As a tremendously noxious and extensively utilized chemical reagent, hydrazine (N2H4) has become a serious threat to ecosystem and human health. Thus, it is desirable to exploit an efficient method for real-time tracking of hydrazine. Here, a novel ratiometric fluorescent probe PBQ-AB for hydrazine was rationally constructed from isolongifolanone. This probe displayed an extremely large Stokes shift of 230 nm and could selectively recognize hydrazine in the presence of other competitive species within an extremely short time ( 40 s). PBQ-AB also displayed some fascinating merits in the detection of hydrazine, including low detection limit (48 nM), wide pH range (5–12), excellent photostability (>240 min), and well-resolved emission wavelength shift (148 nm). Moreover, this probe was utilized to fabricate a ready-to-use electrospinning nanofibrous membrane for convenient detection of hydrazine vapor by virtue of smartphone. Furthermore, PBQ-AB was capable of determining hydrazine contaminant in environmental soil and water samples. Additionally, its favorable performance for detecting hydrazine was successfully demonstrated in live HeLa cells as well as in live Arabidopsis thaliana tissues, manifesting its promising application for labeling hydrazine in living systems. Therefore, we believed that this probe has great potential in environmental analysis and health supervision.