In-depth analysis of the endogenous reference genes used in the quantitative PCR detection systems for rice

文献类型： 外文期刊

第一作者： Zhang, Li

作者： Zhang, Li;Cao, Yinglong;Wu, Gang;Wu, Yuhua;Lu, Changming;Liu, Xin

作者机构：

关键词： Oryza sativa;Endogenous reference gene;Qualitative PCR;Real-time quantitative PCR;Genetically modified organism detection

期刊名称：EUROPEAN FOOD RESEARCH AND TECHNOLOGY （ 影响因子：2.998； 五年影响因子：3.005 ）

ISSN： 1438-2377

年卷期： 2012 年 234 卷 6 期

页码：

收录情况： SCI

摘要： To standardize the rice-specific PCR detection methods, five previously reported rice () taxon-specific genes were compared and evaluated. The investigated genes included the rice - gene (), the gene -, the gene (), the gene () and the gene (). Sequencing analyses revealed that among the tested rice cultivars, single-nucleotide polymorphisms (SNPs) existed in the , , - and amplicons, though no statistically significant effect on their Ct values was found. The - and systems were found to produce amplicons in non-rice species, such as sugarcane and broomcorn. The quantitative real-time PCR results revealed that this cross-reaction led to an underestimate of the GM (genetically modified) rice content. With the exception of the aforementioned shortcomings, these five endogenous reference genes all have acceptable amplification efficiencies, which ranged from 98 to 108 %, and high sensitivity within the limit of detection (LOD) values, which ranged from 5 to 10 copies of the haploid genome. In estimating the GM content in blinded rice samples, these five systems produce relatively accurate quantitative results with deviations less than 15 %, but the system produced the most accurate quantitative results. Therefore, we have determined that the gene is the most suitable rice reference gene, and the and genes can also be used as rice reference genes because they have good commutability with the gene. Care should be taken when interpreting results based on the and - genes owing to their cross-reaction with other species.

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