Abstract:

This study describes an innovative approach for detecting mRNA, called in situ reverse transcription (ISRT), which marries in situ hybridization (ISH) with reverse transcription. Specific primers were designed for mRNA-specific reverse transcription. The parameters and key steps of ISRT were optimized. Based on our experience, with proper controls ISRT, can detect mRNA signals with high sensitivity and high specificity. ISRT was used to detect the mRNA of a rice MADS-box gene, M79, which belongs to the AGL2 subgroup. As seen with conventional ISH, M79 is expressed at all stages of rice development, in both vegetative and reproductive processes. ISRT is relatively fast and easy-to-perform, suggesting that this method will become a reliable, fundamental technique that can be used to localize mRNA in situ.

Key words: M79 , MADS-box, Oryza sativa , in situ reverse transcription (ISRT), flower development