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Comparison of chlorophyll and carotenoid concentrations among Russian wheat aphid (Homoptera: Aphididae)-infested wheat isolines.

Russian wheat aphid, Diuraphis noxia (Mordvilko), feeding injury on 'Betta' wheat isolines with the Dn1 and Dn2 genes was compared by assessing chlorophyll and carotenoid concentrations, and aphid fecundity. The resistant Betta isolines (i.e., Betta-Dn1 and Betta-Dn2) supported similar numbers of aphids, but had significantly fewer than the susceptible Betta wheat, indicating these lines are resistant to aphid feeding. Diuraphis noxia feeding resulted in different responses in total chlorophyll and carotenoid concentrations among the Betta wheat isolines. The infested Betta-Dn2 plants had higher levels of chlorophylls and carotenoids in comparison with uninfested plants. In contrast, infested Betta-Dn1 plants had the same level of chlorophyll and carotenoid in comparison with uninfested plants. Our data provide essential information on the effect of D. noxia feeding on chlorophyll and carotenoid concentrations for Betta wheat and its isolines with D. noxia-resistant Dn1 and Dn2 genes.

Animals↗

Chlorophyll degradation in a Chlamydomonas reinhardtii mutant: an accumulation of pyropheophorbide a by anaerobiosis.

Chlorophyll degradation was investigated in cells of a chlorophyll b-less mutant of Chlamydomonas reinhardtii under aerobic and anaerobic conditions. During degradation of chlorophyll under anaerobic conditions, chlorophyll catabolite P535, an open-tetrapyrrole, was not excreted, but pyropheophorbide a was accumulated as the end product with a transient accumulation of chlorophyllide a and pheophorbide a in cells, in contrast to the breakdown under aerobic conditions. It is likely that in the absence of oxygen, degradation of chlorophyll a proceeds to pyropheophorbide a by three consecutive reactions, dephytylation, metal-releasing and demethoxycarbonylation, and then stops due to a limitation of the oxygen that the monooxygenase reaction requires for bilin formation. A novel enzyme catalyzing demethoxycarbonylation of pheophorbide a was partially purified. The enzyme activity increased dependent on the age of cells, and its increase was completely suppressed by cycloheximide. Production of P535 was also dependent on cytoplasmic protein synthesis.

Anaerobiosis↗

Characterization of the Arabidopsis thaliana mutant pcb2 which accumulates divinyl chlorophylls.

We characterized the pcb2 (pale-green and chlorophyll b reduced 2) mutant. We found through electron microscopic observation that chloroplasts of pcb2 mesophyll cells lacked distinctive grana stacks. High-performance liquid chromatography (HPLC) analysis showed that the pcb2 mutant accumulated divinyl chlorophylls, and the relative amount of divinyl chlorophyll b was remarkably less than that of divinyl chlorophyll a. The responsible gene was mapped in an area of 190 kb length at the upper arm of the 5th chromosome, and comparison of DNA sequences revealed a single nucleotide substitution causing a nonsense mutation in At5g18660. Complementation analysis confirmed that the wild-type of this gene suppressed the phenotypes of the mutation. Antisense transformants of the gene also accumulated divinyl chlorophylls. The genes homologous to At5g18660 are conserved in a broad range of species in the plant kingdom, and have similarity to reductases. Our results suggest that the PCB2 product is divinyl protochlorophyllide 8-vinyl reductase.

Arabidopsis↗

Antisense HEMA1 RNA expression inhibits heme and chlorophyll biosynthesis in arabidopsis.

5-aminolevulinic acid (ALA) is a precursor in the biosynthesis of tetrapyrroles including chlorophylls and heme. The formation of ALA involves two enzymatic steps which take place in the chloroplast in plants. The first enzyme, glutamyl-tRNA reductase, and the second enzyme, glutamate-1-semialdehyde-2,1-aminomutase, are encoded by the nuclear HEMA and GSA genes, respectively. To assess the significance of the HEMA gene for chlorophyll and heme synthesis, transgenic Arabidopsis plants that expressed antisense HEMA1 mRNA from the constitutive cauliflower mosaic virus 35S promoter were generated. These plants exhibited varying degrees of chlorophyll deficiency, ranging from patchy yellow to total yellow. Analysis indicated that these plants had decreased levels of chlorophyll, non-covalently bound hemes, and ALA; their levels were proportional to the level of glutamyl-tRNA reductase expression and were inversely related to the levels of antisense HEMA transcripts. Plants that lacked chlorophyll failed to survive under normal growth conditions, indicating that HEMA gene expression is essential for growth.

