Engineered plants have been widely produced for fundamental and practical use. Several methods have been developed for genetically modified crop detection and quantification; however; they still laborious and expensive. Efforts are needed to set-up diagnosis-oriented techniques as alternatives to overcome DNA extraction which remains a tedious and time-consuming procedure. Here, we established a standard direct PCR workflow using a regular Taq polymerase without prior DNA purification over a wide range of plant species. Only a small amount of fresh tissue allowed direct amplification of target gene sequences. Evaluation of accuracy, sensitivity, and reproducibility of direct PCR assay was investigated for proof-of-concept, and subsequently applied to gene detection assays and rapid transgenic revealing. The newly established method achieved full success and has amplified constitutive housekeeping genes from several plant specimens in a reproducible manner with high-quality sequencing profiles. In our case, the screening of transgenic plants confirmed that both the gfp-ER reporter gene and the npt II selectable marker were integrated into the plant genome. This direct PCR approach provides a powerful tool for large-scale PCR-based gene detection making DNA purification irrelevant. It could be easily implemented for downstream applications in the field of genetic fingerprinting, plant biotechnology, and functional genomics.

工程植物已被广泛生产用于基础和实际用途。已经开发了几种用于转基因作物检测和量化的方法；然而; 他们仍然费力和昂贵。需要努力建立面向诊断的技术作为替代方法来克服 DNA 提取，这仍然是一个乏味和耗时的过程。在这里，我们使用常规 Taq 聚合酶建立了标准的直接 PCR 工作流程，无需事先对多种植物物种进行 DNA 纯化。只有少量新鲜组织允许直接扩增目标基因序列。对直接 PCR 测定的准确性、灵敏度和再现性的评估进行了概念验证，随后应用于基因检测测定和快速转基因揭示。新建立的方法取得了全面成功，并以可重复的方式从几个植物标本中扩增了组成型看家基因，并获得了高质量的测序图谱。在我们的案例中，转基因植物的筛选证实gfp-ER报告基因和npt II 选择标记被整合到植物基因组中。这种直接 PCR 方法为基于 PCR 的大规模基因检测提供了强大的工具，使 DNA 纯化变得无关紧要。它可以很容易地用于基因指纹、植物生物技术和功能基因组学领域的下游应用。