European larch (Larix decidua Mill.) and Norway spruce [Picea abies (L.) Karst.] synthesize chlorophyll (Chl) in darkness. This paper compares Chl accumulation in 14-d-old dark-grown seedlings of L. decidua and P. abies after shortterm (24 h) feeding with 5-aminolevulinic acid (ALA). We used two ALA concentrations (1 and 10 mM) fed to cotyledons of both species in darkness and in continuous light. The dark-grown seedlings of L. decidua accumulated Chl only in trace amounts and the seedlings remained etiolated. In contrast, P. abies seedlings grown in darkness were green and had significantly higher Chl content. After ALA feeding, higher protochlorophyllide (Pchlide) content was observed in L. decidua than in P. abies cotyledons incubated in darkness. Although short-term ALA feeding stimulated the synthesis of Pchlide, Chl content did not change significantly in cotyledons incubated in darkness. The Chl accumulation in cotyledons fed with ALA was similar to the rate of Chl accumulation in the controls. Higher Chl accumulation was reported in control samples after illumination: 86.9% in L. decidua cotyledons and 46.4% in P. abies cotyledons. The Chl content decreased and bleaching occurred in cotyledons incubated with ALA in light due to photooxidation. Analyses of Chlbinding proteins (D1 and LHCIIb) by Western blotting proved differences between Chl biosynthesis in L. decidua and P. abies seedlings in the dark and in the light. No remarkable increase was found in protein accumulation (D1 and LHCIIb) after ALA application. Our results showed interspecific difference in Chl synthesis between two gymnosperms. Shortterm ALA feeding did not stimulate Chl synthesis, thus ALA synthesis was not the rate-limiting step in Chl synthesis in the dark., N. Maximová, Ľ. Slováková., and Obsahuje bibliografii
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.].
A unified multibranched chlorophyll (Chl) biosynthetic pathway is proposed. The proposed pathway takes into account the following considerations: (a) that the earliest putative precursor of monovinyl Chl b that has been detected in higher plants is monovinyl protochlorophyllide b, (b) that in most cases, Chl b biosynthesis has its roots in the Chl a biosynthetic pathway, (c) that the Chl a biosynthetic pathway exhibits extensive biosynthetic heterogeneity, (d) that Chl biosynthesis may proceed differently at different stages of greening and in different greening groups of plants. Integration of the Chl a and b biosynthetic pathways into a unified multibranched pathway offers the functional flexibility to account for the structural and biosynthetic complexity of photosynthetic membranes. In this context, it is proposed that the unified, multibranched Chl a/b biosynthetic pathway represents the template of a Chl-protein biosynthesis center where photosystem (PS) 1, PS2, and light-harvesting Chl-protein complexes are assembled into functional photosynthetic units. The individual biosynthetic routes or groups of two to three adjacent biosynthetic routes may constitute Chl-protein biosynthesis subcenters, where specific Chl-protein complexes are assembled. and C. A. Rebeiz ... [et al.].
The photodynamic damage of the sensitive plants wheat and mustard, treated with chlorophyll (Chl) precursors 5-aminolevulinic acid (ALA) and glutamic acid (Glu) and with 1,10-phenanthroline (Phen), was caused by tetrapyrroles, which accumulated after 17 h in the dark period, followed by 12 h of irradiation with "white light". The effect of accumulated Chl in mustard plants was accompanied by changes in the amounts of the Chls and carotenoids and by dehydration of the tissues, partial chlorosis, and necrosis. The molecular nature of the specific photodynamic sensitivity of the mustard and wheat plants under the influence of Phen and Chl precursors was important: accumulation of tetrapyrroles was a necessary, but not only reason for photodynamic damage of the plants. The degree of leaf damage was related to the amount and chemical nature of accumulated tetrapyrroles and to the greening group to which the investigated plant belongs. and V. Toneva, T. Gechev, I. Minkov.
