scholarly journals Plasmodiophora brassicae infection threshold – How many resting spores are required for infection of canola (Brassica napus)

2021 ◽  
Author(s):  
Kher Zahr ◽  
Yalong Yang ◽  
Snezana Dijanovic ◽  
Heting Fu ◽  
Michael Harding ◽  
...  
Keyword(s):  
Plasmodiophora Brassicae ◽  
Management Strategies ◽  
Detection Methods ◽  
Single Spore ◽  
Clubroot Disease ◽  
Highly Sensitive ◽  
Resting Spores ◽  
Low Probability ◽  
Highly Sensitive Detection ◽  
Important Disease

Clubroot, caused by Plasmodiophora brassicae, is an important disease of canola and other brassica crops. Improved understanding of host and pathogen biology is frequently useful in guiding management strategies. In order to better understand infection thresholds, seven-day old seedlings of canola cultivar Westar were inoculated with serially diluted concentrations of P. brassicae resting spores. Controlled soil and plant inoculation assays were performed and the plants maintained in a greenhouse for 42 days and clubroot disease severity evaluated visually. Clubroot symptoms were observed in soils containing as low as one spore/mL soil and on plants inoculated with as few as ≤ 100 resting spores. These thresholds were lower than any previously reported. The results indicated the importance of highly sensitive detection methods for P. brassicae diagnosis and quantification methods for clubroot risk prediction in soils. Furthermore, these results highlighted the low probability of obtaining P. brassicae single spore isolates.

scholarly journals Isolation and Variation in Virulence of Single-Spore Isolates of Plasmodiophora brassicae from Canada

Plant Disease ◽  
2008 ◽  
Vol 92 (3) ◽  
pp. 456-462 ◽  
Author(s):  
S. Xue ◽  
T. Cao ◽  
R. J. Howard ◽  
S. F. Hwang ◽  
S. E. Strelkov
Keyword(s):  
British Columbia ◽  
Brassica Napus ◽  
Efficient Method ◽  
Plasmodiophora Brassicae ◽  
Breeding Strategy ◽  
Single Spore ◽  
Susceptible Host ◽  
Significant Difference ◽  
Resting Spores ◽  
Important Disease

Clubroot of crucifers, caused by Plasmodiophora brassicae, is emerging as an important disease of canola (Brassica napus) in Alberta, Canada. Populations of the pathogen often consist of a mixture of different pathotypes. Therefore, a simple and efficient method to isolate single resting spores of P. brassicae was developed, based on serial dilution of spore suspensions. The virulence of 24 single-spore isolates, representing five populations of the pathogen from Alberta, Ontario, and British Columbia, was characterized on the differentials of Williams and Somé et al. Symptoms were rated 6 weeks after inoculation and Fisher's least significant difference (P < 0.05) was used to differentiate resistant from susceptible host reactions. The pathotype composition of P. brassicae in Canada appeared more diverse when single-spore isolates were examined rather than populations of the pathogen. In Alberta, at least three and possibly four pathotypes were identified among the 14 isolates tested, whereas a maximum of only two pathotypes had been reported previously when populations of the pathogen were examined. Pathotype 3 or P2, as classified on the differentials of Williams and Somé et al., respectively, was found to be predominant in the province. The occurrence of other pathotypes at lower frequencies suggests that caution should be used in any breeding strategy, because rare pathotypes of P. brassicae may quickly become predominant if susceptible host genotypes are continuously grown.

scholarly journals Effect of pathogen virulence on pathogenicity, host range and reproduction of Plasmodiophora brassicae, the causal agent of clubroot disease

Plant Disease ◽  
2021 ◽  
Author(s):  
Nazanin Zamani-Noor ◽  
Sinja Brand ◽  
Hans-Peter Soechting
Keyword(s):  
Plasmodiophora Brassicae ◽  
Brassica Species ◽  
Post Inoculation ◽  
Clubroot Disease ◽  
Alopecurus Myosuroides ◽  
Pathogen Virulence ◽  
Iberis Amara ◽  
Thlaspi Arvense ◽  
Resting Spores ◽  
Pathogen Dna

