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ARS Home » Pacific West Area » Pullman, Washington » Grain Legume Genetics Physiology Research » Research » Publications at this Location » Publication #404375
Research Project: Enhanced Agronomic Performance and Disease Resistance in Edible Legumes

Location: Grain Legume Genetics Physiology Research

Title: Automating High-throughput Screening for Anthracnose Resistance in Common Bean Using Allele Specific PCR

Author
item ZALESKI-COX, MARYSIA - McGill University - Canada
item Miklas, Phillip - Phil
item SOLER-GAARZON, ALVARO - Washington State University
item HOYES-VILLEGAS, VALERIO - McGill University - Canada

Submitted to: Plant Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/15/2023
Publication Date: 10/3/2023
Citation: Zaleski-Cox, M., Miklas, P.N., Soler-Gaarzon, A., Hoyes-Villegas, V. 2023. Automating High-throughput Screening for Anthracnose Resistance in Common Bean Using Allele Specific PCR. Plant Methods. 19:102. Https://doi.org/10.1186/s13007-023-01071-5.
DOI: https://doi.org/10.1186/s13007-023-01071-5

Interpretive Summary: This research was conducted to improve throughput for marker-assisted selection applications in common bean. A liquid handling robot (LHR) was used for DNA extraction and molecular marker assays for anthracnose-resistance linked markers in common bean. Anthracnose is an important fungal disease in North America which limits bean production. Resistance genes provide for the best control of the disease but screening for resistance genes using pathogen assays are cumbersome and time-consuming. This new robotic method appears to significantly reduce technical labor costs for molecular marker assays by about 25%. Further validation of this new technique is pending.

Technical Abstract: Common beans (Phaseolus vulgaris L.) provide important protein and calories globally. Anthracnose (Colletotrichum lindemuthianum (Sacc. & Magnus) Briosi & Cavara, 1889) is a major disease in common bean and causes significant yield losses in bean production areas. Screening for markers linked to known disease resistance genes provides useful information for plant breeders to develop improved common bean varieties. The Kompetitive Allele Specific PCR (KASP) assay is an affordable genetic screening technique that can be used to accelerate breeding programs, but manual DNA extraction and KASP assay preparation are time-consuming. Several KASP markers have been developed for genes involved in resistance to bean anthracnose, which can reduce yield by up to 100%, but their usefulness is hindered by the labor required to screen a significant number of bean lines. Our research objective was to develop publicly available protocols for DNA extraction and KASP assaying using a liquid handling robot (LHR) which would facilitate high-throughput genetic screening with less active human time required. Anthracnose resistance markers were used to compare manual and automated results.