An optimized high-quality DNA isolation protocol for spodoptera frugiperda J. E. smith (Lepidoptera: Noctuidae)

Abstract

An optimized high-quality DNA isolation protocol was developed using body segment tissue from the Fall Armyworm (Spodoptera frugiperda), that will allow documenting genetic variability based on biotypes, facilitating studies on the appearance, distribution and population dynamics of the fall armyworm at the molecular level. The resulting protocol is an easy-to-use, timesaving method that can rapidly achieve high quality, high-yielding total genomic DNA, using chemicals and everyday consumables available in a molecular laboratory. This new method of DNA extraction avoids the contamination of polysaccharides, salts, phenols, proteins and other cellular by-products that can interfere with subsequent reactions. DNA purity estimates reveal A260: A280 ratios greater than 1.9, which were evidenced by quality test on agarose gel, observing complete integrity and high purity of the resulting samples, and yielded 30–99 µg/g of total DNA. Therefore, the quality of the DNA produced from this extraction is suitable for subsequent molecular applications: (i) next generation whole genome sequencing, (ii) conventional polymerase chain reaction for genotyping, (iii) barcodes and (iv) gene cloning. In addition, to become an anticipating diagnostic tool for invasive lepidopteran larval stages: The resulting protocol is an easy-to-use time-saving method. This new extraction method prevents contamination from polysaccharides, salts, phenols, proteins, and other cellular sub-products. DNA purity estimations reveal A260:A280 ratios above 1.9.

Impact of Bt corn expressing Bacillus thuringiensis Berliner insecticidal proteins on the growth and survival of Spodoptera frugiperda larvae in Colombia

“…This information highlights the importance of constant monitoring (under field conditions) to detect variations in the susceptibility of S. frugiperda populations, and this is particularly important in tropical conditions. In addition, careful attention needs to be paid to the high genetic variability of S. frugiperda currently reported in Colombia ( 6 , 30 , 60 ). This monitoring also needs to be carried out to compare results obtained in laboratory conditions and the efficacy of technologies used in the field ( 26 , 29 ).…”

“…Fall armyworm, Spodoptera frugiperda , is one of the most serious pests in agricultural production in the Americas ( 1 , 2 ). It is distributed widely among hosts in the Western Hemisphere (in more than 27 plant families), it can migrate very easily ( 3 – 6 ), and has had a big economic impact on corn, sorghum, rice, and cotton crops ( 2 , 3 , 7 , 8 ). S. frugiperda was first reported in West Africa in 2016 ( 2 , 9 , 10 ) and then spread throughout much of sub-Saharan Africa by 2017 ( 11 ).…”

Molecular characterization of Insect Parasitic nematodes, and assonated Symbiotic bacteria as Ecotoxic effect of Invasive insect larvae

“…Approximately 1000-2000 sterilized IJs from each community were used independently for rDNA extraction (Phan et al, 2001). To put to the test the insect South American tomato pinworm Tuta absoluta rDNA extraction procedure provided by (Marin et al, 2021).…”

High-quality DNA isolation protocol for detection of Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) in standard wheat germ trap

“…Extraction of larger sample volumes from a standard wheat germ trap would require significant amounts of reagents costing upwards of $50 per extraction. In addition, commercial kits are normally not suitable for DNA extraction from samples containing high levels of polysaccharides and/or proteins [ 14 ], such as is present in wheat germ. Wheat germ polysaccharides interact with DNA to form a highly viscous solution unsuited to filtering through a DNA extraction column.…”