Analysis, Application, and Verification of a Genetically Modified Seed Purity Determination Model Based on Binomial Distribution and F Distribution
LIU Yan-Lai1, LI Zi-He1, YANG Zhan-Sen1, HAN Tian-Yi1, REN Xue-Zhen2, JIN Fang2, YI Hong-Mei3, GAO Hong-Fei4, WU Gang4, CHENG Nan1,*, JIN Shi-Qiao2,*
1 College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; 2 National Agro-Tech Extension and Service Center, Beijing 100125, China; 3 Maize Research Center, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China; 4 Oil Crops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs/Inspection and Testing Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs/Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Abstract:With the regular cultivation of genetically modified crops worldwide and the rapid development of genetically modified seeds, some international organizations and relevant countries have established standards for the purity detection of genetically modified seeds. However, the mathematical and logical analysis of the detection schemes in these standards is still insufficient. This study aimed to provide theoretical support and design guidance for understanding and using detection schemes based on binomial distribution and F-distribution through systematic formula derivation, model analysis, application and verification. This study first started from the sources of risks in the experimental design, analyzed the key factors affecting the detection of seed transgenic purity, and proposed solutions. Secondly, a mathematical model was constructed based on the binomial distribution to quantify producer risk and user risk, and these risks were evaluated through operation curves. The third briefly described the principles of single-step and dual-step schemes. Through the analysis of mathematical models and operation curves, this study quantified the producer risk and user risk in the purity detection of genetically modified seeds, and deduced calculation formulas applicable to different detection scenarios. The scientificity and applicability of this mathematical model were verified through representative cases of germination detection of genetically modified corn and herbicide-tolerant seeds. This study systematically demonstrated the scientific nature of the mathematical model for seed transgenic purity detection based on binomial distribution and F-distribution, enhanced the transparency and reproducibility of transgenic seed purity detection methods, facilitated the correct understanding and application of standards by detection personnel. This study provides mathematical support for the formulation and revision of relevant detection norms in the future.
刘燕来, 李梓赫, 杨湛森, 韩天意, 任雪贞, 晋芳, 易红梅, 高鸿飞, 吴刚, 程楠, 金石桥. 基于二项分布和F分布的种子转基因纯度测定模型分析、应用及验证[J]. 农业生物技术学报, 2026, 34(2): 407-419.
LIU Yan-Lai, LI Zi-He, YANG Zhan-Sen, HAN Tian-Yi, REN Xue-Zhen, JIN Fang, YI Hong-Mei, GAO Hong-Fei, WU Gang, CHENG Nan, JIN Shi-Qiao. Analysis, Application, and Verification of a Genetically Modified Seed Purity Determination Model Based on Binomial Distribution and F Distribution. 农业生物技术学报, 2026, 34(2): 407-419.
[1] 金石桥, 张英, 高鸿飞, 等. 2025. GB/T 3543.12-2025, 农作物种子检验规程第12部分:品种质量转基因种子测定[S]. 北京: 国家标准化管理委员会, 国家市场监督管理总局. (Jin S Q, Zhang Y, Gao H F, et al.2025. GB/T 3543.12-2025, Rules for agricultural seed testing—Part12: Varietal quality—GMO seed testing[S]. Beijing: Standardization Administration of the People's Republic of China, State Administration for Market Regulation. [2] 李允静, 任雪贞, 肖芳, 等. 2024. 基于二重实时荧光PCR方法精准快速鉴定大豆转基因种子转化体纯度[J]. 中国农业科学, 57(23): 4698-4711. [3] (Li Y J, Ren X Z, Xiao F, et al.2024. Accurate and rapid identification of event purity for transgenic soybean seeds based on duplex real-time fluorescence PCR method[J]. Scientia Agricultura Sinica, 57(23): 4698-4711.) [4] 任佳丽. 2021. 转基因玉米标准物质-质粒阳性物质构建与检测方法的建立[D]. 硕士学位论文, 新疆农业大学, 导师: 杨卫君, 王凤格, pp. 15-27. (Ren J L.2021. Construction of transgenic maize standard substance plasmid positive substance and establishment of detection method for transgenic maize[D]. Thesis for M. S., Xinjiang Agricultural University, Supervisor: Yang W J, Wang F G, pp. 15-27.) [5] 赵新, 刘双, 刘娜, 等. 2022. 耐除草剂大豆‘DBN9004’精准定量检测方法的建立. 