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ABSTRACT: Xanthophylls are oxygenated carotenoids playing an essential role as structural components of the photosynthetic apparatus. Xanthophylls contribute to the assembly and stability of light-harvesting complex, to light absorbance and to photoprotection. The first step in xanthophyll biosynthesis from α- and β-carotene is the hydroxylation of ε- and β-rings, performed by both non-heme iron oxygenases (CHY1, CHY2) and P450 cytochromes (LUT1/CYP97C1, LUT5/CYP97A3). The Arabidopsis triple chy1chy2lut5 mutant is almost completely depleted in β-xanthophylls.
Here we report on the quadruple chy1chy2lut2lut5 mutant, additionally carrying the lut2 mutation (affecting lycopene ε-cyclase). This genotype lacks lutein and yet it shows a compensatory increase in β-xanthophylls with respect to chy1chy2lut5 mutant. Mutant plants show an even stronger photosensitivity than chy1chy2lut5, a complete lack of qE, the rapidly reversible component of non-photochemical quenching, and a peculiar organization of the pigment binding complexes into thylakoids. Biochemical analysis reveals that the chy1chy2lut2lut5 mutant is depleted in Lhcb subunits and is specifically affected in Photosystem I function, showing a deficiency in PSI-LHCI supercomplexes. Moreover, by analyzing a series of single, double, triple and quadruple Arabidopsis mutants in xanthophyll biosynthesis, we show a hitherto undescribed correlation between xanthophyll levels and the PSI-PSII ratio. The decrease in the xanthophyll/carotenoid ratio causes a proportional decrease in the LHCII and PSI core levels with respect to PSII.
The physiological and biochemical phenotype of the chy1chy2lut2lut5 mutant shows that (i) LUT1/CYP97C1 protein reveals a major β-carotene hydroxylase activity in vivo when depleted in its preferred substrate α-carotene; (ii) xanthophylls are needed for normal level of Photosystem I and LHCII accumulation.
BMC Plant Biology 04/2012; 12:50. · 3.45 Impact Factor
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ABSTRACT: Xanthophylls (oxygenated carotenoids) are essential components of the plant photosynthetic apparatus, where they act in photosystem assembly, light harvesting, and photoprotection. Nevertheless, the specific function of individual xanthophyll species awaits complete elucidation. In this work, we analyze the photosynthetic phenotypes of two newly isolated Arabidopsis mutants in carotenoid biosynthesis containing exclusively alpha-branch (chy1chy2lut5) or beta-branch (chy1chy2lut2) xanthophylls. Both mutants show complete lack of qE, the rapidly reversible component of nonphotochemical quenching, and high levels of photoinhibition and lipid peroxidation under photooxidative stress. Both mutants are much more photosensitive than npq1lut2, which contains high levels of viola- and neoxanthin and a higher stoichiometry of light-harvesting proteins with respect to photosystem II core complexes, suggesting that the content in light-harvesting complexes plays an important role in photoprotection. In addition, chy1chy2lut5, which has lutein as the only xanthophyll, shows unprecedented photosensitivity even in low light conditions, reduced electron transport rate, enhanced photobleaching of isolated LHCII complexes, and a selective loss of CP26 with respect to chy1chy2lut2, highlighting a specific role of beta-branch xanthophylls in photoprotection and in qE mechanism. The stronger photosystem II photoinhibition of both mutants correlates with the higher rate of singlet oxygen production from thylakoids and isolated light-harvesting complexes, whereas carotenoid composition of photosystem II core complex was not influential. In depth analysis of the mutant phenotypes suggests that alpha-branch (lutein) and beta-branch (zeaxanthin, violaxanthin, and neoxanthin) xanthophylls have distinct and complementary roles in antenna protein assembly and in the mechanisms of photoprotection.
Journal of Biological Chemistry 12/2007; 282(48):35056-68. · 4.77 Impact Factor
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ABSTRACT: The first dedicated step in plant xanthophyll biosynthesis is carotenoid hydroxylation. In Arabidopsis thaliana, this reaction is performed by both heme (LUT1 and LUT5) and non-heme (CHY1 and CHY2) hydroxylases. No mutant completely abolishing alpha- or beta-carotene hydroxylation has been described to date. We constructed double and triple mutant combinations in CHY1, CHY2, LUT1, LUT5 and LUT2 (lycopene epsilon-cyclase). In chy1chy2lut2, 80% of leaf carotenoids is represented by beta-carotene. In chy1chy2lut5, beta-carotene hydroxylation is completely abolished, while hydroxylation of the beta-ring of alpha-carotene is still observed. The data are consistent with a role of LUT5 in beta-ring hydroxylation, and with the existence of an additional hydroxylase, acting on the beta-ring of alpha-, but not beta-carotene.
