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2. Red region excitation spectra of protochlorophyllide in dark-grown leaves from plant species with different proportions of its spectral forms

Creator:
Amirjani, M. R. and Sundqvist, C.
Format:
bez média and svazek
Type:
model:article and TEXT
Subject:
chlorophyllide, etioplast, fluorescence spectrum, mutants, Pisum, species differences, Triticum, and Zea
Language:
Multiple languages
Description:
Etiolated leaves of three different species, maize, wheat, and pea, as well as a pea mutant (lip1) were used to compare the excitation spectra of protochlorophyllide (Pchlide) in the red region. The species used have different composition of short-wavelength and long-wavelength Pchlide forms. The relation between different forms was furthermore changed through incubating the leaves in 5-aminolevulinic acid (ALA), which caused an accumulation of short-wavelength Pchlide forms, as shown by changes in absorption and fluorescence spectra. This is the first time a comprehensive comparison is made between excitation spectra from different species covering an emission wavelength range of 675-750 nm using fluorescence equipment with electronic compensation for the variations in excitation irradiance. The different forms of Pchlide having excitations peaks at 628, 632, 637, 650, and 672 nm could be best measured at 675, 700, 710, 725, and 750 nm, respectively. Measuring emission at wavelengths between 675-710 nm gave an exaggeration of the short-wavelength forms and measuring at longer wavelengths gave for the pea leaves an exaggeration of the 672 nm peak. In general, an energy transfer from short-wavelength Pchlide forms to long-wavelength Pchlide forms occurred, but such an energy transfer sometimes seemed to be limited as a result of a discrete location of the Pchlide spectral forms. The excitation spectra resembling the absorption spectrum most were measured at an emission wavelength of 740 nm. Measuring the excitation at 710 nm gave higher intensity of the spectra but the short-wavelength forms were accentuated. and M. R. Amirjani, C. Sundqvist.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public

3. The effect of cross-linking of the subunits of NADPH-protochlorophyllide oxidoreductase on the aggregational state of protochlorophyllide

Creator:
Wiktorsson, B., Engdahl, S., Zhong, L.B., Böddi, B., Ryberg, M., and Sundqvist, C.
Format:
print
Type:
model:internalpart and TEXT
Language:
Multiple languages
Description:
Prolamellar bodies (PLBs) isolated from dark-grown, 6.5-d-old leaves of wheat (Triticum aesíivum L. cv. Kosack) were treated with the carboxylic acid cross-linker l-ethyl-3-[3-(diniethylaniino)propyl]carbodiimide (EDC) or with the lysině specific cross-linker 2-iniinothiolane. SDS-PAGE showed that the most prominenent cross- linked product was a dimer of the NADPH-protochlorophyllide oxidoreductase (PCR), but also larger aggregates of the polypeptide were identified by inununological detection on electro-blots. A two-dimensional diagonál gel showed that much of the cross-linking was between the PCR polypeptides. The cross-linkers induced a shift of the fluorescence peak to shorter wavelengths, a bandwidth increase of the fluorescence peak, and an increase of the fluorescence yield. In the presence of NADPH the blue shift was reduced, but the increase in the fluorescence yield still occmred. A cross-linker treatment of PLBs prior to solubilization with 1-0-n-octyl-P -D-glucopyranoside (octylglucoside) delayed, but did not prevent the spectral shifts from 657 to 646 nm and from 646 to 635 nm observed in non-cross-linked detergent- treated PLBs. The cross-linking did not prevent a spectral shift, corresponding to the Shibata shift, of Chlide. Thus the spectral shifts are not strictly coupled to disaggregation of the PCR polypeptides.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public