Sensitive and high-throughput polyclonal antibody-based serological methods for rice stripe virus detection in both rice and small brown planthopper

Zhang, Kun; Zhuang, Xinjian; Xu, Hongmei; Guo, Xiao; He, Zhen; Xu, Kai; Liu, Fang

doi:10.1016/j.cropro.2021.105599

Cited by 7 publications

(11 citation statements)

References 13 publications

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“…The positive clone was selected using PCR, and His-tagged FvFUM21 and FvFUM21 175–521 were expressed through 1 mM isopropyl ß-D-1-thiogalactopyranoside (IPTG) induction, and purified with the Ni-NTA 6FF Sefinose Resin Kit (Cat. no., C600332, Sangon Biotech, Shanghai, China), as described previously with minor modifications …”

Section: Methodsmentioning

confidence: 99%

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Discovery and Development of Luvangetin from Zanthoxylum avicennae as a New Fungicide Candidate for Fusarium verticillioides

Chen,

Zhou,

Liu

et al. 2024

J. Agric. Food Chem.

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Pathogenic fungi pose a significant threat to crop yields and human healthy, and the subsequent fungicide resistance has greatly aggravated these agricultural and medical challenges. Hence, the development of new fungicides with higher efficiency and greater environmental friendliness is urgently required. In this study, luvangetin, isolated and identified from the root of Zanthoxylum avicennae, exhibited wide-spectrum antifungal activity in vivo and in vitro. Integrated omics and in vitro and in vivo transcriptional analyses revealed that luvangetin inhibited GAL4-like Zn(II) 2 Cys6 transcriptional factor-mediated transcription, particularly the FvFUM21-mediated FUM cluster gene expression, and decreased the biosynthesis of fumonisins inFusarium verticillioides. Moreover, luvangetin binds to the double-stranded DNA helix in vitro in the groove mode. We isolated and identified luvangetin, a natural metabolite from a traditional Chinese edible medicinal plant and uncovered its multipathogen resistance mechanism. This study is the first to reveal the mechanism underlying the antifungal activity of luvangetin and provides a promising direction for the future use of plant-derived natural products to prevent and control plant and animal pathogenic fungi.

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Section: Methodsmentioning

confidence: 99%

“…no., C600332, Sangon Biotech, Shanghai, China), as described previously with minor modifications. 18 Electrophoresis Mobility Shift Assay. A 455-bp-length DNA sequence was amplified from the predicted promoter of FvFUM13 transcript and treated for probe (FvFUM13 Pro -probe) by 5′biotinylation labeling using PCR (Tables S7 and S8).…”

Section: T Cmentioning

confidence: 99%

Discovery and Development of Luvangetin from Zanthoxylum avicennae as a New Fungicide Candidate for Fusarium verticillioides

Chen,

Zhou,

Liu

et al. 2024

J. Agric. Food Chem.

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“…The construction method of the prokaryotic expression vector was as follows, referring to the previous method [ 18 ]. Total RNA was extracted from RBSDV-infected rice seedlings and cDNA was generated using Takara PrimeScript RT Reagent Kit reverse transcription (Takara, Dalian, China).…”

Section: Methodsmentioning

confidence: 99%

Development of Polyclonal Antibodies and a Serological-Based Reverse-Transcription Loop-Mediated Isothermal Amplification (S-RT-LAMP) Assay for Rice Black-Streaked Dwarf Virus Detection in Both Rice and Small Brown Planthopper