FEBS Letters 09/2006; 580(19):4718-22. · 3.54 Impact Factor
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Claudia Dalmastri, Alessia Fiore,
Chiara Alisi,
Annamaria Bevivino,
Silvia Tabacchioni,
Giovanni Giuliano,
Anna Rosa Sprocati,
Lia Segre,
Eshwar Mahenthiralingam,
Luigi Chiarini,
Peter Vandamme
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ABSTRACT: The Burkholderia cepacia‘complex’ (Bcc) presently comprises nine species and genomovars. In order to acquire a better comprehension of the species and genomovar distribution and of the genetic diversity among environmental Bcc bacteria, a natural population of 60 bacterial isolates recovered from the rhizosphere of maize and belonging to the Bcc has been characterised to assess the exact taxonomic position, the genetic polymorphism and the metabolic profiles of isolates. The identification of the different species and genomovars was accomplished by a combination of techniques including sodium dodecyl sulfate–polyacrylamide gel electrophoresis of whole-cell proteins and recA-based restriction fragment length polymorphism analyses. The genetic diversity among Bcc isolates was analysed by means of the random amplified polymorphic DNA and amplified fragment length polymorphism techniques; the analysis of molecular variance method was applied to estimate the genetic differences among the various species and genomovars identified within the bacterial population. Metabolic profiles based on carbon source utilisation were obtained by means of the Biolog GN assay and analysed by means of cluster analysis. Forty-four strains were identified as B. ambifaria, 11 as B. cenocepacia recA lineage III-B, four as B. pyrrocinia, and one as B. cepacia genomovar I. Marked genetic differences were observed between B. cenocepacia and B. ambifaria, whereas limited differences were found between B. pyrrocinia and B. ambifaria and between B. pyrrocinia and B. cenocepacia. No significant differences (P>0.05) were observed between the mean genetic distances of isolates belonging to B. cenocepacia, B. ambifaria, and B. pyrrocinia. Phenotypic analyses revealed that all isolates tested were able to utilise more than 75% of substrates. The highest variability in the number of utilised substrates was found among B. cenocepacia isolates, whereas the lowest was found among B. ambifaria isolates. Cluster analysis of metabolic profiles revealed pronounced differences between B. cenocepacia and B. ambifaria; in contrast, B. pyrrocinia could not be clearly separated either from B. cenocepacia or from B. ambifaria.
FEMS Microbiology Ecology 01/2006; 46(2):179 - 187. · 3.41 Impact Factor
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ABSTRACT: Cryptochromes are blue light photoreceptors found in plants, bacteria, and animals. In Arabidopsis, cryptochrome 2 (cry2) is involved primarily in the control of flowering time and in photomorphogenesis under low-fluence light. No data on the function of cry2 are available in plants, apart from Arabidopsis (Arabidopsis thaliana). Expression of the tomato (Solanum lycopersicum) CRY2 gene was altered through a combination of transgenic overexpression and virus-induced gene silencing. Tomato CRY2 overexpressors show phenotypes similar to but distinct from their Arabidopsis counterparts (hypocotyl and internode shortening under both low- and high-fluence blue light), but also several novel ones, including a high-pigment phenotype, resulting in overproduction of anthocyanins and chlorophyll in leaves and of flavonoids and lycopene in fruits. The accumulation of lycopene in fruits is accompanied by the decreased expression of lycopene beta-cyclase genes. CRY2 overexpression causes an unexpected delay in flowering, observed under both short- and long-day conditions, and an increased outgrowth of axillary branches. Virus-induced gene silencing of CRY2 results in a reversion of leaf anthocyanin accumulation, of internode shortening, and of late flowering in CRY2-overexpressing plants, whereas in wild-type plants it causes a minor internode elongation.
Plant physiology 02/2005; 137(1):199-208. · 6.53 Impact Factor
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ABSTRACT: In order to acquire a better understanding of the effects of the different delivery modes of bacterial inoculants on plant growth and on the community structure of rhizosphere bacterial populations, Burkholderia ambifaria MCI 7 (formerly B. cepacia MCI 7) was inoculated into the rhizosphere of maize plants by either seed adhesion or incorporation into soil. Plant growth was evaluated at different inoculum concentrations. The community structure of rhizosphere bacterial populations was evaluated by analysing the restriction patterns of the DNA coding for 16S rRNA amplified by polymerase chain reaction (PCR) (ARDRA) of 745 bacterial isolates. A number of diversity indices (richness, Shannon diversity, evenness and mean genetic distance) were calculated for each bacterial population isolated from control and treated plants according to the concept of the r/K strategy. Moreover, the analysis of molecular variance (AMOVA) method was applied to estimate the genetic differences among the various bacterial populations. Our results showed that the method of application can be an essential element in determining the effects of the inoculant on plant growth. In fact, when applied as a maize seed treatment, B. ambifaria MCI 7 promoted plant growth significantly; on the contrary, when incorporated into soil, the same strain reduced plant growth markedly. As far as the bacterial community structure is concerned, B. ambifaria MCI 7 affected the indigenous microflora of treated plants according to the application method: seed treatment brought about an abrupt decrease in bacterial diversity, whereas incorporation into soil increased bacterial diversity. Moreover, changes in bacterial diversity were limited to r-strategist bacteria. In conclusion, B. ambifaria MCI 7 can act as both a plant growth-promoting rhizobacterium and a deleterious rhizobacterium depending on the inoculation method.
Environmental Microbiology 05/2002; 4(4):238-45. · 5.84 Impact Factor
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ABSTRACT: The first dedicated step in plant xanthophyll biosynthesis is carotenoid hydroxylation. In Arabidopsis thaliana, this reaction is performed by both heme (LUT1 and LUT5) and non-heme (CHY1 and CHY2) hydroxylases. No mutant completely abolishing α- or β-carotene hydroxylation has been described to date. We constructed double and triple mutant combinations in CHY1, CHY2, LUT1, LUT5 and LUT2 (lycopene epsilon-cyclase). In chy1chy2lut2, 80% of leaf carotenoids is represented by β-carotene. In chy1chy2lut5, β-carotene hydroxylation is completely abolished, while hydroxylation of the β-ring of α-carotene is still observed. The data are consistent with a role of LUT5 in β-ring hydroxylation, and with the existence of an additional hydroxylase, acting on the β-ring of α-, but not β-carotene.
FEBS Letters.
