Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using ^<13>N-labeled nitrogen gas(Plant Nutrition)

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Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean (Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ^<13>N-labeled radioactive nitrogen gas (half life: 9.97min). ^<13>N was produced from a ^<16>O (p,α) ^<13>N nuclear reaction. The target chamber was filled with CO_2 and irradiated for 10min with protons at an energy of 18.3MeV and an electric current of 5μA, which was delivered from a cyclotron. All CO_2 in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35mL fraction, including the peak of [^<13>N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10mL of O_2 and 5mL of N_2 and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ^<13>N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17±0.10μmol N_2 h^<-1> from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012±0.011μmol N_2 h^<-1>. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [^<13>N]N_2 and PETIS.

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using ^<13>N-labeled nitrogen gas(Plant Nutrition)

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