Nitrogen management through monitoring
of crop nitrate status can
improve agricultural productivity, profitability, and environmental
performance. Current plant nitrate test methods require expensive
instruments, time-intensive labor, and trained personnel. Frequent
monitoring of in planta nitrate levels of the stalks
in living plants can help to better understand the nitrogen cycle
and the physiological responses to environmental variations. Although
existing enzymatic electrochemical sensors provide high selectivity,
they suffer from short shelf life, high cost, low-temperature storage
requirement, and potential degradation over time. To overcome these
issues, an artificial enzyme (vitamin B12 or VB12) and a two-dimensional
material (graphene oxide or GO) are introduced into a conventional
photoresist (SU8) to form a bioresin SU8-GO-VB12 that can be patterned
with photolithography and laser-pyrolyzed into a carbon-based nanocomposite
C-GO-VB12. The electrocatalytic activity of the cobalt factor in VB12,
the surface enhancement properties of GO, and the porous feature of
pyrolytic carbon are synergized through design to provide C-GO-VB12
with
a superior ability to detect nitrate ions through redox reactions.
In addition, laser writing-based selective pyrolysis allows applying
thermal energy to target only SU8-GO-VB12 for selective pyrolysis
of the bioresin into C-GO-VB12, thus reducing the total energy input
and avoiding the thermal influence on the materials and structures
in other areas of the substrate. The C-GO-VB12 nitrate sensor demonstrates
a year-long shelf lifetime, high selectivity, and a wide dynamic range
that enables a direct nitrate test for the extracted sap of maize
stalk. For in situ monitoring of the nitrate level
and dynamic changes in living maize plants, a microelectromechanical
system-based needle sensor is formed with C-GO-VB12. The needle sensor
allows direct insertion into the plant for in situ measurement of nitrate ions under different growth environments
over time. The needle sensor represents a new method for monitoring in planta nitrate dynamics with no need for sample preparation,
thus making a significant impact in plant sciences.