A method for DNA extraction and molecular identification of Aphids

MethodsX, Journal Year: 2023, Volume and Issue: 10, P. 102100 - 102100

Published: Jan. 1, 2023

Aphid species (Insecta, Hemiptera) are economically important invasive pest throughout the world, though their identification is intricate due to tiny size and inconspicuous nature of morphology. Mitochondrial cytochrome c oxidase I (mtCOI) region has been proven be a standard barcode identify diverse array insect groups. Isolation good quality DNA fundamental first step in barcoding which obtained by standardizing isolation method. In this study, we demonstrate modified CTAB method for maximize yield from small aphids. This will help researchers efficiently isolate aphid can utilized other insects as well. We evaluated isolated mtCOI gene were subjected PCR amplification. Further, segment was sequenced annotation done NCBI BLAST program through found Aphis gossypii. study provides set molecular tools that used at level biodiversity analysis.•Detailed quantity genomic aphids.•Molecular aphids using amplification sequence validation.•First report on gossypii infecting Solanum trilobatum insights management.

Language: Английский

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Diversity in thrips palmi: Exploration of genetic and ecological heterogeneity

Gene, Journal Year: 2025, Volume and Issue: unknown, P. 149252 - 149252

Published: Jan. 1, 2025

Language: Английский

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Simple DNA extraction for museum beetle specimens to unlock genetic data from historical collections

MethodsX, Journal Year: 2025, Volume and Issue: 14, P. 103236 - 103236

Published: Feb. 28, 2025

Language: Английский

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On‐site genetic diagnosis for the invasive pest Hylurgus ligniperda (Fabricius) and its possible application

Pest Management Science, Journal Year: 2025, Volume and Issue: unknown

Published: March 13, 2025

Abstract BACKGROUND Forests in nearly all regions worldwide are affected by invasions of non‐native bark beetles. Hylurgus ligniperda (Fabricius) is a globally invasive beetle that stealthily jeopardizes pine health and spreads insidiously. The occurrence challenges trade logs or wooden materials. Early identification crucial implementing appropriate pest management strategies. RESULTS This study established simple, efficient, accurate method for identifying based on recombinase polymerase amplification the lateral flow dipstick ( RPA ‐ LFD ). can distinguish from other species has sensitivity threshold 10 fg/ μL . Subsequently, field application tests were conducted using assays, first case forestry entomology. test results showed combined with crude DNA extraction could accurately identify (except elytra). influence environmental factors (temperature, humidity, wind) was also investigated. only wind speed P = 0.003) significantly correlated color rendering negatively density detection line. CONCLUSION A rapid field‐based applied helps elimination barriers to lagging pests. Implementing RPA‐LFD aims provide reliable efficient tool rapidly insects, enabling timely intervention effective © 2025 Society Chemical Industry.

Language: Английский

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Low-cost water boiling method successfully employed to extract DNA from a single whitefly to detect yellow mosaic disease-causing viruses in PCR

Archives of Phytopathology and Plant Protection, Journal Year: 2024, Volume and Issue: 57(7), P. 509 - 525

Published: April 20, 2024

A cost-effective DNA extraction approach was developed and applied to PCR-based identification of yellow mosaic disease (YMD) causing viruses in individual whiteflies. Whiteflies were collected from urdbean eggplant, representing host non-host plants target viruses, respectively. whiteflies successfully extracted using the nuclease-free water boiling method optimised template concentration for PCR assays. An average nucleic acid content 3.844 ± 0.14 ng/µl 50 µl water. assays revealed Mungbean India virus (MYMIV) presence 90% urdbean. Conversely, YMD-causing not present eggplant or themselves. The use this low-cost detecting is very effective surveillance monitoring viruses. Early detection facilitates deployment appropriate management strategies, reducing both crop losses potential outbreaks.

