Identification of Micro Ribonucleic Acids and Their Targets in Response to Plasmodiophora brassicae Infection in Brassica napus

Clubroot disease, which is caused by the soil-borne pathogen Plasmodiophora brassicae War (P. brassicae), is one of the oldest and most destructive diseases of Brassica and cruciferous crops in the world. Plant microRNAs [micro ribonucleic acids (miRNAs)] play important regulatory roles in several developmental processes. Although the role of plant miRNAs in plant-microbe interaction has been extensively studied, there are only few reports on the specific functions of miRNAs in response to P. brassicae. This study investigated the roles of miRNAs and their targets during P. brassicae infection in a pair of Brassica napus near-isogenic lines (NILs), namely clubroot-resistant line 409R and clubroot-susceptible line 409S. Small RNA sequencing (sRNA-seq) and degradome-seq were performed on root samples of 409R and 409S with or without P. brassicae inoculation. sRNA-seq identified a total of 48 conserved and 72 novel miRNAs, among which 18 had a significant differential expression in the root of 409R, while only one miRNA was differentially expressed in the root of 409S after P. brassicae inoculation. The degradome-seq analysis identified 938 miRNA target transcripts, which are transcription factors, enzymes, and proteins involved in multiple biological processes and most significantly enriched in the plant hormone signal transduction pathway. Between 409R and 409S, we found eight different degradation pathways in response to P. brassicae infection, such as those related to fatty acids. By combining published transcriptome data, we identified a total of six antagonistic miRNA-target pairs in 409R that are responsive to P. brassicae infection and involved in pathways associated with root development, hypersensitive cell death, and chloroplast metabolic synthesis. Our results reveal that P. brassicae infection leads to great changes in miRNA pool and target transcripts. More interestingly, these changes are different between 409R and 409S. Clarification of the crosstalk between miRNAs and their targets may shed new light on the possible mechanisms underlying the pathogen resistance against P. brassicae.

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Suppression of Canola (Brassica napus) Resistance by Virulent Isolates of Plasmodiophora brassicae (Clubroot) During Primary Infection

Plant Disease ◽

10.1094/pdis-03-19-0659-re ◽

2020 ◽

Vol 104 (2) ◽

pp. 430-437

Author(s):

Junye Jiang ◽

Rudolph Fredua-Agyeman ◽

Stephen E. Strelkov ◽

Sheau-Fang Hwang

Keyword(s):

Brassica Napus ◽

Plasmodiophora Brassicae ◽

Negative Control ◽

Pcr Analysis ◽

High Concentration ◽

Clubroot Disease ◽

Low Concentrations ◽

High Concentrations ◽

Experimental Treatments ◽

Positive Controls

The planting of clubroot resistant (CR) canola (Brassica napus) is the most effective method to manage clubroot. Since 2013, many Plasmodiophora brassicae isolates capable of overcoming resistance have been detected, often in mixtures with avirulent isolates. To improve understanding of the effect of low concentrations of virulent isolates on host resistance, three CR canola cultivars (45H29, L135C, and L241C) were inoculated with pairs of isolates representing virulent/avirulent pathotypes (2*/2, 3*/3, and 5*/5) collected after or before the introduction of CR canola, respectively. Seven-day-old seedlings of each cultivar were incubated for 2 days in low concentrations (1 × 103 spores/ml) of the virulent isolates, followed by a second inoculation with a high concentration (1 × 107 spores/ml) of the avirulent isolates. Positive controls comprised seedlings inoculated with low concentrations of the virulent isolates followed by high concentrations of the virulent isolates (PC1) or only with high concentrations of virulent isolates (PC2). Negative controls comprised seedlings inoculated only with high concentrations of the avirulent isolates (NC1) or only with low concentrations of the virulent isolates (NC2). Clubroot severity was significantly higher in all nine experimental treatments (low virulent plus high avirulent) than in the negative control NC1 (high avirulent) but was lower in the experimental treatments than in the positive controls (PC1 and PC2). Low concentrations of virulent isolates alone (NC2) caused moderate clubroot. Disease severity correlated well with P. brassicae biomass in canola as determined by quantitative PCR analysis 28 to 35 days after inoculation. This study revealed that low concentrations of virulent isolates compromised canola resistance for infection by avirulent isolates.

