Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses

Using resistant cultivars is a common approach to managing blackleg of canola/rapeseed caused by <i>Leptosphaeria maculans</i> (<i>Lm</i>). Quantitative resistance (QR), as opposed to major-gene resistance, is of interest because it is generally more durable, due to its multi...

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Main Authors: Michelle Hubbard, Chun Zhai, Gary Peng
Format: Article
Language:English
Published: MDPI AG 2020-07-01
Series:Plants
Subjects:
Online Access:https://www.mdpi.com/2223-7747/9/7/864
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spelling doaj-d7f7f17807db49d2a82ff46895713e682020-11-25T03:22:08ZengMDPI AGPlants2223-77472020-07-01986486410.3390/plants9070864Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic AnalysesMichelle Hubbard0Chun Zhai1Gary Peng2Agriculture and Agri-Food Canada, Swift Current Research and Development Centre, Swift Current, Saskatchewan S9H 3X2, CanadaAgriculture and Agri-Food Canada, Saskatoon Research and Development Centre, Saskatoon, Saskatchewan S7N 0X2, CanadaAgriculture and Agri-Food Canada, Saskatoon Research and Development Centre, Saskatoon, Saskatchewan S7N 0X2, CanadaUsing resistant cultivars is a common approach to managing blackleg of canola/rapeseed caused by <i>Leptosphaeria maculans</i> (<i>Lm</i>). Quantitative resistance (QR), as opposed to major-gene resistance, is of interest because it is generally more durable, due to its multi-genetic basis. However, the mechanisms and genes underlying QR are mostly unknown. In this study, potential QR modes of action in “74‑44 BL” was explored. This Canadian canola cultivar showed moderate but consistent race-nonspecific resistance at the cotyledon and adult-plant stages. A susceptible cultivar, “Westar”, was used as a control. After inoculation, the lesions developed more slowly on the cotyledons of 74-44 BL than those of Westar. We used RNA sequencing (-RNA-seq) to identify genes and their functions, putatively related to this resistance, and found that genes involved in programmed cell death (PCD), reactive oxygen species (ROS), signal transduction or intracellular endomembrane transport were most differentially expressed. ROS production was assessed in relation to <i>Lm</i> hyphal growth and lesion size; it occurred beyond the tissue colonized by <i>Lm</i> in 74-44 BL and appeared to trigger rapid cell death, limiting cotyledon colonization by <i>Lm</i>. In contrast, <i>Lm</i> grew more rapidly in Westar, often catching up with the ring of ROS and surpassing lesion boundaries. It appears that QR in 74-44 BL cotyledons is associated with limited colonization by <i>Lm </i>possibly mediated via ROS. The RNA-seq data also showed a link between ROS, signal transduction, and endomembrane vesicle trafficking, as well as PCD in the resistance. These results provide a starting point for a better understanding of the mechanisms behind QR against <i>Lm</i> in canola.https://www.mdpi.com/2223-7747/9/7/864<i>Leptosphaeria maculans</i>canolaPhoma stem cankergenetic resistancetranscriptome analysisRNA-seq
collection DOAJ
language English
format Article
sources DOAJ
author Michelle Hubbard
Chun Zhai
Gary Peng
spellingShingle Michelle Hubbard
Chun Zhai
Gary Peng
Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
Plants
<i>Leptosphaeria maculans</i>
canola
Phoma stem canker
genetic resistance
transcriptome analysis
RNA-seq
author_facet Michelle Hubbard
Chun Zhai
Gary Peng
author_sort Michelle Hubbard
title Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
title_short Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
title_full Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
title_fullStr Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
title_full_unstemmed Exploring Mechanisms of Quantitative Resistance to <i>Leptosphaeria maculans</i> (Blackleg) in the Cotyledons of Canola (<i>Brassica napus</i>) Based on Transcriptomic and Microscopic Analyses
title_sort exploring mechanisms of quantitative resistance to <i>leptosphaeria maculans</i> (blackleg) in the cotyledons of canola (<i>brassica napus</i>) based on transcriptomic and microscopic analyses
publisher MDPI AG
series Plants
issn 2223-7747
publishDate 2020-07-01
description Using resistant cultivars is a common approach to managing blackleg of canola/rapeseed caused by <i>Leptosphaeria maculans</i> (<i>Lm</i>). Quantitative resistance (QR), as opposed to major-gene resistance, is of interest because it is generally more durable, due to its multi-genetic basis. However, the mechanisms and genes underlying QR are mostly unknown. In this study, potential QR modes of action in “74‑44 BL” was explored. This Canadian canola cultivar showed moderate but consistent race-nonspecific resistance at the cotyledon and adult-plant stages. A susceptible cultivar, “Westar”, was used as a control. After inoculation, the lesions developed more slowly on the cotyledons of 74-44 BL than those of Westar. We used RNA sequencing (-RNA-seq) to identify genes and their functions, putatively related to this resistance, and found that genes involved in programmed cell death (PCD), reactive oxygen species (ROS), signal transduction or intracellular endomembrane transport were most differentially expressed. ROS production was assessed in relation to <i>Lm</i> hyphal growth and lesion size; it occurred beyond the tissue colonized by <i>Lm</i> in 74-44 BL and appeared to trigger rapid cell death, limiting cotyledon colonization by <i>Lm</i>. In contrast, <i>Lm</i> grew more rapidly in Westar, often catching up with the ring of ROS and surpassing lesion boundaries. It appears that QR in 74-44 BL cotyledons is associated with limited colonization by <i>Lm </i>possibly mediated via ROS. The RNA-seq data also showed a link between ROS, signal transduction, and endomembrane vesicle trafficking, as well as PCD in the resistance. These results provide a starting point for a better understanding of the mechanisms behind QR against <i>Lm</i> in canola.
topic <i>Leptosphaeria maculans</i>
canola
Phoma stem canker
genetic resistance
transcriptome analysis
RNA-seq
url https://www.mdpi.com/2223-7747/9/7/864
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