Aldehyde Oxidoreductases↗

Transbilayer organization of the chlorophyll-proteins of spinach thylakoids.

To investigate the transverse bilayer organization of the chlorophyll-proteins of the three intrinsic chlorophyll-protein complexes, the effects of proteolytic enzymes, and an antibody against the light-harvesting complex were compared using right-side-out and inside-out thylakoid vesicles. The vesicles were isolated by aqueous polymer phase partitioning following the fragmentation of spinach thylakoids by passage through a Yeda press. Both vesicle types were agglutinated by an antiserum specific for the light-harvesting complex, although proteolytic degradation of the complex occurred only in right-side-out vesicles. In addition, there are different antigenic sites for the light-harvesting complex on the inner and outer thylakoid surfaces. Polypeptides of the chlorophyll-alpha-protein complex of photosystem II were degraded by proteases at both membrane surfaces. We concluded that both these chlorophyll-protein complexes are membrane spanning and transversely asymmetric, but that the light-harvesting complex polypeptides accessible at the inner thylakoid surface are more resistant to proteolytic attack. In contrast, the main chlorophyll-containing polypeptide (Mr = 64 500) of photosystem I complex was resistant to proteolytic attack at both the outer and inner thylakoid surfaces.

Chlorophyll↗

Chlorophyll photosensitized electron transfer reactions in lipid vesicles: enhancement in yield of vectorial electron transfer across the bilayer from reduced cytochrome c to oxidized ferredoxin by addition of valinomycin plus potassium ion.

Chlorophyll photosensitized electron transfer across a vesicle bilayer from reduced cytochrome c in the inner compartment to oxidized ferredoxin in the outer compartment, using propylene diquat as a mediator, has been investigated using both steady-state and laser flash photolysis methods. One of the factors limiting the quantum yield is the transmembrane potential, which is formed during sample preparation and is increased by the electron transfer process across the membrane bilayer. This limitation can be diminished by the incorporation of valinomycin into the bilayer in the presence of potassium ion. The overall quantum yield can be approximately doubled (up to a total of 22% based on the chlorophyll triplet which is quenched, and 2.8% based on the absorbed quanta) by valinomycin addition. Another quantum yield limitation arises from the accumulation of oxidized cytochrome c in the inner aqueous compartment, which is formed as a consequence of the transbilayer electron transport process and can quench triplet chlorophyll on the inner side of the vesicle. The chlorophyll cation radical generated in this way can participate in the electron exchange equilibrium between chlorophyll molecules located within the bilayer, and thus inhibit electron flow from inside to outside. This acts to limit the extent of cytochrome c oxidation to less than or equal to 50% of the original amount.

Chlorophyll↗

Effects of light on chloroplast translation in Marchantia polymorpha are similar to those in angiosperms and are not influenced by light-independent chlorophyll synthesis.

Translation of the chloroplast psbA mRNA in angiosperms is activated by photodamage of its gene product, the D1 subunit of photosystem II (PSII), providing nascent D1 for PSII repair. The involvement of chlorophyll in the regulatory mechanism has been suggested due to the regulatory roles of proteins proposed to mediate chlorophyll/D1 transactions and the fact that chlorophyll is synthesized only in the light in angiosperms. We used ribosome profiling and RNA-seq to address whether the effects of light on chloroplast translation are conserved in the liverwort Marchantia (Marchantia polymorpha), which synthesizes chlorophyll in both the dark and the light. As in angiosperms, ribosome occupancy on psbA mRNA decreased rapidly upon shifting plants to the dark and was rapidly restored upon a transfer back to the light, whereas ribosome occupancy on other chloroplast mRNAs changed very little. The results were similar in a Marchantia mutant unable to synthesize chlorophyll in the dark. Those results, in conjunction with pulse-labeling data, suggest that light elicits a plastome-wide activation of translation elongation and a specific increase in psbA translation initiation in Marchantia, as in angiosperms. These findings show that light regulates chloroplast translation similarly in vascular and non-vascular plants, and that constitutive chlorophyll synthesis does not affect light-regulated psbA translation initiation. Additionally, the translational outputs of chloroplast genes are similar in Marchantia and angiosperms but result from differing contributions of mRNA abundance and translational efficiencies. This adds to the evidence that chloroplast mRNA abundance and translational efficiencies co-evolve under selection to maintain protein outputs.

