scholarly journals Genetic manipulation of carotenoid biosynthesis and photoprotection

2000 ◽  
Vol 355 (1402) ◽  
pp. 1395-1403 ◽  
Author(s):  
Barry J. Pogson ◽  
Heather M. Rissler
Keyword(s):  
Xanthophyll Cycle ◽  
Genetic Manipulation ◽  
Carotenoid Biosynthesis ◽  
Photochemical Quenching ◽  
Light Harvesting Complex ◽  
Photooxidative Damage ◽  
Lutein Content ◽  
Genetically Manipulated ◽  
Non Photochemical Quenching ◽  
Β Carotene

There are multiple complementary and redundant mechanisms to provide protection against photooxidative damage, including non–photochemical quenching (NPQ). NPQ dissipates excess excitation energy as heat by using xanthophylls in combination with changes to the light–harvesting complex (LHC) antenna. The xanthophylls are oxygenated carotenoids that in addition to contributing to NPQ can quench singlet or triplet chlorophyll and are necessary for the assembly and stability of the antenna. We have genetically manipulated the expression of the ε–cyclase and β–carotene hydroxylase carotenoid biosynthetic enzymes in Arabidopsis thaliana . The ε–cyclase overexpression confirmed that lut2 (lutein deficient) is a mutation in the ε–cyclase gene and demonstrated that lutein content can be altered at the level of mRNA abundance with levels ranging from 0 to 180% of wild–type. Also, it is clear that lutein affects the induction and extent of NPQ. The deleterious effects of lutein deficiency on NPQ in Arabidopsis and Chlamydomonas are additive, no matter what the genetic background, whether npq1 (zeaxanthin deficient), aba1 or antisense β–hydroxylase (xanthophyll cycle pool decreased). Additionally, increasing lutein content causes a marginal, but significant, increase in the rate of induction of NPQ despite a reduction in the xanthophyll cycle pool size.

Non-photochemical quenching of chlorophyll a fluorescence: early history and characterization of two xanthophyll-cycle mutants of Chlamydomonas reinhardtii

2002 ◽  
Vol 29 (10) ◽  
pp. 1141 ◽  
Author(s):  
Govindjee ◽  
Manfredo J. Seufferheld
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Oxygen Evolution ◽  
Xanthophyll Cycle ◽  
Photochemical Quenching ◽  
Chl A ◽  
Fluorescence Transient ◽  
Chl A Fluorescence ◽  
Non Photochemical Quenching

This paper deals first with the early, although incomplete, history of photoinhibition, of 'non-QA-related chlorophyll (Chl) a fluorescence changes', and the xanthophyll cycle that preceded the discovery of the correlation between non-photochemical quenching of Chl a fluorescence (NPQ) and conversion of violaxanthin to zeaxanthin. It includes the crucial observation that the fluorescence intensity quenching, when plants are exposed to excess light, is indeed due to a change in the quantum yield of fluorescence. The history ends with a novel turn in the direction of research — isolation and characterization of NPQ xanthophyll-cycle mutants of Chlamydomonas reinhardtii Dangeard and Arabidopsis thaliana (L.) Heynh., blocked in conversion of violaxanthin to zeaxanthin, and zeaxanthin to violaxanthin, respectively. In the second part of the paper, we extend the characterization of two of these mutants (npq1, which accumulates violaxanthin, and npq2, which accumulates zeaxanthin) through parallel measurements on growth, and several assays of PSII function: oxygen evolution, Chl a fluorescence transient (the Kautsky effect), the two-electron gate function of PSII, the back reactions around PSII, and measurements of NPQ by pulse-amplitude modulation (PAM 2000) fluorimeter. We show that, in the npq2 mutant, Chl a fluorescence is quenched both in the absence and presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). However, no differences are observed in functioning of the electron-acceptor side of PSII — both the two-electron gate and the back reactions are unchanged. In addition, the role of protons in fluorescence quenching during the 'P-to-S' fluorescence transient was confirmed by the effect of nigericin in decreasing this quenching effect. Also, the absence of zeaxanthin in the npq1 mutant leads to reduced oxygen evolution at high light intensity, suggesting another protective role of this carotenoid. The available data not only support the current model of NPQ that includes roles for both pH and the xanthophylls, but also are consistent with additional protective roles of zeaxanthin. However, this paper emphasizes that we still lack sufficient understanding of the different parts of NPQ, and that the precise mechanisms of photoprotection in the alga Chlamydomonas may not be the same as those in higher plants.

scholarly journals Identification of parallel pH- and zeaxanthin-dependent quenching of excess energy in LHCSR3 in Chlamydomonas reinhardtii

2020 ◽  
Author(s):  
Julianne M. Troiano ◽  
Federico Perozeni ◽  
Raymundo Moya ◽  
Luca Zuliani ◽  
Kwangryul Baek ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Light Harvesting ◽  
Complex Stress ◽  
Excess Energy ◽  
Photochemical Quenching ◽  
Light Harvesting Complexes ◽  
Light Harvesting Complex ◽  
Non Photochemical Quenching

AbstractUnder high light conditions, oxygenic photosynthetic organisms avoid photodamage by thermally dissipating excess absorbed energy, which is called non-photochemical quenching (NPQ). In green algae, a chlorophyll and carotenoid-binding protein, light-harvesting complex stress-related (LHCSR3), detects excess energy via pH and serves as a quenching site. However, the mechanisms by which LHCSR3 functions have not been determined. Using a combined in vivo and in vitro approach, we identify two parallel yet distinct quenching processes, individually controlled by pH and carotenoid composition, and their likely molecular origin within LHCSR3 from Chlamydomonas reinhardtii. The pH-controlled quenching is removed within a mutant LHCSR3 that lacks the protonable residues responsible for sensing pH. Constitutive quenching in zeaxanthin-enriched systems demonstrates zeaxanthin-controlled quenching, which may be shared with other light-harvesting complexes. We show that both quenching processes prevent the formation of damaging reactive oxygen species, and thus provide distinct timescales and mechanisms of protection in a changing environment.

