A rapid approach for isolating a single fungal spore from rice blast diseased leaves

Fei, Li-wang; Lu, Wenbo; Xu, Xiao-zhou; Feng-cheng, Yan; Zhang, Liwei; Liu, Jintao; Bai, Yuanjun; Li, Zhenyu; Zhao, Wensheng; Yang, Jun; Peng, You-Liang

doi:10.1016/s2095-3119(19)62581-5

Cited by 16 publications

(10 citation statements)

References 12 publications

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“…Field samples are always contaminated with various fastgrowing necrotrophic fungi, which makes the isolation of MoT very complicated. Moreover, fetching infected field samples to the lab requires long distance travel, thus increasing the chance of contamination (Fei et al 2019). Despite that the appropriate surface sterilization measure has been taken, various mold fungi, Alternaria spp., Fusarium spp., Curvularia spp.…”

Section: Discussionmentioning

confidence: 99%

Suitable methods for isolation, culture, storage and identification of wheat blast fungus Magnaporthe oryzae Triticum pathotype

et al. 2020

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Wheat blast disease caused by a South American lineage of Magnaporthe oryzae Triticum (MoT) pathotype has emerged as a serious threat to wheat production in Bangladesh since its first emergence in 2016. Efficient and suitable methods for isolation, storage, inoculum production and molecular characterization of the pathogen can help in achieving the target of sustainable management of the disease in a relatively short period of time. In this study, we aimed to develop suitable methods for isolation, storage and morphological characterization and molecular identification of MoT isolates collected from the blast-infected wheat fields in Bangladesh. This process included modification of existing protocols that were available for a related fungal pathogen M. oryzae or de novo method development and validation. We developed suitable methods for isolation of MoT from field-infected plant samples using modified monoconidial isolation technique and produced abundant conidia from a single mycelial plate for in vivo pathogenicity assay in a reproducible manner. Cultural and morphological characterization of the isolates revealed that all Bangladeshi MoT isolates are of a single clonal lineage with similar cultural and morphological characters. Molecular detection of isolates with M. oryzae-specific primers Pot1 and Pot2 and MoT-specific primers MoT3F and MoT3R produced bands with the expected size from all wheat-infecting isolates. We also successfully established a PCR-based detection system based on a commercially available detection kit for field-infected leaf and seed samples by detecting Pot2- and MoT3-specific bands. Additionally, the simple method we developed in our study for producing abundant conidia in a very short period of time will be very helpful in studying biology of the wheat blast fungus. This method was also proven to be more user-friendly and cost-effective than previously available methods. Successful characterization of MoT isolates at morphological and molecular levels coupled with detection of the pathogen in infected field and seed lots should be useful for efficient surveillance and management of the fearsome wheat blast disease.

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Section: Discussionmentioning

confidence: 99%

Suitable methods for isolation, culture, storage and identification of wheat blast fungus Magnaporthe oryzae Triticum pathotype

et al. 2020

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“…Although the individual sizes were not exactly the same, they did not differ too much, and the size and shape of a single individual hardly changed. 29 Therefore, the diffractive fingerprint of the detected microparticles is only the diffractive fingerprint of rice blast spores, whose size and texture are similar.…”

Section: Results Of the Device Evaluationmentioning

confidence: 99%

A rapid rice blast detection and identification method based on crop disease spores' diffraction fingerprint texture

Yang

Wang

et al. 2020

J Sci Food Agric

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BACKGROUND Rice blast fungus is a worldwide disease, and it is one of the most serious rice diseases in the north and south rice fields in China. The initial symptoms of rice blast are not obvious, and the speed of transmission is fast. Manual identification is time‐consuming and laborious. At present, it is a great challenge to realize rapid and accurate early identification of rice blast. RESULTS In this paper, an identification method based on crop disease spores' diffraction fingerprint texture for rice blast was studied; this method utilizes the light field and texture features of diffraction images. To verify the reliability of the model that we proposed, we selected two methods of manual identification and machine recognition to compare and detect rice blast spores. The experimental results show that the identification of light diffraction characteristics is not only higher than the traditional manual recognition by microscope (increased by more than 0.3%), but also faster after neural network training (increased by more than 90%). The diffraction recognition method used in this study, based on crop disease spores' diffraction fingerprint texture, can be completed in a few seconds, and its test accuracy is 97.18%. CONCLUSION The proposed method, a rapid rice blast detection and identification method based on crop disease spores' diffraction fingerprint texture, has certain advantages compared with the existing manual identification by microscope. This method can be applied to the recognition of rice blast in agricultural research. © 2020 Society of Chemical Industry

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“…There are many methods to cultivate the conidia of Magnaporthe grisea (Fei et al, 2019; Zhang, Su, & Cai, 2013). In the laboratory, two methods are usually used.…”

Section: Methodsmentioning

confidence: 99%

A rapid detection method of early spore viability based on AC impedance measurement

Yang

et al. 2020

J Food Process Engineering

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Rice blast is a destructive disease of rice; it is difficult to detect the pathogenic spore viability in early stage. In this paper, a rapid detection method is proposed; the impedance‐viability model of spores was established by measuring the viability of early rice blast spores and the AC impedance at the most sensitive frequencies. This model is a nice linear model, and the goodness of fit R2 is greater than 0.99. At the same time, there is a positive correlation between the spore impedance‐viability model and the hyphae impedance‐growth speed model measured in practice. The p value is less than 0.05, and the Pearson coefficient is close to 1, which proves the feasibility of this method to detect spore viability. Compared with the traditional experimental culture method, the detection speed of spore viability based on impedance method is greatly improved, which brings convenience for the study of rice blast. Practical Applications The viability of spores is one of the most important indexes to show the degree of rice blast. The early spores of the rice blast look similarly, which makes it hard for the naked eye to distinguish the viability, and makes it difficult for farmers to decide whether to use pesticides in the rice field. The traditional method for the detection of spore viability of rice blast is complicated and needs a long time. In recent years, the method of microbial impedance is put forward, which can provide the information of microbial AC impedance, and has the advantages of fast and nondestructive. The results showed that it is feasible to detect spore viability by impedance technique.

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A rapid approach for isolating a single fungal spore from rice blast diseased leaves

Cited by 16 publications

References 12 publications

Suitable methods for isolation, culture, storage and identification of wheat blast fungus Magnaporthe oryzae Triticum pathotype

Suitable methods for isolation, culture, storage and identification of wheat blast fungus Magnaporthe oryzae Triticum pathotype

A rapid rice blast detection and identification method based on crop disease spores' diffraction fingerprint texture

A rapid detection method of early spore viability based on AC impedance measurement

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