Chlorophyllide a Oxygenase mRNA and Protein Levels Correlate with the Chlorophyll a/b Ratio in Arabidopsis thaliana (original) (raw)

Light Intensity-Dependent Modulation of Chlorophyll b Biosynthesis and Photosynthesis by Overexpression of Chlorophyllide a Oxygenase in Tobacco

Chlorophyll b is synthesized by the oxidation of a methyl group on the B ring of a tetrapyrrole molecule to a formyl group by chlorophyllide a oxygenase (CAO). The full-length CAO from Arabidopsis (Arabidopsis thaliana) was overexpressed in tobacco (Nicotiana tabacum) that grows well at light intensities much higher than those tolerated by Arabidopsis. This resulted in an increased synthesis of glutamate semialdehyde, 5-aminolevulinic acid, magnesium-porphyrins, and chlorophylls. Overexpression of CAO resulted in increased chlorophyll b synthesis and a decreased chlorophyll a/b ratio in low light grown as well as high light-grown tobacco plants; this effect, however, was more pronounced in high light. The increased potential of the protochlorophyllide oxidoreductase activity and chlorophyll biosynthesis compensated for the usual loss of chlorophylls in high light. Increased chlorophyll b synthesis in CAO-overexpressed plants was accompanied not only by an increased abundance of light-harvesting chlorophyll proteins but also of other proteins of the electron transport chain, which led to an increase in the capture of light as well as enhanced (40%–80%) electron transport rates of photosystems I and II at both limiting and saturating light intensities. Although the quantum yield of carbon dioxide fixation remained unchanged, the lightsaturated photosynthetic carbon assimilation, starch content, and dry matter accumulation increased in CAO-overexpressed plants grown in both low- and high-light regimes. These results demonstrate that controlled up-regulation of chlorophyll b biosynthesis comodulates the expression of several thylakoid membrane proteins that increase both the antenna size and the electron transport rates and enhance carbon dioxide assimilation, starch content, and dry matter accumulation.

Expression of Enzymes Involved in Chlorophyll Catabolism in Arabidopsis Is Light Controlled

PLANT PHYSIOLOGY, 2011

We found that the levels of mRNA of two enzymes involved in chlorophyll catabolism in Arabidopsis (Arabidopsis thaliana), products of two chlorophyllase genes, AtCLH1 and AtCLH2, dramatically increase (by almost 100-and 10-fold, respectively) upon illumination with white light. The measurements of photosystem II quantum efficiency in 3-(3,4-dichlorophenyl)-1,1dimethylurea-inhibited leaves show that their expression is not related to photosynthesis but mediated by photoreceptors. To identify the photoreceptors involved, we used various light treatments and Arabidopsis photoreceptor mutants (cry1, cry2, cry1cry2, phot1, phot2, phot1phot2, phyA phyB, phyAphyB). In wild-type Columbia, the amount of transcripts of both genes increase after white-light irradiation but their expression profile and the extent of regulation differ considerably. Blue and red light is active in the case of AtCLH1, whereas only blue light raises the AtCLH2 mRNA level. The fundamental difference is the extent of up-regulation, higher by one order of magnitude in AtCLH1. Both blue and red light is active in the induction of AtCLH1 expression in all mutants, pointing to a complex control network and redundancy between photoreceptors. The bluespecific up-regulation of the AtCLH2 transcript is mediated by cryptochromes and modulated by phototropin1 and phytochromes. Individually darkened leaves were used to test the effects of senescence on the expression of AtCLH1 and AtCLH2. The expression profile of AtCLH1 remains similar to that found in nonsenescing leaves up to 5 d after darkening. In contrast, the light induction of AtCLH2 mRNA declines during dark treatment. These results demonstrate that the expression of enzymes involved in chlorophyll catabolism is light controlled.

Light-dependent regulation of chlorophyll b biosynthesis in chlorophyllide a oxygenase overexpressing tobacco plants

Biochemical and Biophysical Research Communications, 2005

Chlorophyllide a oxygenase (CAO) that converts chlorophyllide a to chlorophyllide b was overexpressed in tobacco to increase chlorophyll (Chl) b biosynthesis and alter the Chl a/b ratio. Transgenic plants along with their wild-type cultivars were grown in low and high light intensities. In low light there was 20% increase in chlorophyll b contents in transgenic plants, which resulted in 16% reduction in the Chl a/b ratio. In high light, total Chl contents were 31% higher in transgenic plants than those of wild type. The increase in Chl a was 19% and that of Chl b was 72% leading to 31% decline of Chl a/b ratio. The increase in Chl b contents was accompanied by enhanced CAO expression that was highly pronounced in low light. As compared to low light, in high light Lhcb1 and Chl a/b transcripts abundance was significantly increased in transgenic plants suggesting a close relationship between Chl b synthesis and cab gene expression. However, there was a small increase in expression of LHCII proteins, which did not correspond to 72% increase in Chl b content in transgenic line, implying that LHCPII has the ability to bind more Chl b molecules.

