this pathogen. The RNA-induced silencing complex
(RISC) is a versatile, sequence-specific innate immunity protein complex involved in the RNAi pathway.
Exogenous application of dsRNA complementary to an integral gene of pathogen or pest can provide
systemic, RNAi-based resistance for the host crop in the form of spray-induced gene silencing (SIGS).
Two genes of interest that play integral roles in the functioning of this pathogen are beta-tubulin and
cytochrome P450 subfamily 51 (CYP51), both of which are targets for several fungicides. Beta-tubulin
plays an integral role in the formation of tubulin fibres during chromosome separation in cell division.
CYP51 has two paralogues in Neopestalotiopsis sp., and both encode for an enzyme that is necessary
for the production of ergosterol, a fungi-specific sterol needed to maintain cell membrane integrity. To
assess the efficacy of these genes as a target, they were first characterized to identify the locations of
introns, exons, splicing sites, and motifs. From this information, several dsRNA constructs were built, four
at the lengths of 200 base pairs (bp), 400bp, 600bp, and 800bp for beta-tubulin, and four for CYP51,
400bp and 800bp for each paralogue. An in vitro assay was developed to assess the efficacy of each
construct with their respective pathogen. Beta-tubulin showed no efficacy as an RNAi target for control of
Neopestalotiopsis sp., regardless of the construct used, potentially due to a resistance mechanism in the
pathogen. The CYP51 paralogues show some potential in controlling the pathogen but have yet to be
fully assessed. Based on the current research and these results, SIGS, and RNAi as a whole, appears to
be a potential environmentally friendly option for crop protection against fungal pathogens, and further
research in the field will allow for the development of alternative controls to conventional chemical
fungicides.

[O160] LEAFY GREEN VEGETABLE PRODUCTION IN SASKATCEWAN. Jazeem Wahab, Janitha
Wanasundara, Edmund Mupondwa, Erl Svendsen, Raju Soolanayakanahally, and Evan Derdall.
Saskatoon Research and Development Centre, Agriculture and Agri-Food Canada, 107 Science Place,
Saskatoon, SK S7N 0X2
Correspondence to: Jazeem.wahab@agr.gc.ca

Vegetable production is a leading sector of the Canadian Horticulture industry. In 2022, Canada grew
approximately 109,000 ha ‘Field’ vegetables with a Farm gate value of $ 1.6 billion. However,
Saskatchewan accounted for only about 0.5% of production area and 0.5% of Farm gate value. The
relatively short and cooler growing season is a major constraint for growing high-value warm-season
vegetables in Saskatchewan. Climate change mediated warmer temperatures and longer growing
seasons could be effectively capitalized to grow vegetables in Saskatchewan. Irrigation is essential for
sustainable vegetable production. Government of Saskatchewan’s recent 10-year - $ 4 billion - 202,000
ha irrigation expansion project expects vegetable production to play a key role. Many cool-season crops,
including Leafy-Greens’ have gained prominence as ‘Super Food’ or ‘Super Green’. Research is being
carried out at the Canada-Saskatchewan Irrigation Diversification Centre, Outlook, Saskatchewan, to
identify superior cultivars and develop Best Management Practices for popular ‘Greens’. Results from the
2022 growing season Bok Choy and kale studies are presented in this paper. Bok Choy cvs. Asian
Delight, Li Ren Choi, Mei Qing Choi, Win Win Choi, Bopak, and Joi Choi; and kale cvs. Starbor, White
Russian, Winterbor, Redbor, and Black Magic were evaluated at two sequential plantings under open-
field and high-tunnel production systems. The second planting was done about one week after harvesting
the first crop. The ‘Regular-type’ Bok Choy’ cv. Joi Choi produced the highest marketable yield under both

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