Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction

Abstract Background Blackleg disease, caused by the fungal pathogen Leptosphaeria maculans, is a serious threat to canola (Brassica napus) production worldwide. Quantitative resistance to this disease is a highly desirable trait but is difficult to precisely phenotype. Visual scores can be subjectiv...

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Autores principales:	Wendelin Schnippenkoetter, Mohammad Hoque, Rebecca Maher, Angela Van de Wouw, Phillip Hands, Vivien Rolland, Luke Barrett, Susan Sprague
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Canola (Brassica napus) Blackleg (Leptosphaeria maculans) Disease resistance Pathogen Fungal biomass assay Phenotyping Plant culture SB1-1110 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/5b06382e9e494de7b52b4fd3ca1084fb
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spelling	oai:doaj.org-article:5b06382e9e494de7b52b4fd3ca1084fb2021-12-05T12:07:43ZComparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction10.1186/s13007-021-00822-61746-4811https://doaj.org/article/5b06382e9e494de7b52b4fd3ca1084fb2021-12-01T00:00:00Zhttps://doi.org/10.1186/s13007-021-00822-6https://doaj.org/toc/1746-4811Abstract Background Blackleg disease, caused by the fungal pathogen Leptosphaeria maculans, is a serious threat to canola (Brassica napus) production worldwide. Quantitative resistance to this disease is a highly desirable trait but is difficult to precisely phenotype. Visual scores can be subjective and are prone to assessor bias. Methods to assess variation in quantitative resistance more accurately were developed based on quantifying in planta fungal biomass, including the Wheat Germ Agglutinin Chitin Assay (WAC), qPCR and ddPCR assays. Results Disease assays were conducted by inoculating a range of canola cultivars with L. maculans isolates in glasshouse experiments and assessing fungal biomass in cotyledons, petioles and stem tissue harvested at different timepoints post-inoculation. PCR and WAC assay results were well correlated, repeatable across experiments and host tissues, and able to differentiate fungal biomass in different host-isolate treatments. In addition, the ddPCR assay was shown to differentiate between L. maculans isolates. Conclusions The ddPCR assay is more sensitive in detecting pathogens and more adaptable to high-throughput methods by using robotic systems than the WAC assay. Overall, these methods proved accurate and non-subjective, providing alternatives to visual assessments to quantify the L. maculans-B. napus interaction in all plant tissues throughout the progression of the disease in seedlings and mature plants and have potential for fine-scale blackleg resistance phenotyping in canola.Wendelin SchnippenkoetterMohammad HoqueRebecca MaherAngela Van de WouwPhillip HandsVivien RollandLuke BarrettSusan SpragueBMCarticleCanola (Brassica napus)Blackleg (Leptosphaeria maculans)Disease resistancePathogenFungal biomass assayPhenotypingPlant cultureSB1-1110Biology (General)QH301-705.5ENPlant Methods, Vol 17, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Canola (Brassica napus) Blackleg (Leptosphaeria maculans) Disease resistance Pathogen Fungal biomass assay Phenotyping Plant culture SB1-1110 Biology (General) QH301-705.5
spellingShingle	Canola (Brassica napus) Blackleg (Leptosphaeria maculans) Disease resistance Pathogen Fungal biomass assay Phenotyping Plant culture SB1-1110 Biology (General) QH301-705.5 Wendelin Schnippenkoetter Mohammad Hoque Rebecca Maher Angela Van de Wouw Phillip Hands Vivien Rolland Luke Barrett Susan Sprague Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
description	Abstract Background Blackleg disease, caused by the fungal pathogen Leptosphaeria maculans, is a serious threat to canola (Brassica napus) production worldwide. Quantitative resistance to this disease is a highly desirable trait but is difficult to precisely phenotype. Visual scores can be subjective and are prone to assessor bias. Methods to assess variation in quantitative resistance more accurately were developed based on quantifying in planta fungal biomass, including the Wheat Germ Agglutinin Chitin Assay (WAC), qPCR and ddPCR assays. Results Disease assays were conducted by inoculating a range of canola cultivars with L. maculans isolates in glasshouse experiments and assessing fungal biomass in cotyledons, petioles and stem tissue harvested at different timepoints post-inoculation. PCR and WAC assay results were well correlated, repeatable across experiments and host tissues, and able to differentiate fungal biomass in different host-isolate treatments. In addition, the ddPCR assay was shown to differentiate between L. maculans isolates. Conclusions The ddPCR assay is more sensitive in detecting pathogens and more adaptable to high-throughput methods by using robotic systems than the WAC assay. Overall, these methods proved accurate and non-subjective, providing alternatives to visual assessments to quantify the L. maculans-B. napus interaction in all plant tissues throughout the progression of the disease in seedlings and mature plants and have potential for fine-scale blackleg resistance phenotyping in canola.
format	article
author	Wendelin Schnippenkoetter Mohammad Hoque Rebecca Maher Angela Van de Wouw Phillip Hands Vivien Rolland Luke Barrett Susan Sprague
author_facet	Wendelin Schnippenkoetter Mohammad Hoque Rebecca Maher Angela Van de Wouw Phillip Hands Vivien Rolland Luke Barrett Susan Sprague
author_sort	Wendelin Schnippenkoetter
title	Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
title_short	Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
title_full	Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
title_fullStr	Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
title_full_unstemmed	Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction
title_sort	comparison of non-subjective relative fungal biomass measurements to quantify the leptosphaeria maculans—brassica napus interaction
publisher	BMC
publishDate	2021
url	https://doaj.org/article/5b06382e9e494de7b52b4fd3ca1084fb
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Comparison of non-subjective relative fungal biomass measurements to quantify the Leptosphaeria maculans—Brassica napus interaction

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