Hua,

Feng,

et al. 2023

Viruses

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Rice black-streaked dwarf virus (RBSDV) infects rice and maize, and seriously affects rice yields in main rice-producing areas. It can be transmitted via small brown planthopper (SBPH: Laodelphax striatellus Fallén). To more rapidly, sensitively, and highly throughput diagnose RBSDV in the wild condition, we first purified the recombinant His-CPRBSDV protein, and prepared the polyclonal antibodies against the His-CPRBSDV protein (PAb-CPRBSDV). Based on the PAb-CPRBSDV, we developed a series of serological detections, such as Western blot, an enzyme-linked immunosorbent assay (ELISA), and a dot immunoblotting assay (DIBA). Furthermore, we developed a serological-based reverse-transcription loop-mediated isothermal amplification assay (S-RT-LAMP) that could accurately detect RBSDV in the wild. Briefly, the viral genomic dsRNA together with viral CP were precipitated by co-immunoprecipitation using the PAb-CPRBSDV, then the binding RNAs were crudely isolated and used for RT-LAMP diagnosis. Using the prepared PAb-CPRBSDV, four serology-based detection methods were established to specifically detect RBSDV-infected rice plants or SBPHs in the wild. The method of S-RT-LAMP has also been developed to specifically, high-throughput, and likely detect RBSDV in rice seedlings and SBPHs simultaneously. The antiserum prepared here laid the foundation for the rapid and efficient detection of RBSDV-infected field samples, which will benefit for determination of the virulence rate of the transmission vector SBPH and outbreak and epidemic prediction of RBSDV in a rice production area.

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“…Following immunization, the 1 mL rabbit blood was drawn from the ear vein, and the antiserum was placed at 4 • C overnight for titer evaluation. Total crude antisera against CP ReMV were obtained after centrifugation at 12,000 × g for 15 min, as previously described (Zhang et al, 2021). Sodium sulfate solution (50, 33, and 30%) was used to precipitate immunoglobulin IgG.…”

Section: Preparation Of Polyclonal Antibodies Against the His-cp Remv...mentioning

confidence: 99%

“…The PAb-CP ReMV was used to establish the western blotting, immuno-dot blotting, and ELISA methods for ReMV detection form R. glutinosa. All performances followed standard protocols (Barker, 1998;Zhang et al, 2021).…”

Section: Western Blotting Immuno-dot Blotting and Elisa Detection Of ...mentioning

confidence: 99%

Development of polyclonal antibodies-based serological methods for detection of the rehmannia mosaic virus in field plants

Chen

Feng

Guo

et al. 2022

Front. Sustain. Food Syst.

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Rehmannia glutinosa is a top-grade traditional Chinese medicine, and also is an important planting medicinal material for Chinese poor farmers shaking off poverty. Rehmannia mosaic virus (ReMV) causes big economic loss of R. glutinosa in planting area. However, there is no effective methods for quick, accurate, and high-throughput detection for ReMV in Chinese production area. The preserved R. glutinosa samples carrying ReMV was taken for research material. The coat protein coding sequences (CPReMV) was cloned and sequenced. The target sequence was further placed into a prokaryotic expression vector to express the N-terminal-tagged recombinant CPReMV protein (His-CPReMV). Purified His-CPReMV was used as an antigen to immunize New Zealand white rabbits, and antiserum was obtained. The titers and sensitivities of the antisera were analyzed and evaluated. Polyclonal antibodies were purified from the antiserum, and the titers and sensitivity to the target His-CPReMV protein were evaluated. The results demonstrate that the obtained polyclonal antibodies against His-CPReMV could be successfully used for rapid, accurate, and high-throughput detection of ReMV from R. glutinosa planted in the wild. Our investigation established serological-based detection methods for ReMV for the first time, and provides a foundation for future exploration of the pathogenic mechanisms of ReMV in R. glutinosa.

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Sensitive and high-throughput polyclonal antibody-based serological methods for rice stripe virus detection in both rice and small brown planthopper

Cited by 7 publications

References 13 publications

Discovery and Development of Luvangetin from Zanthoxylum avicennae as a New Fungicide Candidate for Fusarium verticillioides

Discovery and Development of Luvangetin from Zanthoxylum avicennae as a New Fungicide Candidate for Fusarium verticillioides

Development of Polyclonal Antibodies and a Serological-Based Reverse-Transcription Loop-Mediated Isothermal Amplification (S-RT-LAMP) Assay for Rice Black-Streaked Dwarf Virus Detection in Both Rice and Small Brown Planthopper

Development of polyclonal antibodies-based serological methods for detection of the rehmannia mosaic virus in field plants

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