Although the metabarcoding results were qualitatively congruent with the actual composition of the pollen mixes in most cases, the quantitative results based on read-counts did not match the actual ratio of pollen grains in the mixes. Metabarcoding analysis of the artificial pollen mixes showed that nuclear DNA barcodes ITS1, ITS2, and 5′-ETS proved to be more efficient than the plastome barcode in both amplification from pollen DNA and identification of grass species. Their capability to correctly identify grass pollen was tested on artificial pollen mixes of various complexity.
Barcode sequences were analyzed and compared by the barcoding gap and intra- and interspecific distances. The DNA extraction method for single-species pollen samples and mixes was optimized to increase the yield for amplification and sequencing of pollen DNA. New Poaceae-specific primers for 5′-ETS were designed and tested to obtain a 5′-ETS region less than 600 bp long, suitable for high-throughput sequencing. Sequences for the database were Sanger sequenced from live field and herbarium specimens and collected from GenBank.
A diverse set of 14 grass species of different genera were selected to create a local reference database of nuclear ITS1, ITS2, 5′-ETS, and plastome trnL-F DNA barcodes. One of the solutions to this problem is the metabarcoding approach employing DNA barcodes for taxonomical identification of species in a mix by high-throughput sequencing of the pollen DNA. Aerobiological monitoring is a necessary element of the complex of anti-allergic measures, but the similar pollen morphology of Poaceae species makes it challenging to discriminate species in airborne pollen mixes, which impairs the quality of aerobiological monitoring. Grass pollen is one of the major causes of allergy. In most samples, undeclared components have been detected, such as bindweed (Convolvulus) and ragweed (Ambrosia), which are known toxic and allergy-causing plants.
It was confirmed by classic methods of botanical analysis (anatomy and palynology). They have revealed the substitution of fireweed (Epilobium angustifolium L.) by Lythrum sp.
#Highc torrent torrent#
We report here a comparative study of the two most widely used sequencing platforms - Illumina and Ion Torrent - for composition analysis of herbal teas, and show that both technologies yield congruent results, both qualitatively and quantitatively.
However, the protocols of DNA barcoding applied to food analysis are not yet fully established testing and optimization are required to achieve the highest accuracy and cost efficiency. The rapid development of high-throughput sequencing (HTS) methods offers new opportunities for food quality control and identification of food components using the DNA barcoding approach (metabarcoding in cases of complex mixes).
0 kommentar(er)
