Nitric oxide is involved in stomatal development by modulating the expression of stomatal regulator genes in Arabidopsis

As sessile organisms, plants require many flexible strategies to adapt to the environment. Although some environmental signaling pathways regulating stomatal development have been identified, how stomatal regulators are modulated by internal and external signals to determine the final stomatal abundance requires further exploration. In our studies, we found that nitric oxide (NO) promotes stomatal development with increased stomatal index as well as the relative number of meristemoids and guard mother cells [%(M+GMC)] in NO-treated wild-type Arabidopsis plants; this role of NO was further verified in the nox1 mutant, which exhibits higher NO levels, and the noa1 mutant, which exhibits low NO accumulation. To gain insight into the molecular mechanisms underlying the effect of NO, we further assayed the expression of genes involved in stomatal development and found that NO induces the expression of the master regulators SPCH, MUTE and SCRM2 to initiate stomatal development. In addition, MPK6 is also involved in NO-promoted stomatal development, as MPK6 expression was repressed in nox1 and NO-treated plants, and transgenic plants overexpressing MPK6 were less sensitive to SNP treatment in terms of changes in the%(M+GMC). Thus, our study shows that NO promotes the production of stomata by up-regulating the expression of SPCH, MUTE and SCRM2 and down-regulating MPK6 expression.