A series of greenhouse experiments was conducted to evaluate the effect of Plasmodiophora brassicae virulence on clubroot development and propagation of resting spores in 86 plant species from 19 botanical families. Plants were artificially inoculated with two isolates of P. brassicae, which were either virulent on clubroot-resistant oilseed rape cv. Mendel (P1 (+)) or avirulent on this cultivar (P1). Clubroot severity and the number of resting spores inside the roots were assessed 35 days post inoculation. Typical clubroot symptoms were observed only in the Brassicaceae family. P1 (+)-inoculated species exhibited more severe symptoms (2 to 10–fold more severe), bigger galls (1.1 to 5.8 fold heavier) and higher number of resting spores than the P1-inoculated plants. Among all Brassica species, Bunias orientalis, Coronopus squamatus and Raphanus sativus were fully resistant against both isolates, while Camelina sativa, Capsella bursa-pastoris, Coincya momensis, Descurainia sophia, Diplotaxis muralis, Erucastrum gallicum, Neslia paniculata, Sinapis alba, S. arvensis, Sisymbrium altissimum, S. loeselii and Thlaspi arvense were highly susceptible. Conringia orientalis, Diplotaxis tenuifolia, Hirschfeldia incana, Iberis amara, Lepidium campestre and Neslia paniculata were completely or partially resistant to P1-isolate but highly susceptible to P1 (+). These results propose that the basis for resistance in these species may be similar to that found in some commercial cultivars, and that these species could contribute to the build-up of inoculum of virulent pathotypes. Furthermore, the pathogen DNA was detected in Alopecurus myosuroides, Phacelia tanacatifolia, Papaver rhoeas and Pisum sativum. It can concluded that the number and diversity of hosts for P. brassicae are greater than previously reported.

A novel NIR fluorescent probe for highly sensitive detection of HNO and its application in bioimaging

2021 ◽  
Author(s):  
shenwei he ◽  
Jianming Zhu ◽  
peiyao xie ◽  
Jianfei Liu ◽  
Di Zhang ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Reactive Nitrogen Species ◽  
Physiological Process ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Reactive Nitrogen ◽  
Nitrogen Species ◽  
Efficient Detection ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Nitroxyl (HNO) as an important reactive nitrogen species (RNS), which plays an important role in multiple physiological process. Therefore, it is urgent to exploit reliable and efficient detection methods for...

scholarly journals Virulence Spectrum of Single-Spore and Field Isolates of Plasmodiophora brassicae Able to Overcome Resistance in Canola (Brassica napus)

Plant Disease ◽  
2021 ◽  
Vol 105 (1) ◽  
pp. 43-52 ◽  
Author(s):  
Homa Askarian ◽  
Alireza Akhavan ◽  
Victor P. Manolii ◽  
Tiesen Cao ◽  
Sheau-Fang Hwang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Confidence Interval ◽  
Plasmodiophora Brassicae ◽  
Large Collection ◽  
Single Spore ◽  
Brassica Napus L ◽  
Clubroot Resistance ◽  
Field Isolates ◽  
Virulence Spectrum ◽  
Important Disease

Clubroot, caused by Plasmodiophora brassicae Woronin, is an important disease of canola (Brassica napus L.) that is managed mainly by planting clubroot-resistant (CR) cultivars. Field isolates of P. brassicae can be heterogeneous mixtures of various pathotypes, making assessments of the genetics of host–pathogen interactions challenging. Thirty-four single-spore isolates were obtained from nine field isolates of the pathogen collected from CR canola cultivars. The virulence patterns of the single-spore and field isolates were assessed on the 13 host genotypes of the Canadian Clubroot Differential (CCD) set, which includes the differentials of Williams and Somé et al. Indices of disease (IDs) severity of 25, 33, and 50% (±95% confidence interval) were compared as potential thresholds to distinguish between resistant and susceptible reactions, with an ID of 50% giving the most consistent responses for pathotype classification purposes. With this threshold, 13 pathotypes could be distinguished based on the CCD system, 7 on the differentials of Williams, and 3 on the hosts of Somé et al. The highest correlations were observed among virulence matrices generated using the three threshold IDs on the CCD set. Genetically homogeneous single-spore isolates gave a clearer profile of the P. brassicae pathotype structure. Novel pathotypes, not reported in Canada previously, were identified among the isolates. This large collection of single-spore isolates can serve as a reference in screening and breeding for clubroot resistance.

scholarly journals Characterization of Five Molecular Markers for Pathotype Identification of the Clubroot Pathogen Plasmodiophora brassicae

2018 ◽  
Vol 108 (12) ◽  
pp. 1486-1492 ◽  
Author(s):  
Jing Zheng ◽  
Xuliang Wang ◽  
Qian Li ◽  
Shu Yuan ◽  
Shiqing Wei ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Plasmodiophora Brassicae ◽  
Expression Patterns ◽  
Chain Reaction ◽  
Clubroot Disease ◽  
Polymerase Chain ◽  
Differential Hosts ◽  
Reaction Cycles ◽  
Important Disease