Chloroplasts↗

The effect of cerium (III) on the chlorophyll formation in spinach.

The effect of Ce(3+) on the chlorophyll (chl) of spinach was studied in pot culture experiments. The results showed that Ce(3+) could obviously stimulate the growth of spinach and increase its chlorophyll contents and photosynthetic rate. It could also improve the PSII formation and enhance its electron transport rate of PSII as well. By inductively coupled plasma-mass spectroscopy and atom absorption spectroscopy methods, it was revealed that the rare-earth-element (REE) distribution pattern in the Ce(3+)-treated spinach was leaf > root > shoot in Ce(3+) contents. The spinach leaves easily absorbed REEs. The Ce(3+) contents of chloroplast and chlorophyll of the Ce(3+)-treated spinach were higher than that of any other rare earth and were much higher than that of the control; it was also suggested that Ce(3+) could enter the chloroplast and bind easily to chlorophyll and might replace magnesium to form Ce-chlorophyll. By ultraviolet-visible, Fourier transform infrared, and extended X-ray absorption fine structure (EXAFS) methods, Ce(3+)-coordinated nitrogen of porphyrin rings with eight coordination numbers and average length of the Ce-N bond of 0.251 nm.

Cerium↗

Chlorophyll photobleaching and ethane production in dichlorophenyldimethylurea- (DCMU) or paraquat-treated Euglena gracilis cells.

Light dependent (35 Klux) chlorophyll bleaching in autotrophically grown Euglena gracilis cells at slightly acidic pH (6.5-5.4) is stimulated by the photosystem II blockers DCMU and DBMIB (both 10(-5) M) as well as by the autooxidizable photosystem I electron acceptor, paraquat (10(-3) M). Chlorophyll photobleaching is accompanied by the formation of thiobarbituric acid -- sensitive material ("malondialdehyde") and ethane. Both chlorophyll photobleaching and light dependent ethane formation are partially prevented by higher concentrations (10(-4) M) of the autooxidizable photosystem II electron acceptor DBMIB or by sodium bicarbonate (25 mM). In vitro studies with cell free extracts (homogenates) from E. gracilis suggest that alpha-linolenic acid oxidation by excited (reaction center II) chlorophyll represents the driving force for both ethane formation and chlorophyll bleaching. Ethane formation thus appears to be a sensitive and non-destructive "in vivo" marker for both restricted energy dissipation in photosystem II and, conditions yielding reactive oxygen species at the reducing side of photosystem I.

Chlorophyll↗

Organization and role of the long-wave chlorophylls in the photosystem I of the Cyanobacterium spirulina.

The data on the organization and function of the photosystem I pigment-protein complexes of the cyanobacterium Spirulina and the characteristics of pigment antenna of the photosystem I monomeric and trimeric core complexes are presented and discussed. We proved that the photosystem I complexes in the cyanobacterial membrane pre-exist mainly as trimers, though both types of complexes contribute to the photosynthetic electron transport. In contrast to monomers, the antenna of the photosystem I trimeric complexes of Spirulina contains the extreme long-wave chlorophyll form absorbing at 735 nm and emitting at 760 nm (77 K). The intensity of fluorescence at 760 nm depends strongly on the P700 redox state: it is maximum with the reduced P700 and strongly decreased with the oxidized P700 which is the most efficient quencher of fluorescence at 760 nm. The energy absorbed by the extreme long-wave chlorophyll form is active in the photooxidation of P700 in the trimeric complex. The data obtained indicate that the long-wave form of chlorophyll originates from interaction of the chlorophyll molecules localized on monomeric subunits forming the photosystem I trimer. Kinetic analysis of the P700 photooxidation and light-induced quenching of fluorescence at 760 nm (77 K) allows the suggestion that the excess energy absorbed by the antenna monomeric subunits within the trimer migrates via the extreme long-wave chlorophyll to the P700 cation radical and is quenched, which prevents the photodestruction of the pigment-protein complex.

Chlorophyll↗

[Migration of electron excitation energy in mixed associates of chlorophyll and its derivatives].