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Genetics and molecular mapping of resistance to Plasmodiophora brassicae pathotypes 2, 3, 5, 6, and 8 in rutabaga (Brassica napus var. napobrassica)

Genome ◽

10.1139/gen-2016-0034 ◽

2016 ◽

Vol 59 (10) ◽

pp. 805-815 ◽

Cited By ~ 24

Author(s):

Muhammad Jakir Hasan ◽

Habibur Rahman

Keyword(s):

Brassica Napus ◽

Doubled Haploid ◽

Genetic Basis ◽

Molecular Mapping ◽

Plasmodiophora Brassicae ◽

Single Gene ◽

Genomic Region ◽

Clubroot Disease ◽

Dh Population ◽

Brassica Crops

Clubroot disease, caused by Plasmodiophora brassicae, is a threat to the production of Brassica crops including oilseed B. napus. In Canada, several pathotypes of this pathogen, such as pathotypes 2, 3, 5, 6, and 8, were identified, and resistance to these pathotypes was found in a rutabaga (B. napus var. napobrassica) genotype. In this paper, we report the genetic basis and molecular mapping of this resistance by use of F2, backcross (BC1), and doubled haploid (DH) populations generated from crossing of this rutabaga line to a susceptible spring B. napus canola line. The F1, F2, and BC1 populations were evaluated for resistance to pathotype 3, and the DH population was evaluated for resistance to pathotypes 2, 3, 5, 6, and 8. A 3:1 segregation in F2 and a 1:1 segregation in BC1 were found for resistance to pathotype 3, and a 1:1 segregation was found in the DH population for resistance to all pathotypes. Molecular mapping by using the DH population identified a genomic region on chromosome A8 carrying resistance to all five pathotypes. This suggests that a single gene or a cluster of genes, located in this genomic region, is involved in the control of resistance to these pathotypes.

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Soil microbiota influences clubroot disease by modulating Plasmodiophora brassicae and Brassica napus transcriptomes

Microbial Biotechnology ◽

10.1111/1751-7915.13634 ◽

2020 ◽

Vol 13 (5) ◽

pp. 1648-1672

Author(s):

Stéphanie Daval ◽

Kévin Gazengel ◽

Arnaud Belcour ◽

Juliette Linglin ◽

Anne‐Yvonne Guillerm‐Erckelboudt ◽

...

Keyword(s):

Brassica Napus ◽

Plasmodiophora Brassicae ◽

Clubroot Disease ◽

Soil Microbiota

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Identification of lncRNAs Responsive to Infection by Plasmodiophora brassicae in Clubroot-Susceptible and -Resistant Brassica napus Lines Carrying Resistance Introgressed from Rutabaga

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-12-18-0341-r ◽

2019 ◽

Vol 32 (10) ◽

pp. 1360-1377 ◽

Cited By ~ 3

Author(s):

Aarohi Summanwar ◽

Urmila Basu ◽

Habibur Rahman ◽

Nat Kav

Keyword(s):

Brassica Napus ◽

Target Genes ◽

Plasmodiophora Brassicae ◽

Genome Chromosome ◽

Clubroot Disease ◽

Brassica Crops ◽

A Genome ◽

Microbe Interactions ◽

Trait Locus ◽

Plant Pathogen Interactions

Clubroot disease, caused by Plasmodiophora brassicae Woronin, is a major threat to the production of Brassica’ crops. Resistance to different P. brassicae pathotypes has been reported in the A genome, chromosome A08; however, the molecular mechanism of this resistance, especially the involvement of long noncoding RNAs (lncRNAs), is not understood. We have used a strand-specific lncRNA-Seq approach to catalog lncRNAs from the roots of clubroot-susceptible and -resistant Brassica napus lines. In total, 530 differentially expressed (DE) lncRNAs were identified, including 88% of long intergenic RNAs and 11% natural antisense transcripts. Sixteen lncRNAs were identified as target mimics of the microRNAs (miRNAs) and eight were identified as the precursors of miRNAs. KEGG pathway analysis of the DE lncRNAs showed that the cis-regulated target genes mostly belong to the phenylpropanoid biosynthetic pathway (15%) and plant–pathogen interactions (15%) while the transregulated target genes mostly belong to carbon (18%) and amino acid biosynthesis pathway (19%). In all, 24 DE lncRNAs were identified from chromosome A08, which is known to harbor a quantitative trait locus conferring resistance to different P. brassicae pathotypes; however, eight of these lncRNAs showed expression only in the resistant plants. These results could form the basis for future studies aimed at delineating the roles of lncRNAs in plant–microbe interactions.

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Metabotyping: A New Approach to Investigate Rapeseed (Brassica napus L.) Genetic Diversity in the Metabolic Response to Clubroot Infection

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-02-12-0032-r ◽

2012 ◽

Vol 25 (11) ◽

pp. 1478-1491 ◽

Cited By ~ 15

Author(s):

Geoffrey Wagner ◽

Sophie Charton ◽

Christine Lariagon ◽

Anne Laperche ◽

Raphaël Lugan ◽

...

