Abstract: In recent years, the real-time transcription polymerase chain reaction (RT-PCR) has been widely used to molecular diagnostics, pathological analysis and food safety test as a new methodology of nucleic acid sequences quantitative analysis. Although the RT-PCR was a quantitative accurate, highly sensitive and high-throughput technology, its study on plant gene expression was relatively few. The article summarized the applications of RT-PCR in analysis of gene integrated expression, gene families expression, gene differential expression and transgenic expression. Based on case studies, the influencing factors of gene expression detection were analyzed by including reverse transcription and housekeeping gene selection. However, the technology still needed to be improved due to its reproducibility, sensitivity and normalization flaws. With the development of molecular biotechnology, the RT-PCR will play a more extensive role in plant genetics and breeding, plant pathological quarantine and molecular evolution.