Light Intensity-Dependent Modulation of Chlorophyll b Biosynthesis and Photosynthesis by Overexpression of Chlorophyllide a Oxygenase (CAO) in Tobacco

Plant physiology, 2012

Chlorophyll b is synthesized by the oxidation of a methyl group on the B ring of a ttrapyrrole molecule to a formyl group by chlorophyllide a oxygenase (CAO). The full length CAO, from Arabidopsis thaliana was overexpressed in tobacco (Nicotiana tabacum) that grows well at light intensities much higher than those tolerated by Arabidopsis. This resulted in an increased synthesis of glutamate semialdehyde, 5-aminolevulinicacid, Mg-porphyrins and chlorophylls. Overexpression of CAO resulted in increased chlorophyll b synthesis and a decreased chlorophyll a/b ratio in low-light-grown as well as in high-light-grown tobacco plants; this effect was, however, more pronounced in high-light. The increased potential of the protochlorophyllide oxidoreductase activity and chlorophyll biosynthesis compensated for the usual loss of chlorophylls in high light. Increased chlorophyll b synthesis in CAO-overexpressed plants was accompanied not only by an increased abundance of light-harvesting chlorop...

Deregulated Chlorophyll b Synthesis Reduces the Energy Transfer Rate Between Photosynthetic Pigments and Induces Photodamage in Arabidopsis thaliana

Plant and Cell Physiology, 2010

Chl b is one of the major light-harvesting pigments in land plants. The synthesis of Chl b is strictly regulated in response to light conditions in order to control the antenna size of photosystems. Regulation of Chl b also affects its distribution as it occurs preferentially in the peripheral antenna complexes. However, it has not been experimentally shown how plants respond to environmental conditions when they accumulate excess Chl b. Previously, we produced an Arabidopsis transgenic plant (referred to as the BC plant) in which Chl b biosynthesis was enhanced. In this study, we analyzed the photosynthetic properties and genome-wide gene expression in this plant under high light conditions in order to understand the effects of deregulated Chl b biosynthesis. The energy transfer rates between Chl a molecules in PSII decreased and H 2 O 2 accumulated extensively in the BC plant. Microarray analysis revealed that a group of genes involved in anthocyanin biosynthesis was downregulated and that another group of genes, reported to be sensitive to H 2 O 2 , was up-regulated in the BC plant. We also found that anthocyanin levels were low, which was consistent with the results of the microarray analysis. These results indicate that deregulation of Chl b caused severe photodamage and altered gene expression profi les under strong illumination. The importance of the regulation of Chl b synthesis is discussed in relation to the correct localization of Chl b and gene expression.

Photocontrol of the Expression of Genes Encoding Chlorophyll a/b Binding Proteins and Small Subunit of Ribulose-1,5-Bisphosphate Carboxylase in Etiolated Seedlings of Lycopersicon esculentum (L.) and Nicotiana tabacum (L.)

Plant Physiology, 1990

Phytochrome and the blue ultraviolet-A photoreceptor control light-induced expression of genes encoding the chlorophyll a/b binding protein of photosystem 11 and photosystem I and the genes for the small subunit of the ribulose-1,5-bisphosphate carboxylase in etiolated seedlings of Lycopersicon esculentum (tomato) and Nicotiana tabacum (tobacco). A 'high irradiance response' also controls the induction of these genes. Gernes encoding photosystem 11-and I-associated chlorophyll a/b binding proteins both exhibit a transient rapid increase in expression in response to light pulse or to continuous irradiation. In contrast, genes encoding the small subunit exhibit a continuous increase in expression in response to light. These distinct expression characteristics are shown to reflect differences at the level of transcription. Higher plants have several photoreceptors which detect light quality and intensity. The major photoreceptors are phytochrome (26), which control induction and 'high irradiance responses' (HIR2) (10), a blue/UV-A photoreceptor (27), and a UV-B photoreceptor (34). Studies of expression of nuclear genes encoding CAB, SSU, and CHS proteins revealed that photoregulation of gene expression in higher plants occurs at transcriptional and posttranscriptional levels (33). The dependence ofgene expression on light quality and intensity varies for different genes and different species. For instance, in parsley cell suspension cultures excitation of the UV-B photoreceptor is essential for maximal expression of CHS, while the excitation of the blue/

Compensation Mechanism of the Photosynthetic Apparatus in Arabidopsis thaliana ch1 Mutants