Clubroot disease is an important disease on cruciferous crops caused by Plasmodiophora brassicae infections. The pathotypes have been classified based on the reactions of differential hosts. However, molecular markers of particular pathotypes for P. brassicae are limited. In this study, we found five genetic markers in association with different pathotypes. Different gene expression patterns among different pathotypes (P4, P7, P9, and P11) were assayed according to the transcriptome data. The assay indicated that molecular markers PBRA_007750 and PBRA_009348 could be used to distinguish P11 from P4, P7, and P9; PBRA_009348 and Novel342 could distinguish P9 from P4, P7, and P11; and PBRA_008439 and Novel342 could represent a kind of P4. Polymerase chain reaction cycles ranging from 25 to 30 were able to identify the predominant pathotype in general. Therefore, these molecular markers would be a valuable tool to identify and discriminate pathotypes in P. brassicae population.

scholarly journals First Report of Clubroot on Capsella bursa-pastoris Caused by Plasmodiophora brassicae in Sichuan Province of China

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 687-687 ◽  
Author(s):  
L. Ren ◽  
X. P. Fang ◽  
C. C. Sun ◽  
K. R. Chen ◽  
F. Liu ◽  
...  
Keyword(s):  
Plasmodiophora Brassicae ◽  
Tap Water ◽  
Disease Incidence ◽  
Morphological Characteristics ◽  
Pcr Amplification ◽  
Sichuan Province ◽  
Clubroot Disease ◽  
Cruciferous Vegetable ◽  
First Report ◽  
Resting Spores

Shepherd's purse (Capsella bursa-pastoris (L.) Medicus) is an edible and wild medicinal plant widely distributed in China. This plant has been cultivated in Shanghai, China, since the end of the 19th century. Infection of C. bursa-pastoris by Plasmodiophora brassicae, the causal agent of clubroot disease on Brassica spp. has been reported in Korea (2), but is not known to occur in China. In February of 2011, stunted and wilted shepherd's purse (SP) plants were observed in a field planted to oilseed rapes (B. napus) in Sichuan Province of China. Symptomatic SP plants also exhibited root galls. Disease incidence was 6.2% and 100% for SP and B. napus, respectively. Root galls on diseased SP plants were collected for pathogen identification. Many resting spores were observed when the root galls were examined under a light microscope. The resting spores were circular in shape, measuring 2.0 to 3.1 μm in diameter (average 2.6 μm). PCR amplification was conducted to confirm the pathogen. DNA was extracted from root galls and healthy roots (control) of SP. Two primers, TC2F (5′-AAACAACGAGTCAGCTTGAATGCTAGTGTG-3′) and TC2R (5′-CTTTAGTTGTGTTTCGGCTAGGATGGTTCG-3′) were used to detect P. brassicae (1). No PCR amplifications were observed with the control DNA as template. A fragment of the expected size (approximately 520 bp) was obtained when DNA was amplified from diseased roots of SP. These results suggest that the pathogen in the galled roots of SP is P. brassicae. Pathogenicity of P. brassicae in SP was tested on plants of both SP and Chinese cabbage (CC) (B. campestris ssp. pekinensis). A resting spore suspension prepared from naturally infected SP roots was mixed with a sterilized soil in two plastic pots, resulting in a final concentration of 5 × 106 spores/g soil. Soil treated with the same volume of sterile water was used as a control. Seeds of SP and CC were pre-germinated on moist filter paper for 2 days (20°C) and seeded into the infested and control pots, one seed per pot for planted for CC and four seeds per pot for SP. The pots were placed in a chamber at 15 to 25°C under 12 h light and 12 h dark. Plants in each pot were uprooted after 4 weeks and the roots of each plant were washed under tap water and rated for clubroot disease. No disease symptoms were observed in the control treatments of SP or CC. Plants of both species showed symptoms of clubroot, with the disease incidence of 62.5% and 100% on SP and CC, respectively. The pathogen was isolated from diseased roots of each plant and confirmed as P. brassicae based on morphological characteristics and PCR detection. To our knowledge, this is the first report of clubroot disease on C. bursa-pastoris in Sichuan Province of China. This finding suggests that it may be necessary to manage C. bursa-pastoris in cruciferous vegetable (cabbage, turnip) and oilseed rape production fields. References: (1) T. Cao et al. Plant Dis. 91:80, 2007. (2) W. G. Kim et al. Microbiology 39:233, 2011.

scholarly journals Biosensors for Deoxynivalenol and Zearalenone Determination in Feed Quality Control

Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 499
Author(s):  
Krisztina Majer-Baranyi ◽  
Nóra Adányi ◽  
András Székács
Keyword(s):  
Sensitive Detection ◽  
Farm Animals ◽  
Detection Methods ◽  
Sample Treatment ◽  
Minimal Sample ◽  
Current Trends ◽  
Highly Sensitive ◽  
Feed Analysis ◽  
Highly Sensitive Detection ◽  
Analytical Tools