Language: Английский

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FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols

Published: Jan. 18, 2024

Versatility, sensitivity and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research as well diagnostic applications. However, point-of-care (PoC) use traditional qPCR faces two main challenges: long run times mean results are not available half an hour or more requisite high-temperature denaturation requires robust pow-er-demanding instrumentation. This study addresses both issues revised primer probe designs, modified buffers low ∆T protocols which, together, speed up on conventional instruments will allow development of robust, devices. Our approach, called "FlashPCR", also allows efficient reverse transcription part one-step RT-qPCR pro-tocol, making it universally applicable rapid

Language: Английский

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Suppression of Thrips palmi population by spray-on application of dsRNA targeting V-ATPase-B

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 280, P. 135576 - 135576

Published: Sept. 11, 2024

Language: Английский

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FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(5), P. 2773 - 2773

Published: Feb. 28, 2024

Versatility, sensitivity, and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research, as well diagnostic applications. However, point-of-care (PoC) use, traditional qPCR faces two main challenges: long run times mean results are not available half an hour or more, requisite high-temperature denaturation requires more robust power-demanding instrumentation. This study addresses both issues revises primer probe designs, modified buffers, low ∆T protocols which, together, speed up on conventional instruments will allow development of robust, devices. Our approach, called "FlashPCR", uses protocol involving 15-second at 79 °C, followed by repeated cycling 1 s °C 71 together with high Tm primers specific but simple buffers. It also allows efficient reverse transcription part one-step RT-qPCR protocol, making it universally applicable rapid research

Language: Английский

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A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications

American Journal of Bioscience and Bioengineering, Journal Year: 2024, Volume and Issue: 12(3), P. 48 - 56

Published: May 17, 2024

The rapid development of molecular biology tools in insect systematics, invasion research, evolutionary ecology and biodiversity analysis has led to faster greater progress understanding behavior biology. Efficient DNA extraction is the foremost step serves as vital foundation. Several methods have been established, which are often time-consuming labour-intensive. Here, a simple, fast, low-cost protocol for common samples was developed basing on 28 specimens 16 species (7 ants, 9 bark ambrosia beetles). new shown be feasible highly efficient by comparison with commercial kit terms yield, purity PCR sensitivity. concentration through method higher than that kit, whether ant or beetle samples. A better quality extracted via indicated A<sub>260</sub>/A<sub>280</sub> mostly ranging from 1.80 2.00. There little difference between adult nymphal insects. sensitivity using both protocols comparable. For nested PCR, amplification after two rounds yielded bright signal template methods. But primers LCO1490 HCO2198, success ratio lower (85.18%). Through BLAST, these amplicons were matched related data high identity. By combining this variable platforms such loop-mediated isothermal amplification, throughput sequencing, it could assist diagnostics, biological surveys researches.

Language: Английский

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Unearthing Lactococcus lactis and Scheffersomyeces symbionts from edible wood-boring beetle larvae as a bio-resource for industrial applications

BMC Microbiology, Journal Year: 2024, Volume and Issue: 24(1)

Published: July 30, 2024

Abstract Background Gut microbiota have several advantages in influencing the host nutrition, metabolism, immunity and growth. However, understanding of gut key edible wood-boring beetle larvae remain largely undefined. In present study, characteristics two species ( Titocerus jaspideus Passalus punctiger ) from indigenous forested areas were investigated. Results Over 50% Amplicon Sequence Variants (ASVs) constituted Firmicutes T. . The dominant phyla both Bacteroidota (4.20–19.79%) Proteobacteria (15.10–23.90%). Lactococcus lactis was most abundant core prokaryote guts fungi identified insects belong to phylum Obazoa (66%) Ascomycota (> 15%). Scheffersomyeces sp. eukaryote recorded. diversity insect did not vary significantly. Most prokaryotic genes expressed predominantly associated with biosynthesis metabolism. Conclusion Our findings demonstrated that are microbes wood boring desirable probiotic properties promising use food product fermentation for improved growth performance, barrier health, intestinal flora balance immune protection human animals. Further studies highlight latest medical-based applications L. as live-delivery vector administration therapeutics against communicable non-communicable diseases warranted.

Language: Английский

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