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Outline
To explore the performance differences of various maize seed tissues in genome extraction and nucleic acid testing to clarify the optimal choice and its impact on clustered regularly interspaced short palindromic repeats/associated protein 12a (CRISPR/Cas12a)-based detection system.
In this study, the genome extraction efficiencies of maize seed embryo, endosperm and whole kernel were compared. Through the conventional polymerase chain reaction (PCR) cascade CRISPR/Cas12a system, the extracted products from each part were amplified and identified, and the impact of the extracted genome on subsequent nucleic acid detection was evaluated. By comparing the DNA extraction efficiency, amplification effect and fluorescence signal intensity, the adaptability of samples from various parts in the test was verified.
There were differences in the efficiency of DNA extraction from different tissues of corn seeds. The genome concentration in the embryo was the highest,followed by the whole kernel, and the endosperm was relatively low, which was in line with the biological structural characteristics of corn seeds. The genomes in each part could be successfully amplified, and the cascade CRISPR/Cas12a system produced a clear positive signal. The qualitative detection limit of the method constructed in this study was 0.1 wt%, which met the detection requirements of genetically modified ingredients in most countries and regions.
Differences in extraction efficiency are found among maize seed tissues, but all parts supports effective nucleic acid amplification and CRISPR/Cas12a detection. In practical applications, tissue separation is not required; efficient and specific detection can be achieved using whole seeds or randomly selected samples. The method developed in this study does not require large-scale instrumentation and is suitable for on-site detection of genetically modified maize seeds, demonstrating potential for application in the safety screening of genetically modified 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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