The intracomplex migration is investigated of electron excitation energy between molecules which are a part of a mixed associate. Weakly fluorescent and nonfluorescent associates of chlorophyll and its analogues as well as of bacteriochlorophyll and neoxanthine have been used as donors and acceptors. The migration of excitation energy with variation of quantum yield of donor emission from 5-10(-3) to less than or equal to 10(-4) and also with very weak overlapping of luminescence and absorption spectra for the pair D leads to A is experimentally discovered by luminescence excitation spectra of mixed pigment aggregates. A specific disturbing action of Pd and Cu ions on chlorophyll a luminescence with formation of mixed complexes of chlorophyll with Pd-pheophytin a and Cu-pheophytin b is observed. The presence of Pd and Cu ions in the structure of the mixed complexes gives rise to a considerable reduction of chlorophyll apparently due to increasing intersystem crossing probability in the chlorophyll molecule. The probability of intracomplex energy migration in systems under investigation amounts to approximately 10(12) sec-1 according to the obtained estimations. The probable mechanism of energy migration in mixed associates is discussed. The obtained results may be used for elucidation of a possible role of the mixed aggregates and effective intracomplex migration of excitation energy in photosynthetic apparatus of green plants.

Binding Sites↗

QTL and epistatic interaction underlying leaf chlorophyll and H2O2 content variation in rice (Oryza sativa L.).

It is meaningful to study the genetics of the traits associated with photosynthesis such as leaf chlorophyll and H2O2 contents for high yield breeding in rice. A mapping population of 98 BC1F9 lines (backcross Inbred Lines, BIL), derived from a backcross of Nipponbare (japonica)/Kasalath(indica)//Nipponbare by the single-seed-descent methods, was employed to map quantitative trait loci (QTL) underlying such rice traits as leaf chlorophyll and H2O2 contents. Five and two QTLs were detected for leaf Chlorophyll and H2O2 content variation, respectively. Among the QTLs detected, q-Chl1 on chromosome 1 accounted for 22% variation for leaf chlorophyll content variation. The alleles from Nipponbare increased stature at the locus; On the other hand, q-H2O2(1) which associated with H2O2 content was also located on the same region as q-Chl1. The alleles from Nipponbare decreased stature at this locus. The 2 traits have the 2 QTL sharing the same chromosome locations, which was located between C86 and C813 on chromosome 1. Epistasis analysis showed QTL with interaction distributed on chromosome 2, 6, 11 and 12. However, no QTL interacted with q-Chl1 or q-H2O2(1) was detected. It can be deduced that alleles from Nipponbare at the region between C86 and C813 on chromosome 1 increases leaf chlorophyll content and decreases H2O2 content at the mean time with significant additive effect but little epistasis.

Chlorophyll↗

[Chlorophyll a content and primary productivity of phytoplankton in Meiliang Bay of Taihu Lake].

Based on the investigations on primary productivity of phytoplankton and related factors at four stations in Meiliang Bay of Taihu Lake from May 1998 to August 1999, this paper studied the distribution of phytoplankton chlorophyll a, its primary productivity, and their relationships with environmental factors. The results showed that there were obvious temporal and spatial differences in chlorophyll a and primary productivity. The high values of chlorophyll a content and primary productivity were recorded in spring and summer. Generally, chlorophyll a content and primary productivity decreased from inner to outer in the bay. The vertical profile of primary productivity showed that the maximum of primary productivity was recorded at 20-50 cm water depth in spring, summer and autumn, while there was no remarkable difference in winter. The primary productivity exponentially increased with water temperature from 10 to 30 degrees C. There was a significant positive correlation between chlorophyll a content and primary productivity. The correlation coefficients between nutrients and primary productivity varied greatly, which presented that nutrient was not the limiting factor for phytoplankton growth. The analysis of P-I curves based on different photosynthesis-irradiance models showed that obvious light inhibition was occurred at surface water in spring and summer.

Biomass↗

[Application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study].

The changes in PSII performance of the photosynthetic apparatus caused by environmental stress or senescence have been explored widely by applying the chlorophyll fluorescence technique, mainly by using the modulated fluorometer. In recent years, an alternative approach via analyzing chlorophyll a fluorescence transient using the continuous excitation fluorometer has been developed, which offers more information that cannot be obtained by the modulated fluorometry. The JIP-test based on the theory of energy fluxes in biomembranes has been widely used to analyze the chlorophyll a fluorescence kinetics transient helping researchers to have a deeper insight into the primary photochemical reaction in the photosynthetic apparatus. In this paper, the chlorophyll a fluorescence transient and its significance, the parameters involved and their significance in the JIP-test, and the application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study are introduced combining some results of the author's.

Chlorophyll↗

Role of L-alanine: 4,5-dioxovalerate transaminase in chlorophyll synthesis in Bajra (Pennisetum typhoideum) seedlings.

