Hill reaction of photosynthesis was discovered by Robin Hill (1937).

He found that isolated

chloroplasts from plants can release oxygen when they are illuminated by sunlight in the presence of
a suitable electron acceptor such as ferricyanide. To demonstrate the Hill reaction in the
laboratory dichlorophenolindophenol (DCPIP) was used as the terminal electron acceptor, replacing
NADP which was not available due to the extraction of the chloroplasts from the plant cellular
environment. The ferrocyanide is reduced (just as NADP is when the chloroplast is in vivo) while
water (H2O) is oxidized into oxygen (O2) and hydrogen cations during the reaction. By using this
technique to observe the reaction, Hill revealed a variety of valuable facts about photosynthesis. The
Hill reaction confirms that oxygen (O2) is produced by a reaction that is separate from carbon dioxide
(CO2) fixation. The reaction in which oxygen is released requires light; therefore the Hill reaction can
be described as the light dependent reaction of photosynthesis. The reaction in which oxygen is
released takes place within the chloroplast of plants and the Hill reaction demonstrated that the
release of oxygen (O2) is only a partial reaction which requires one step of photosynthesis.

Isolated chloroplasts from spinach leaves, viewed under light microscope

The Hill reaction implies that the light dependent reaction of photosynthesis is a result of a series
of redox reactions and a suitable terminal electron acceptor is required for that reaction to occur.
Plants have natural electron acceptors such as NADP, that play a major role in the oxidation of
water. The Hill reaction also shows that the natural electron acceptors of the reaction can be
substituted by an artificial electron acceptor such as DCPIP, again allowing for the discharge of
oxygen. The technique of replacing the natural electron acceptors with artificial electron acceptors
can be used in the laboratory to provide a means to measurephosphorylation in chloroplasts and
observe its relation to the discharge of oxygen in this reaction.
Hill's finding was that the origin of oxygen in photosynthesis is water (H 2O) not carbon dioxide (CO2)
as previously believed. Hill's reaction also demonstrates that the light dependent redox reaction is
the first reaction to take place in photosynthesis. The electrons thus freed by the splitting of water
provides the electrons needed by the antenna system of photosystem II where they are boosted in
energy and then sent along the electron transport system (ETS). Those high energy electrons are

able to move against a chemical potential gradient. Hence, solar energy is converted to chemical
energy by the reduction of NADP to NADPH.[4]