Keyword(s):

Amino Acids ◽

Brassica Napus ◽

Root System ◽

Metabolic Response ◽

Plasmodiophora Brassicae ◽

Late Response ◽

Primary Metabolism ◽

Brassica Napus L ◽

Clubroot Disease ◽

Metabolic Sink

Clubroot disease affects all Brassicaceae spp. and is caused by the obligate biotroph pathogen Plasmodiophora brassicae. The development of galls on the root system is associated with the establishment of a new carbon metabolic sink. Here, we aimed to deepen our knowledge of the involvement of primary metabolism in the Brassica napus response to clubroot infection. We studied the dynamics and the diversity of the metabolic responses to the infection. Root system metabotyping was carried out for 18 rapeseed genotypes displaying different degrees of symptom severity, under inoculated and noninoculated conditions at 42 days postinoculation (dpi). Clubroot susceptibility was positively correlated with clubroot-induced accumulation of several amino acids. Although glucose and fructose accumulated in some genotypes with minor symptoms, their levels were negatively correlated to the disease index across the whole set of genotypes. The dynamics of the metabolic response were studied for the susceptible genotype ‘Yudal,’ which allowed an “early” metabolic response (established from 14 to 28 dpi) to be differentiated from a “late” response (from 35 dpi). We discuss the early accumulation of amino acids in the context of the establishment of a nitrogen metabolic sink and the hypothetical biological role of the accumulation of glutathione and S-methylcysteine.

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Hormonal Responses to Plasmodiophora brassicae Infection in Brassica napus Cultivars Differing in Their Pathogen Resistance

International Journal of Molecular Sciences ◽

10.3390/ijms19124024 ◽

2018 ◽

Vol 19 (12) ◽

pp. 4024 ◽

Cited By ~ 10

Author(s):

Sylva Prerostova ◽

Petre Dobrev ◽

Veronika Konradyova ◽

Vojtech Knirsch ◽

Alena Gaudinova ◽

...

Keyword(s):

Brassica Napus ◽

Plasmodiophora Brassicae ◽

Marker Gene ◽

Pathogen Resistance ◽

Biosynthetic Gene ◽

Gall Formation ◽

Hormonal Responses ◽

Transient Elevation ◽

Indole 3 Acetic Acid ◽

Time Point

Hormonal dynamics after Plasmodiophora brassicae infection were compared in two Brassica napus cultivars—more resistant SY Alister and more sensitive Hornet, in order to elucidate responses associated with efficient defense. Both cultivars responded to infection by the early transient elevation of active cytokinins (predominantly cis-zeatin) and auxin indole-3-acetic acid (IAA) in leaves and roots, which was longer in Hornet. Moderate IAA levels in Hornet roots coincided with a high expression of biosynthetic gene nitrilase NIT1 (contrary to TAA1, YUC8, YUC9). Alister had a higher basal level of salicylic acid (SA), and it stimulated its production (via the expression of isochorismate synthase (ICS1)) in roots earlier than Hornet. Gall formation stimulated cytokinin, auxin, and SA levels—with a maximum 22 days after inoculation (dai). SA marker gene PR1 expression was the most profound at the time point where gall formation began, in leaves, roots, and especially in galls. Jasmonic acid (JA) was higher in Hornet than in Alister during the whole experiment. To investigate SA and JA function, SA was applied before infection, and twice (before infection and 15 dai), and JA at 15 dai. Double SA application diminished gall formation in Alister, and JA promoted gall formation in both cultivars. Activation of SA/JA pathways reflects the main differences in clubroot resistance.

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A New Identification Method Reveals the Resistance of an Extensive-Source Radish Collection to Plasmodiophora brassicae Race 4

Agronomy ◽

10.3390/agronomy11040792 ◽

2021 ◽

Vol 11 (4) ◽

pp. 792

Author(s):

Haohui Yang ◽

Yuxiang Yuan ◽

Xiaochun Wei ◽

Xiaohui Zhang ◽

Haiping Wang ◽

...

Keyword(s):

Plasmodiophora Brassicae ◽

Central China ◽

Clubroot Disease ◽

Cruciferous Vegetable ◽

Inoculation Methods ◽

Identification Method ◽

South Central ◽

Disease Occurrence ◽

Resistant Varieties ◽

The Stability

Raphanus sativus, an important cruciferous vegetable, has been increasingly affected by clubroot disease. Establishing a stable and accurate resistance identification method for screening resistant germplasms is urgently needed in radish. In this study, the influence of inoculum concentration, inoculation methods, and pH of the substrate on disease occurrence was studied. The result showed that the disease index (DI) was highest at 2 × 108 spores/mL, the efficiency of two-stage combined inoculation methods was higher than others, and pH 6.5 was favorable for the infection of P. brassicae. By using this new method, DIs of 349 radish germplasms varying from 0.00 to 97.04, presented significantly different levels of resistance. Analysis showed that 85.06% germplasms from China were susceptible to P. brassicae, whilst 28 accessions were resistant and mainly distributed in east, southwest, northwest, and south-central China. Most of the exotic germplasms were resistant. Repeated experiments verified the stability and reliability of the method and the identity of germplasm resistance. In total, 13 immune, 5 highly resistant and 21 resistant radish accessions were identified. This study provides an original clubroot-tolerance evaluation technology and valuable materials for the development of broad-spectrum resistant varieties for sustainable clubroot management in radish and other cruciferous crops.

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