PRACTICAL 8: Photosynthesis: Reduction of DCPIP Dye by Chloroplast

OBJECTIVES:
1. To identify the chloroplast pigment present in green plants.
2. To determine the reaction of light spectra of a plant by reducing compounds
(DCPIP).

INTRODUCTION:
2,6-dichlorophenolindophenol, ( DCPIP, also DPIP ) is a chemical compound used as
a redox dye. Oxidized, DCPIP is blue with a maximal absorption at 600 nm;
when reduced, DCPIP is colorless. DCPIP can be used to measure the rate
of photosynthesis. It is part of the Hill reagents family. When exposed to light in a
photosynthetic system, the dye is decolourised by chemical reduction. DCPIP has a
higher affinity for electrons than ferredoxin and the photosynthetic electron transport
chain can reduce DCPIP as a substitute for NADP
+
, that is normally the final electron
carrier in photosynthesis. As DCPIP is reduced and becomes colorless, the resultant
increase in light transmittance can be measured using aspectrophotometer.
For the reduction of carbon dioxide to carbohydrate there must be a source of electrons.
In the cell, NADP is the electron acceptor which is reduced in the light-dependent
reactions, and which provides electrons and hydrogen for the light-independent
reactions.
In this investigation, DCPIP (2,6-dichlorophenol-indophenol), a blue dye, acts as an
electron acceptor and becomes colourless when reduced, allowing any reducing agent
produced by the chloroplasts to be detected. The light-dependent reactions produce a
reducing agent. This normally reduces NADP, but in this experiment the electrons are
accepted by the blue dye DCPIP. Reduced DCPIP is colourless. The loss of colour in
the DCPIP is due to reducing agent produced by light-dependent reactions in the
extracted chloroplasts.






BGY 3301
PLANT PHYSIOLOGY

LAB REPORT

NAME : AZLINI BT RAZALI (169980)
MADIHAH BT ALI (170286)
NAZURA ATIKAH BT SADIR (169944)
DATE : 22
nd
APRIL 2014
PRACTICAL : 08- Photosynthesis: Reduction of DCPIP Dye by Chloroplast
LECTURER : Dr. ROSIMAH NULIT
DEMONSTRATOR : MISS SYAZREEN