Establishment of a real-time PCR assay based on SYBR green I for detection of the gene expression of Lamiophlomis rotate (Benth.)

This study developed a real-time reverse-transcription polymerase chain reaction (RT-PCR) method based on SYBR Green I fluorescent for detection of Lamiophlomis rotate (Benth.) gene mRNA. According to the gene sequence in the conservative regions of chalcone synthase (LrCHS) and phenylalanine ammonia-lyase (LrPAL) in L. rotate, the specific primers were designed with LrCHS and LrPAL to construct real-time RT-PCR assay The standard curves showed good linear relationships with the correlation cofficient abave 0.995 in the range of 1伊105 to 1伊1011 copies/L The melting curve analysis showed that the product was specific to a single peak and no primer-dimers with high specificity and sensitivity The expression level of CHS gene in different organization of L. rotate showed flowers>leaves>petioles>roots>stems trend, and the expression level of PAL gene showed petioles>leaves>flowers>roots>stems trend. This assay could successfully provide a technical platform to research L. rotate genes at the mRNA level in the quantitative analysis.