It is important to detect specific genes expressed in the guard cells, which control gas exchange and play key roles in response to drought and salt stresses. Due to the genetic transformation of Chinese cabbage (Brassica rapa) has not been well developed, in situ RT-PCR is a valuable option for detecting guard cell specific genes. We reported an optimized protocol of in situ RT-PCR by using an FAMA homologous gene Bra001929 in Brassica rapa. FAMA in Arabidopsis has been verified to be specially expressed in guard cells. We designed specific RT-PCR primers and optimized the protocol in terms of the (a) reverse transcription time, (b) blocking time, (c) antigen-antibody incubation time, and (d) washing temperature. Our approach provides a sensitive and effective in situ RT-PCR method for locating expression in the guard cells in Brassica rapa. Moreover, we proved the guard cell specific expression of Bra001929 in the epidermis indicating its’ applicability as a marker gene for guard cells of Brassica rapa.

重要的是检测保卫细胞中表达的特定基因，这些基因控制气体交换并在应对干旱和盐胁迫中起关键作用。由于大白菜( Brassica rapa )的遗传转化尚未得到很好的开发，原位RT-PCR是检测保卫细胞特异性基因的一种有价值的选择。我们报告了在芸苔中使用FAMA同源基因Bra001929的原位RT-PCR 优化方案。拟南芥中的FAMA已被证实在保卫细胞中特别表达。我们设计了特定的 RT-PCR 引物，并在 (a) 逆转录时间、(b) 封闭时间、(c) 抗原-抗体孵育时间和 (d) 洗涤温度方面优化了方案。我们的方法提供了一种灵敏且有效的原位RT-PCR 方法，用于定位芸苔保卫细胞中的表达。此外，我们证明了Bra001929在表皮中的保卫细胞特异性表达，表明其作为芸苔保卫细胞标记基因的适用性。