Full
Outline
To establish a method for identification of gluten allergens in food by DNA barcoding technology.
The 4 kinds of molecular markers (psbA-trnH, trnl, rbcl and ITS2) were used to identify 5 gluten allergen species, including wheat, barley, oat, rye and buckwheat. The 4 kinds of gene sequences of each species were obtained from GenBank, and sequence alignment, genetic distance calculation and phylogenetic tree construction were performed using MEGA 11 software.
Among the 4 kinds of gene sequences, only the minimum interspecific genetic distance of the trnl sequence was greater than the maximum intraspecific genetic distance, showing a significant barcoding gap. The analysis results of the phylogenetic trees constructed by the neighbor joining method showed that the phylogenetic trees constructed based on trnl and rbcl sequences had clear branching structures, and different species could be accurately clustered. Therefore, the trnl sequence was the optimal DNA barcode for identification of the 5 gluten allergen species. Combining the trnl barcoding detection and high-throughput sequencing technology for the detection of 7 actual samples allows for simultaneous detection of multiple allergen components.
The trnl sequence, as a DNA barcode, can accurately identify gluten allergen components in food.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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