L-alanine:4,5-dioxovalerate transaminase activity and chlorophyll levels were estimated in lead and mercury treated Bajra seedlings. The enzyme activity increased with age upto 2nd day of germination and decreased on consequent days, where as the chlorophyll content increased with age upto 4th day and remained constant on day 5. Both the metals have no effect on L-alanine: 4,5-dioxovalerate transaminase activity but reduced chlorophyll levels. In vitro incubation of the enzyme with metal solutions showed that the enzyme activity was inhibited by mercury, while lead had no effect. Studies on sub-cellular localization of the L-alanine:4,5-dioxovalerate transaminase showed that it is present in all fractions. The non-correlation between L-alanine: 4,5-dioxovalerate transaminase activity and chlorophyll synthesis is evident from different activity profiles with age and response to heavy metal treatment in the seedlings. Hence, our results suggest the non-involvement of L-alanine:4,5-dioxovalerate transaminase in chlorophyll synthesis in bajra seedlings.

Aminolevulinic Acid↗

Two partially homologous adjacent light-inducible maize chloroplast genes encoding polypeptides of the P700 chlorophyll a-protein complex of photosystem I.

The maize chloroplast chromosome contains two light-inducible genes, ps1A1 and ps1A2, that code for 45% homologous polypeptides of 83.2 and 82.5 kDa designated A1 and A2, respectively. Two types of immunochemical evidence show that the upstream gene, ps1A1, codes for a P700 chlorophyll a-protein at the reaction center of photosystem I of the photosynthetic apparatus. Antibodies against a synthetic peptide with a sequence deduced from the DNA sequence of an unconserved segment of A1 react with polypeptides of P700 chlorophyll-protein (CPI) complexes of maize and pea photosystem I; antibodies prepared against barley CPI immunoprecipitate products of in vitro transcription and translation directed by cloned chloroplast DNA containing this gene. The extensive homology between maize polypeptides A1 and A2 suggests that both may be components of CPI, although CPI has been generally considered to be comprised of two molecules of a single protein of only 66 to 70 kDa. The hexapeptide Asp-Pro-Thr-Thr-Arg-Arg in A2 is also present in another chlorophyll protein, the P680 chlorophyll a-protein of photosystem II, and is partially duplicated in A1. The number and locations of histidyl residues, which have been suggested to serve in chlorophyll binding, are highly conserved between A1 and A2.

Amino Acid Sequence↗

Absorption of chlorophyll phytol in normal man and in patients with Refsum's disease.

This study was made to determine the extent of absorption of chlorophyll phytol from the intestine of man, and the importance of chlorophyll as a source of the phytanic acid that accumulates in Refsum's disease. Uniformly (14)C-labeled pheophytin a (the Mg-free derivative of chlorophyll a) was fed to normal human subjects and to patients with Refsum's disease. Feces were collected and analyzed. In all subjects, 90-95% of the administered radioactivity was recovered in the feces, still largely in the form of pheophytin a. The phytol radioactivity recovered in the feces averaged about 95% of that in the administered material, which indicates that there had been little absorption of the phytol moiety. Similarly, after 250 g of cooked spinach had been fed to a normal subject, almost the entire phytol content was found in the feces. Less than 5% of the ingested spinach phytol was accounted for in the thoracic duct lymph of another subject. It was concluded that not more than about 5% of the ingested chlorophyll phytol is absorbed by man, whether normal or afflicted with Refsum's disease. On this basis we conclude that the major portion of the phytanic acid that accumulates in Refsum's disease could not be derived from dietary chlorophyll.

Adolescent↗

[Spectral forms of the chlorophyll of mutants of Chlamydomonas with inactive photosystems].

Investigations of non-photosinthesizing mutants of Chlamydomonas reinhardii with damaged activity of both or one of the photosystems have shown that the chlorophyll a form with the absorption maximum at 685 nm and low temperature fluorescence band at 696 nm is a part of the pigment-protein complex of PS-2. Chlorophyll a forms with absorption maxima at 689, 698 and 703 nm belong to the pigment-protein complex of PS-1. They seem to be responsible for the long-wave band of chlorophyll fluorescence with the maximum at 707--718 nm. Chlorophyll a forms with absorption at 661, 667 and 678 nm and a short-wave fluorescence band at 685--690 nm rank among the pigments of the light-converging complex, as well as chlorophyll b with the absorption maximum at 644 and 649 nm.

Chemical Phenomena↗