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<p>Transgenic maize LD05, developed independently by the Shandong Academy of Agricultural Sciences, exhibits promising industrialization potential due to its dual traits of insect resistance and herbicide tolerance. This was achieved through the insertion of an insect-resistant gene (<i>m2cryAb-vip3A</i>) and a herbicide-tolerant gene (<i>bar</i>) into its genome. Establishing a quantitative detection method for transgenic maize LD05 and its derivatives is crucial for ensuring the safety regulation of transgenic crops. In this study, we designed specific primers and Taqan probes based on the 3′ end-to-flank sequence of transgenic maize LD05. A real-time fluorescence PCR was developed for the specific detection of LD05 maize transformants by primer screening, specificity testing, PCR system optimization, standard curve establishment, and accuracy and precision assessments. This method determines the detection limit of LD05 transgenic components is 0.05% by mass fraction, and the quantitative detection limit is 100 copies. Additionally, the method’s reliability was confirmed through inter-laboratory validation. This highly specific and accurate quantitative detection method provides robust technical support for the safety supervision of transgenic maize LD05.</p>

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Establishment of an event-specific quantitative real-time PCR method for genetically modified maize LD05 with combined insect resistance and herbicide tolerance

  • 期刊论文

  • 发表日期：

    2025年10月25日

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  • Wenlan Li,
  • Wujun Jin,
  • Mei Dong,
  • Xinwei Hou,
  • Zhaohua Ding,
  • Zhaodong Meng,
  • Runqing Yue

摘要

Transgenic maize LD05, developed independently by the Shandong Academy of Agricultural Sciences, exhibits promising industrialization potential due to its dual traits of insect resistance and herbicide tolerance. This was achieved through the insertion of an insect-resistant gene (m2cryAb-vip3A) and a herbicide-tolerant gene (bar) into its genome. Establishing a quantitative detection method for transgenic maize LD05 and its derivatives is crucial for ensuring the safety regulation of transgenic crops. In this study, we designed specific primers and Taqan probes based on the 3′ end-to-flank sequence of transgenic maize LD05. A real-time fluorescence PCR was developed for the specific detection of LD05 maize transformants by primer screening, specificity testing, PCR system optimization, standard curve establishment, and accuracy and precision assessments. This method determines the detection limit of LD05 transgenic components is 0.05% by mass fraction, and the quantitative detection limit is 100 copies. Additionally, the method’s reliability was confirmed through inter-laboratory validation. This highly specific and accurate quantitative detection method provides robust technical support for the safety supervision of transgenic maize LD05.