Mycotoxin contamination of cereals used for feed can cause intoxication, especially in farm animals; therefore, efficient analytical tools for the qualitative and quantitative analysis of toxic fungal metabolites in feed are required. Current trends in food/feed analysis are focusing on the application of biosensor technologies that offer fast and highly selective and sensitive detection with minimal sample treatment and reagents required. The article presents an overview of the recent progress of the development of biosensors for deoxynivalenol and zearalenone determination in cereals and feed. Novel biosensitive materials and highly sensitive detection methods applied for the sensors and the application of these sensors to food/feed products, the limit, and the time of detection are discussed.

scholarly journals Nanozyme-Participated Biosensing of Pesticides and Cholinesterases: A Critical Review

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 382
Author(s):  
Hengjia Zhu ◽  
Peng Liu ◽  
Lizhang Xu ◽  
Xin Li ◽  
Panwang Hu ◽  
...  
Keyword(s):  
Environmental Remediation ◽  
Low Cost ◽  
Catalytic Performance ◽  
Agricultural Products ◽  
Detection Methods ◽  
Highly Sensitive ◽  
Potential Applications ◽  
Highly Sensitive Detection ◽  
Excellent Stability

To improve the output and quality of agricultural products, pesticides are globally utilized as an efficient tool to protect crops from insects. However, given that most pesticides used are difficult to decompose, they inevitably remain in agricultural products and are further enriched into food chains and ecosystems, posing great threats to human health and the environment. Thus, developing efficient methods and tools to monitor pesticide residues and related biomarkers (acetylcholinesterase and butylcholinesterase) became quite significant. With the advantages of excellent stability, tailorable catalytic performance, low cost, and easy mass production, nanomaterials with enzyme-like properties (nanozymes) are extensively utilized in fields ranging from biomedicine to environmental remediation. Especially, with the catalytic nature to offer amplified signals for highly sensitive detection, nanozymes were finding potential applications in the sensing of various analytes, including pesticides and their biomarkers. To highlight the progress in this field, here the sensing principles of pesticides and cholinesterases based on nanozyme catalysis are definitively summarized, and emerging detection methods and technologies with the participation of nanozymes are critically discussed. Importantly, typical examples are introduced to reveal the promising use of nanozymes. Also, some challenges in the field and future trends are proposed, with the hope of inspiring more efforts to advance nanozyme-involved sensors for pesticides and cholinesterases.

scholarly journals Methods for Assessment of Viability and Germination of Plasmodiophora brassicae Resting Spores

2022 ◽  
Vol 12 ◽  
Author(s):  
Yao Wang ◽  
Birger Koopmann ◽  
Andreas von Tiedemann
Keyword(s):  
Evans Blue ◽  
Plasmodiophora Brassicae ◽  
Germination Rate ◽  
Inoculum Potential ◽  
Differential Interference Contrast Microscopy ◽  
Calcofluor White ◽  
Clubroot Disease ◽  
Resting Spores ◽  
Dual Staining

Clubroot caused by the obligate biotrophic parasite Plasmodiophora brassicae is a destructive soil borne disease of cruciferous crops. Resting spores of P. brassicae can survive in the soil for a long period without hosts or external stimulants. The viability and germination rate of resting spores are crucial factors of the inoculum potential in the field. The accurate assessment of viability and germination rate is the foundation to evaluate the effect of control methods. In this study, we evaluated several methods for the assessment of viability and germination rate of P. brassicae resting spores. Dual staining with calcofluor white-propidium iodide (CFW-PI) or single stain with Evans blue showed reliable accuracy in estimating viability. CFW-PI was capable of reliably determining the viability within 10 min, while Evans blue required overnight incubation to obtain accurate results. Due to DNA degradation of heat treatments, acetone was selected to evaluate the efficiency of propidium monoazide (PMA)–quantitative PCR (qPCR) used for the quantification of DNA from viable cells. The staining with 4,6-Diamidine-2-phenylindole dihydrochloride (DAPI) and the use of differential interference contrast microscopy were suitable for the determination of resting spore germination rates. The latter method also allowed recording individual germination states of spores. Alternatively, dual staining with CFW-Nile red was successfully used to assess the germination rate of resting spores with a lethal pre-treatment. This study evaluates and confirms the suitability of various microscopic and molecular genetic methods for the determination of viability and germination of P. brassicae resting spores. Such methods are required to study factors in the soil regulating survival, dormancy and germination of P. brassicae resting spores causing clubroot disease in Brassicaceae hosts and therefore are fundamental to develop novel strategies of control.

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