农业生物技术学报, 30(12): 2446-2455. (Zhao X, Liu S, Liu N, et al.2022. Establishment of accurate quantitative detection method for herbicidetolerant soybean (Glycine max) 'DBN9004'. Journal of Agricultural Biotechnology, 30(12): 2446-2455.) [6] Ahmed F E.2002. Detection of genetically modified organisms in foods[J]. Trends in Biotechnology, 20(5): 215-223. [7] Akiyama H, Sakata K, Kondo K, et al.2008. Individual detection of genetically modified maize varieties in non-identity-preserved maize samples[J]. Journal of Agricultural and Food Chemistry, 56(6): 1977-1983. [8] Cheng N, Shang Y, Xu Y C, et al.2017. On-site detection of stacked genetically modified soybean based on event-specific TM-LAMP and a DNAzyme-lateral flow biosensor[J]. Biosensors and Bioelectronics, 91: 408-416. [9] European Parliament and the Council.2001. Directive 2001/18/EC, On the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC[S]. Luxembourg: Official Journal of the European Union. [10] Grohmann L, Belter A, Speck B, et al.2014. Collaborative trial validation of a testing plan for detection of low level presence of genetically modified seeds[J]. Seed Science and Technology, 42(3): 414-432. [11] Guo M R, Xia Y M, Chen F S, et al.2023. Development of an efficient dye-based qPCR system still functional for low levels of transgenic DNA in food products[J]. Food Analytical Methods, 16(2): 445-458. [12] Hu T T, Zheng K L, Su P, et al.2020. Comparative study on protein quantitation by digital PCR with G2-EPSPS as an example[J]. Microchemical Journal, 157: 104954. [13] International Organization for Standardization.2006. ISO 24276:2006, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and derived products — General requirements and definitions[S]. Switzerland: International Organization for Standardization. [14] International Organization for Standardization.2022. ISO 22753:2021, Molecular biomarker analysis - Method for the statistical evaluation of analytical results obtained in testing sub-sampled groups of genetically modified seeds and grains general requirements[S]. Switzerland: International Organization for Standardization. [15] International Seed Testing Association.2025. ISTA Rules 2025, International rules for seed testing[S]. Berlin: International Organization for Standardization. [16] Kobilinsky A, Bertheau Y.2005. Minimum cost acceptance sampling plans for grain control, with application to GMO detection[J]. Chemometrics and Intelligent Laboratory Systems, 75(2): 189-200. [17] Kumar R, Singh C K, Kamle S, et al.2010. Development of nanocolloidal gold based immunochromatographic assay for rapid detection of transgenic vegetative insecticidal protein in genetically modified crops[J]. Food Chemistry, 122(4): 1298-1303. [18] Laffont J L, Remund K M, Wright D, et al.2005. Testing for adventitious presence of transgenic material in conventional seed or grain lots using quantitative laboratory methods: Statistical procedures and their implementation[J]. Seed Science Research, 15(3): 197-204. [19] Liu W X, Meng L X, Liu X R, et al.2022. Establishment of an ELISA method for quantitative detection of PAT/pat in GM crops[J]. Agriculture, 12(9): 1400. [20] Mano J, Yanaka Y, Ikezu Y, et al.2011. Practicable group testing method to evaluate weight/weight GMO content in maize grains[J]. Journal of Agricultural and Food Chemistry, 59(13): 6856-6863. [21] Melo L F de, Fagioli M, Sá M E de.2013. Alternative methods for detecting soybean seeds genetically modified for resistance to herbicide glyphosate[J]. Journal of Seed Science, 35: 381-386. [22] Ministério da Agricultura, Pecuária e Abastecimento (MAPA). 2006. Instrução Normativa nº 43, de 1º de dezembro de 2006—Regulamento para extensão de escopo de credenciamento dos laboratórios de análise de sementes: assegurado o limite de 1% de presença adventícia de sementes GM em sementes de algodão convencional[S]. Brasília: Ministério da Agricultura, Pecuária e Abastecimento. [23] Remund K M, Dixon D A, Wright D L, et al.2001. Statistical considerations in seed purity testing for transgenic traits[J]. Seed Science Research, 11(2): 101-120. [24] Secretaría de Agricultura, Ganadería, Pesca y Alimentación (SAGPyA). 2001. Resolución 525/2001, Tolerancias de pureza genética en semillas de cultivares de maíz (Anexo)[S]. Buenos Aires: Secretaría de Agricultura, Ganadería, Pesca y Alimentación. [25] Sub-Committee for Method Development of the German National and Federal Länder Joint Committee on Genetic Engineering.2006. Concept for seed analysis for genetically modified plant content[S]. Berlin: Joint Committee on Genetic Engineering (LAG). [26] Székács A, Lauber É, Takács E, et al.2010. Detection of Cry1Ab toxin in the leaves of MON 810 transgenic maize[J]. Analytical and Bioanalytical Chemistry, 396(6): 2203-2211. [27] United States Department of Agriculture, Agricultural Marketing Service.2025. 7 CFR Part 66, National Bioengineered Food Disclosure Standard[S]. Washington, DC: United States Department of Agriculture. [28] Zeng H J, Wang J B, Jia J W, et al.2021. Development of a lateral flow test strip for simultaneous detection of BT-Cry1Ab, BT-Cry1Ac and CP4 EPSPS proteins in genetically modified crops[J]. Food Chemistry, 335: 127627. [29] Zhang Q, Wang W R, Yang Z S, et al.2021. A portable 3D-printed biosensing device for rapid detection of genetically modified maize MON810[J]. Sensors and Actuators B: Chemical, 349: 130748.