This paper presents an impedimetric biosensor for monitoring of crops using novel microneedle electrodes for minimally invasive, sensitive measurements, compatible with smart agriculture developments. The biosensor was fabricated through polydimethylsiloxane imprinting using a high-resolution 3D printed negative mold. This method allows for fabrication of tailored microneedles with heights up to 500 μm, required to penetrate through the epidermis of a leaf. The sensor was tested by recording the bioimpedance of Barely leaves and comparing its performance to conventional planar and needle electrodes. Furthermore, we present evidence of a diurnal pattern in plant bioimpedance that was detected by monitoring the leaf bioimpedance under controlled conditions.