Real-time imaging of photosynthetic oxygen evolution from spinach using LSI-based biosensor

Shigenobu Kasai,Atsushi Suda,Yamato Sugiura,Tomohiro Honmo,Teruya Sato,Kumi Y Inoue,Kosuke Ino,Pavel Pospíšil,Masahki Matsudaira,Tomokazu Matsue,Ankush Prasad,Ryota Kunikata,Yoko Furubayashi,Yuka Hashi,Aditya Kumar

doi:10.1038/s41598-019-48561-y

Shigenobu Kasai, Atsushi Suda + Show 13 more

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https://doi.org/10.1038/s41598-019-48561-y

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The light-driven splitting of water to oxygen (O2) is catalyzed by a protein-bound tetra-manganese penta-oxygen calcium (Mn4O5Ca) cluster in Photosystem II. In the current study, we used a large-scale integration (LSI)-based amperometric sensor array system, designated Bio-LSI, to perform two-dimensional imaging of light-induced O2 evolution from spinach leaves. The employed Bio-LSI chip consists of 400 sensor electrodes with a pitch of 250 μm for fast electrochemical imaging. Spinach leaves were illuminated to varying intensities of white light (400–700 nm) which induced oxygen evolution and subsequent electrochemical images were collected using the Bio-LSI chip. Bio-LSI images clearly showed the dose-dependent effects of the light-induced oxygen release from spinach leaves which was then significantly suppressed in the presence of urea-type herbicide 3-(3,4-dichlorophenyl)−1,1-dimethylurea (DCMU). Our results clearly suggest that light-induced oxygen evolution can be monitored using the chip and suggesting that the Bio-LSI is a promising tool for real-time imaging. To the best of our knowledge, this report is the first to describe electrochemical imaging of light-induced O2 evolution using LSI-based amperometric sensors in plants.

Highlights

The light-driven splitting of water to oxygen (O2) is catalyzed by a protein-bound tetra-manganese penta-oxygen calcium (Mn4O5Ca) cluster in Photosystem II
The spatial distribution of O2 was monitored as O2 reduction current measured with the Bio-largescale integration (LSI) chip applied with −0.5 V vs. Ag/AgCl
The electrodes close with the leaf surface showed a change in reduction current in the range of −0.5 nA to ~−3.5 nA, while the electrodes far from the leaf did not show any significant changes in the reduction current

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The light-driven splitting of water to oxygen (O2) is catalyzed by a protein-bound tetra-manganese penta-oxygen calcium (Mn4O5Ca) cluster in Photosystem II. We used a largescale integration (LSI)-based amperometric sensor array system, designated Bio-LSI, to perform two-dimensional imaging of light-induced O2 evolution from spinach leaves. Spinach leaves were illuminated to varying intensities of white light (400–700 nm) which induced oxygen evolution and subsequent electrochemical images were collected using the Bio-LSI chip. To the best of our knowledge, this report is the first to describe electrochemical imaging of light-induced O2 evolution using LSI-based amperometric sensors in plants. We used our recently developed LSI-based amperometric sensor array system referred to as Bio-LSI25,26 for two-dimensional imaging of light-induced O2 evolution from spinach leaves. To the best of our knowledge, this is the first study to describe real-time electrochemical imaging of light-induced oxygen release from a photosynthetic organism using LSI-based amperometric sensors

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