Rapid detection of the pine wood nematode Bursaphelenchus xylophilus using recombinase polymerase amplification combined with CRISPR/Cas12a

Dai, Tingting; Chen, Zhenpeng; Guo, Yufang; Ye, Jian‐Ren

doi:10.1016/j.cropro.2023.106259

Cited by 2 publications

(2 citation statements)

References 50 publications

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“…A statistical analysis was performed using Graph Pad Prism 8 (Graph Pad Software Inc., San Diego, CA, USA). Student's t-test was used to compare the experimental and control groups, and only the differences with p-value < 0.05 were considered statistically significant [39].…”

Section: Rpa-crispr/cas12a Assaymentioning

confidence: 99%

Rapid Detection of Phytophthora cambivora Using Recombinase Polymerase Amplification Combined with CRISPR/Cas12a

Zhou,

Dai,

Dai

et al. 2023

Forests

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Phytophthora cambivora is a major quarantine pathogen that devastates economically important plants across the globe. P. cambivora causes ink disease in chestnut trees and root and stem rot in various fruit trees, resulting in significant yield reductions and plant death. Given the potential dangers of P. cambivora, effective detection methods are needed for both disease management and prevention. In this study, based on the whole-genome screening of specific target genes, a combination of the recombinase polymerase amplification technique (RPA) and CRISPR/Cas12 was established to detect P. cambivora. The RPA-CRISPR/Cas12a assay was able to specifically detect 7 target isolates of P. cambivora but did not detect the following 68 non-target isolates, including 28 isolates of Phytophthora, 3 isolates of Pythium, 3 isolates of Phytopythium, 32 isolates of fungi, and 2 isolates of Bursaphelenchus. The RPA-CRISPR/Cas12a detection method was able to detect 10 pg·μL−1 of P. cambivora genomic DNA at 37 °C within a short time span (60 min). Additionally, this method can identify the presence of P. cambivora in artificially inoculated apple fruits. In summary, compared with conventional detection techniques, the RPA-CRISPR/Cas12a detection method eliminates the need for expensive instruments, long reaction times, and high amounts of raw materials and can detect P. cambivora in imported plants at entry ports, enabling instant prevention and detection.

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Section: Rpa-crispr/cas12a Assaymentioning

confidence: 99%

Rapid Detection of Phytophthora cambivora Using Recombinase Polymerase Amplification Combined with CRISPR/Cas12a

Zhou,

Dai,

Dai

et al. 2023

Forests

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“…In addition, CRISPR technology is now being employed in forest tree species, such as poplar, 27 pine, 28 and spruce 29 to identify and develop gene-editing tools to modify the genetic makeup of trees. These tools have potential to improve the health and growth of trees by reducing their susceptibility to diseases, and altering the wood structure to create more valuable timber.…”

Section: Introduction/basic Gene Editing Strategymentioning

confidence: 99%

CRISPR/Cas9-gene editing approaches in plant breeding

Saini,

Thakur,

Gill

et al. 2023

GM Crops & Food

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CRISPR/Cas9 gene editing system is recently developed robust genome editing technology for accelerating plant breeding. Various modifications of this editing system have been established for adaptability in plant varieties as well as for its improved efficiency and portability. This review provides an in-depth look at the various strategies for synthesizing gRNAs for efficient delivery in plant cells, including chemical synthesis and in vitro transcription. It also covers traditional analytical tools and emerging developments in detection methods to analyze CRISPR/Cas9 mediated mutation in plant breeding. Additionally, the review outlines the various analytical tools which are used to detect and analyze CRISPR/Cas9 mediated mutations, such as next-generation sequencing, restriction enzyme analysis, and southern blotting. Finally, the review discusses emerging detection methods, including digital PCR and qPCR. Hence, CRISPR/Cas9 has great potential for transforming agriculture and opening avenues for new advancements in the system for gene editing in plants.

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Rapid detection of the pine wood nematode Bursaphelenchus xylophilus using recombinase polymerase amplification combined with CRISPR/Cas12a

Cited by 2 publications

References 50 publications

Rapid Detection of Phytophthora cambivora Using Recombinase Polymerase Amplification Combined with CRISPR/Cas12a

Rapid Detection of Phytophthora cambivora Using Recombinase Polymerase Amplification Combined with CRISPR/Cas12a

CRISPR/Cas9-gene editing approaches in plant breeding

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