Fluorescence excitation spectra of different protochlorophyll(ide) (PChlide) a forms in intact etiolated cucumber cotyledons showed a pronounced increase in intensity of the 390 nm band in comparison with spectra of the pigment in Solutions. The native PChlide F657 also had an additional band at 360 nm in its excitation spectra, which disappeared after leaf homogenization and was restored by addition of dithionite. The restoration correlated with changes around 340 nm in the excitation spectrum of blue-green fluorescence. In consideration of this fact, the 360 nm band seems to correspond to energy transfer from NADPH to PChlide a. After the onset of irradiation a new band appeared at 380 nm in the excitation spectrum of the main pigment fluorescence, which was very similar to that found in the absorption spectinm of isolated Chl a. Moreover, in the Soret region the excitation spectrum had a double band splitting which disappeared only after the Shibata shift, simultaneously with the disappearance of the 360 nm band. Besides the well-known PChl(ide) F633, F642, and F657 forms, etiolated cucumber cotyledons háve emission at 673 nm. Its excitation spectrum was hardly distinguishable from the PChlide F657 fluorescence excitation spectrum in etiolated samples. In the irradiated plants this band had excitation spectrum of monomeric Chl a. The intensity of band at 673 run decreased and transiently increased after PChlide photoconversion. The dynamics of these fluorescence intensity changes under inadiation showed positive correlation both with PChlide F657 and with PChl(ide) F642, indicating the short wavelength Chl a preexistence in etiolated plants and its additional formation after the onset of irradiation.
Ginkgo biloba L. is a large tree native in China with evolutionary affinities to the conifers and cycads. However unlike conifers, the gymnosperm G. biloba is not able to synthesize chlorophyll (Chl) in the dark, in spite of the presence of genes encoding subunits of light-independent protochlorophyllide oxidoreductase (DPOR) in the plastid genome. The principal aims of the present study were to investigate the presence of DPOR protein subunits (ChlL, ChlN, ChlB) as well as the key regulatory step in Chl formation: aminolevulinic acid (ALA) synthesis and abundance of the key regulatory enzyme in its synthesis: glutamyl-tRNA reductase (GluTR). In addition, functional stage of photosynthetic apparatus and assembly of pigment-protein complexes were investigated. Dark-grown, illuminated and circadian-grown G. biloba seedlings were used in our experiments. Our results clearly showed that no protein subunits of DPOR were detected irrespective of light conditions, what is consistent with the absence of Chl and Chl-binding proteins (D1, LHCI, LHCIIb) in the dark. This correlates with low ALA-synthesizing capacity and low amount of GluTR. The concentration of protochlorophyllide (Pchlide) in the dark is low and non-photoactive form (Pchlide633) was predominant. Plastids were developed as typical etioplasts with prollamelar body and few prothylakoid membranes. Continual illumination (24 h) only slightly stimulated ALA and Chl synthesis, although Pchlide content was reduced. Prollamelar bodies disappeared, but no grana were formed, what was consistent with the absence of D1, LHCI, LHCIIb proteins. Lightinduced development of photosynthetic apparatus is extremely slow, as indicated by Chl fluorescence and gas exchange measurements. Even after 72 h of continuous illumination, the values of maximum (Fv/Fm) and effective quantum yield (ΦPSII) and rate of net photosynthesis (PN) did not reach the values comparable with circadian-grown plants. and A. Pavlovič ... [et al.].
In contrast to angiosperms, which accumulate protochlorophyllide after application of aminolevulinic acid in the dark, feeding with aminolevulinic acid (0.01-20 mM) via the roots in the 18-d-old seedlings of Norway spruce (Picea abies) stimulated not only protochlorophyllide but also chlorophyll accumulation. and A. Pavlovič ... [et al.].
Life and research results of Pavel Siffel, a talented but untimely deceased Czech scientist in photosynthesis, are reviewed. He studied biophysics and physiology of chlorophyll, its complexes with proteins, their absorption and fluorescence spectra, activities in mutants and transformants, dealt with chlorophyll biosynthesis and protochlorophyllide photoreduction, pigments in plants grown at CO2 deficiency and under simulated acid rain, with changes accompanying leaf and plant development, photobleaching, etc. He participated in construction of specialised spectrofluorometers, finally he built the kinetic spectrophotometer SpeKin. and J. Květoň ... [et al.].
Charactenstícs of the pigment apparatus were studied in plants grown from tubers of the oř 3’’^ doně generation of two cultivars of potato (Solanum tuberosum L) transformed with Agrobacterium tumefaciens strains. The cv. Zvíkov carries pRiA4b T-DNA of A. rhizogenes and the vector plasmid pGA472 T-DNA carrying the gene for kanamycin resistance. The cv. Kamýk carries the vector plasmid pCB1339 which contains the kanamycin resistance gene and the cytokinin synthesis gene. In the field experiments, the plant dry matter, the relative water content, the chlorophyll (Chl) a and b contents, the Chl a/b ratio and the course of slow Chl fluorescence induction curves were not signifícantly different in transformed and control plants. The amoímt of the Chl-protein complexes of the reaction centres of both photosystems separated by electrophoresis was similar for both transformed and control plants. Howevér, the content of oligomeric light-harvesting complex was significantly higher in both the transformants.
A trace amount of chlorophyll (Chl) a was found in addition to protochlorophyllide (PChlide) a and protochlorophylls (PChls) a in acetone extracts of etiolated cucumber cotyledons by HPLC. There was, however, no chlorophyllide (Chlide) a, which was present in extracts from illuminated cotyledons. After 10 min exposure of cotyledons to light, all PChlide a was photoreduced to Chlide a. Hence, the whole pool of PChlide seems to be photoactive. The "photoinactive" spectral form PChl(ide) F633 was identified as a mixture of monovinyl-PChls a esterified by different alcohols.