I suggest to use Fluorcam system as provided in the following link. You can measure chlorophyll fluorescence parameters. It can give you lots of information.

 http://www.psi.cz/products/fluorcams/

Hi Sasan

Hi Stephany

Many thanks for your answers my question but I asked about the details of the  method of  (2,6-dichlorophenolindophenol) DCPIP  because I can afford the kits and it is more easier and done with spectrophotometer

Hi There,

You are probably referring to the famous "hill reaction". During the 30's Hill and colleagues used DCPIP as the electron acceptor of the linear electron flow  on  thylakoid membranes of isolated chloroplasts. Whenever electron is transferred to the DCPIP, the molecule changes its color from blue to colorless. Measuring its absorbance change with time at 620 nm will reveal the rate of electron transfer through photosynthetic electron transport chain.

To find general details about the Hill reaction see this:

https://en.wikipedia.org/wiki/Photosynthesis

and a nice tribute written by Walker DA 2002:

https://www.ncbi.nlm.nih.gov/pubmed/16245102

you can find specific protocols for hill's reaction anywhere on the internet.

Good luck !

PS: please note though, DCPIP is not the terminal acceptor of the chain like NADP+, rather, it accepts the electrons from the middle of the chain.

Hello,

I always use different fluometers to measure ETR1 or ETR2. I want to know the advantages of this DCPIP method. Thank you

Hi there and thank you for reviewing my answer.

When measuring ETR with flourometers, one will get an estimate of the rate in flux units. This is due to the formulation of the ETR, which is the product of the quantum yield (relative units) multiplied by light quantity flux units and other constants such as the absorption coefficient and/or the ratio between PSI and PSII (usually it is set to  0.5 in commercial pocket size fluorometers, i.e. 1:1 PSII:PSI).

When dealing with the DCPIP method its a whole other realm because here you can get the estimate in uM/sec, i.e. conventional units. By using the DCPIP you can estimate the ETR coming out of PSII while comparing this information with oxygen evolution from PSII (also measured in uM/sec). Classic experiment will be to estimate the moles of electrons running through the chain compared to the oxygen evolved during photosynthesis. It will be interesting to see if it equals 4:1 (4 electrons per O2 molecule). This is hardly the case, but still nice experiment that tells alot about the photosynthetic apparatus.

Hope that helps,

Thank you very much.

We usually work with Maxi-Imaging-PAM, by Walz (Germany). It works really well and gives tons of valuable information about all photosynthetic process.

http://www.walz.com/products/chl_p700/imaging-pam_ms/maxi_version.html

Hi, my peers 

Thank you very much for following my question

I have found a publication related to this issue " Phosphorylation of PSII proteins in maize thylakoids in the presence of Pb ions" 

(Romanowska et al. 2012) Journal of Plant Physiology 169 (2012) 345– 352.

The measurements of PSII electron transfer activity in the presence of

(2,6-dichlorophenolindophenol) DCPIP was done as follow:

Activity of PSII was tested on  thylakoids of maize

chloroplasts, isolated from the control and treated Pb ions leaves.

Water or DPC (1,5-diphenylcarbazide) were used as an electron

donors for PSII, and DCPIP photoreduction was estimated spectrophotometrically

at 595 nm. Thylakoids (10 g Chl mL−1) were

resuspended in a buffer containing 330 mM sorbitol, 40 mM HEPES-KOH

(pH 7.6), 1 mM KH2PO4, 5 mM NaCl, 5 mM MgCl2, 5 mM NH4Cl.

DCPIP (0.1 mM) and DPC (0.5 mM) were added to the sample just

before reaction. During the reaction, samples were illuminated at

1000 mol photons m−2 s−1. The reduction of DCPIP was measured

at 30 s intervals during the 1-min assays and the measurement was

repeated five or six times. Difference between initial absorbance

and absorbance after illumination was taken to calculate PSII activity.