International Journal of Molecular Sciences

The origin of chlorophyll b deficiency is a mutation (ch1) in chlorophyllide a oxygenase (CAO), the enzyme responsible for Chl b synthesis. Regulation of Chl b synthesis is essential for understanding the mechanism of plant acclimation to various conditions. Therefore, the main aim of this study was to find the strategy in plants for compensation of low chlorophyll content by characterizing and comparing the performance and spectral properties of the photosynthetic apparatus related to the lipid and protein composition in four selected Arabidopsis ch1 mutants and two Arabidopsis ecotypes. Mutation in different loci of the CAO gene, viz., NW41, ch1.1, ch1.2 and ch1.3, manifested itself in a distinct chlorina phenotype, pigment and photosynthetic protein composition. Changes in the CAO mRNA levels and chlorophyllide a (Chlide a) content in ecotypes and ch1 mutants indicated their significant role in the adjustment mechanism of the photosynthetic apparatus to low-light conditions. Expo...

Modulation of Chlorophyll b Biosynthesis and Photosynthesis by Overexpression of Chlorophyllide a Oxygenase (CAO) in Tobacco

Advanced Topics in Science and Technology in China, 2013

Chlorophyll (Chl) b is synthesized by oxidation of a methyl group on the B ring of the porphyrin molecule to a formyl group by chlorophyllide (Chlide) a oxygenase (CAO). The overexpression of Arabidopsis thaliana full length CAO (AtCAO) in tobacco (Nicotiana tabacum) resulted in an increased Chl synthesis and a decreased Chl a/b ratio in low-light-grown (LL) as well as in highlight grown (HL) tobacco plants, where the effect was more pronounced. In HL-plants, increased [Chl b] resulted in efficient capture of solar energy and enhanced (40%-80%) electron transport rates at both limiting and saturating light intensities.

Light quantity controls leaf-cell and chloroplast development in Arabidopsis thaliana wild type and blue-light-perception mutants

Planta, 2000

Plants acclimate to changes in light quantity by altering leaf-cell development and the accumulation of chloroplast components, such that light absorption is favoured under limiting illumination, and light utilisation under non-limiting conditions. Previous evidence suggests an involvement of a high-light photosynthetic redox signal in the down-regulation of the accumulation of the light-harvesting complexes of photosystem II (Lhcb) and the expression of the Lhcb genes, and of a blue-light signal in the control of leaf development and in the increase in photosynthetic capacity, as aected by the accumulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). We examined the internal anatomy of leaves, the ultrastructure of chloroplasts and accumulation of light-harvesting complexes and Rubisco in wild-type Arabidopsis thaliana (L.) Heynh. and in mutants in each of the three known blue-light photoreceptors, cryptochrome 1, cryptochrome 2 and phototropin, as well as a mutant in both cryptochromes. Our results indicate an extensive capacity of the Arabidopsis mesophyll cells to adapt to high light¯uence rate with an increase in palisade elongation. Under high light, chloroplasts showed increased starch accumulation and reductions in the amount of granal thylakoids per chloroplast, in the proportion of chlorophyll b relative to chlorophyll a, and in the accumulation of the major Lhcb polypeptides. The responses were similar for all four mutants, with respect to their wild types. The results are consistent with either a complete redundancy in function between cryptochromes and phototropin, or their absence of involvement in the light-quantity responses tested. We observed minimal eects of light quantity on Rubisco accumulation over the range of¯uence rates used, and conclude that elongation of palisade mesophyll cells and accumulation of Rubisco are controlled separately. This indicates that light acclimation must be the result of a number of individual elementary responses. Quantitative dierences in the acclimatory responses were observed between the Landsberg erecta and Columbia wild-type ecotypes used.

The AtCAO gene, encoding chlorophyll a oxygenase, is required for chlorophyll b synthesis in Arabidopsis thaliana

Proceedings of the …, 1999

Chlorophyll b is synthesized from chlorophyll a and is found in the light-harvesting complexes of prochlorophytes, green algae, and both nonvascular and vascular plants. We have used conserved motifs from the chlorophyll a oxygenase (CAO) gene from Chlamydomonas reinhardtii to isolate a homologue from Arabidopsis thaliana. This gene, AtCAO, is mutated in both leaky and null chlorina1 alleles, and DNA sequence changes cosegregate with the mutant phenotype. AtCAO mRNA levels are higher in three different mutants that have reduced levels of chlorophyll b, suggesting that plants that do not have sufficient chlorophyll b up-regulate AtCAO gene expression. Additionally, AtCAO mRNA levels decrease in plants that are grown under dimlight conditions. We have also found that the six major Lhcb proteins do not accumulate in the null ch1-3 allele.