5-aminolevulinic acid (ALA) is an essential precursor for the biosynthesis of tetrapyrrols such as heme and chlorophyll (Chl). Previous studies have focused mainly on promotive effects of exogenous ALA on plant growth, while regulatory mechanisms affecting Chl biosynthesis have been only partially discussed. In the present study, the ameliorative role of exogenous ALA was investigated on Chl and endogenous ALA biosynthesis in six-day-old etiolated oilseed rape (Brassica napus L.) cotyledons during the de-etiolation stage. We showed that exogenously applied ALA of a low dosage enhanced Chl and ALA accumulation in cotyledons, while 600 µM ALA treatment inhibited the accumulation of Chl and ALA severely. However, the gene expression levels of glutamyl-tRNA reductase (HEMA) and glutamate-1-semialdehyde aminotransferase (GSA) were not affected under either low or high ALA concentrations. Furthermore, water deficit induced by polyethylene glycol 6000 (PEG) suppressed the Chl and ALA accumulation in cotyledons, while the inhibition was partially alleviated in the cotyledons pretreated with ALA. The decrease in Chl biosynthesis induced by PEG stress was assumed to be related to downregulation of HEMA and Mg-chelatase ChlH (ChlH), and upregulation of ferrochelatase (FC) genes. Moreover, exogenously applied ALA did not show any effect on the expression of Chl synthesis-related genes under the PEG treatment. These results showed a difference in suppressing ALA synthesis due to the high concentration of ALA and PEG. Exogenously applied ALA did not affect the expression of HEMA and GSA, thus exogenous ALA regulated Chl synthesis not via the regulation of transcriptional level in ALA biosynthesis. However, the inhibition in Chl and endogenous ALA accumulation by the PEG treatment may be attributed to downregulation of HEMA and ChlH, and upregulation of FC., D. Liu, D. D. Kong, X. K. Fu, B. Ali, L. Xu, W. J. Zhou., and Seznam literatury
In the present studies, we have found a fragment of amino acid sequence, called TFT motif, both in light-dependent protochlorophyllide oxidoreductase (LPOR) and in the L subunit of dark-operative (light-independent) protochlorophyllide oxidoreductases (DPOR). Amino acid residues of this motif shared similar physicochemical properties in both types of the enzymes. In the present paper, physicochemical properties of amino acid residues of this common motif, its spatial arrangement and a possible physiological role are being discussed. This is the first report when similarity between LPOR and DPOR, phylogenetically unrelated, but functionally redundant enzymes, is described., M. Gabruk ... [et al.]., and Obsahuje bibliografii
The effect of Hg++ was studied on the arrangement and photoactivity of NADPH:protochlorophyllide oxidoreductase (POR) in homogenates of dark-grown wheat (Triticum aestivum L.) leaves. 77 K fluorescence emission spectra of the homogenates were recorded before and after the irradiation of the homogenates and the spectra were deconvoluted into Gaussian components. The mercury treatment caused a precipitation of the membrane particles, which was followed by a remarkable decrease of the fluorescence yield. 10-3 M Hg++ decreased the ratio of the 655 nm-emitting protochlorophyllide (Pchlide) form to the 633 nm-emitting form. 10-2 M Hg++ shifted the short wavelength band to 629-630 nm and a 655 nm form was observed which was inactive on irradiation. This inhibition may be caused by serious alteration of the enzyme structure resulting in the trans-localisation of NADPH within the active site of POR. and K. Lenti, F. Fodor, B. Böddi.
The possibility of simultaneously ušed chlorophyll (Chl) synthesis precursor (glutamic acid) and metal chelator (2,2'-dipyridyl) as a photodynamic inhibitor of the chlorophyll synthesis was studied. Wheat {Triticum aestivum L.) and vegetable marrow {Cucurbita moschata Duch.) leaves were treated with 2,2'-dipyridyl (2,2'- DP), and 2,2'-DP along with glutamic acid which increased the protochlorophyllide (Pchlide) content in the vegetable marrow plants to a higher extent. An irradiation of the treated leaves caused an inhibition of Pchlide reduction which was more notable in those of the dicotyledonous vegetable marrow plant.