An Arabidopsis thaliana mutant, altered in the γ-subunit of ATP synthase, has a different pattern of intensity-dependent changes in non-photochemical quenching and kinetics of the P-to-S fluorescence decay

2002 ◽  
Vol 29 (4) ◽  
pp. 425 ◽  
Author(s):  
Govindjee ◽  
Paul Spilotro
Keyword(s):  
Arabidopsis Thaliana ◽  
Xanthophyll Cycle ◽  
Fluorescence Decay ◽  
Coupling Factor ◽  
Photochemical Quenching ◽  
Wild Type ◽  
Marked Rise ◽  
Chl A ◽  
Chl A Fluorescence ◽  
Non Photochemical Quenching

A major photoprotective mechanism that plants employ against excess light involves interplay between the xanthophyll cycle and the accumulation of protons. Using mutants in the xanthophyll cycle, the roles of violaxanthin, antheraxanthin and zeaxanthin have already been well established. In this paper, we present data on intact leaves of a mutant [coupling factor quick recovery mutant (cfq); atpC1:E244K] of Arabidopsis thaliana that we expected, based on 515-nm absorbance changes (Gabrys et al. 1994, Plant Physiology 104, 769–776), to have differences in light-induced ΔpH. The significance of this paper is: (i) it is the first study of the photoprotective energy dissipation involving a mutant of the pH gradient; it establishes that protons play an important role in the pattern of non-photochemical quenching (NPQ) of chlorophyll (Chl) a fluorescence; and (ii) differences between the cfq and the wild type (wt) are observed only under subsaturating light intensities, and are strongest in the initial few minutes of the induction period. Our results on light-intensity dependent Chl* a fluorescence transients (the Kautsky effect), and on NPQ of Chl a fluorescence, at 50–250 μmol photons m–2 s–1 demonstrate: (i) the ‘P-to-S’ (or ‘T’) decay, known to be related to [H+] (Briantais et al. 1979, Biochimica et Biophysica Acta 548, 128–138), is slowed in the mutant; and (ii) the pattern of NPQ kinetics is different in the initial 100 s — in the wt leaves, there is a marked rise and decline, and in the cfq mutant, there is a slowed rise. These differences are absent at 750 μmol photons m–2 s–1. Pre-illumination and nigericin (an uncoupler that dissipates the proton gradient) treatment of the cfq mutant, which has lower ΔpH relative to wild type, confirm the conclusion that protons play an important role in the quenching of Chl a fluorescence.

scholarly journals Quantitative Proteomic Analyses Identify STO/BBX24 -Related Proteins Induced by UV-B

2020 ◽  
Vol 21 (7) ◽  
pp. 2496 ◽  
Author(s):  
Guizhen Lyu ◽  
Dongbing Li ◽  
Hui Xiong ◽  
Langtao Xiao ◽  
Jianhua Tong ◽  
...  
Keyword(s):  
Regulatory Network ◽  
Negative Regulator ◽  
Photochemical Quenching ◽  
Absolute Quantitation ◽  
Light Harvesting Complex ◽  
Extra Energy ◽  
In The Wild ◽  
Isobaric Tags ◽  
Non Photochemical Quenching ◽  
Related Proteins

Plants use solar radiation for photosynthesis and are inevitably exposed to UV-B. To adapt to UV-B radiation, plants have evolved a sophisticated strategy, but the mechanism is not well understood. We have previously reported that STO (salt tolerance)/BBX24 is a negative regulator of UV-B-induced photomorphogenesis. However, there is limited knowledge of the regulatory network of STO in UV-B signaling. Here, we report the identification of proteins differentially expressed in the wild type (WT) and sto mutant after UV-B radiation by iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomic analysis to explore differential proteins that depend on STO and UV-B signaling. A total of 8212 proteins were successfully identified, 221 of them were STO-dependent proteins in UV-B irradiated plants. The abundances of STO-dependent PSB and LHC (light-harvesting complex) proteins in sto mutants decreased under UV-B radiation, suggesting that STO is necessary to maintain the normal accumulation of photosynthetic system complex under UV-B radiation to facilitate photosynthesis photon capture. The abundance of phenylalanine lyase-1 (PAL1), chalcone synthetase (CHS), and flavonoid synthetase (FLS) increased significantly after UV-B irradiation, suggesting that the accumulation of flavonoids do not require STO, but UV-B is needed. Under UV-B radiation, STO stabilizes the structure of antenna protein complex by maintaining the accumulation of PSBs and LHCs, thereby enhancing the non-photochemical quenching (NPQ) ability, releasing extra energy, protecting photosynthesis, and ultimately promoting the elongation of hypocotyl. The accumulation of flavonoid synthesis key proteins is independent of STO under UV-B radiation. Overall, our results provide a comprehensive regulatory network of STO in UV-B signaling.

scholarly journals Photosynthesis without β-carotene

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Pengqi Xu ◽  
Volha U Chukhutsina ◽  
Wojciech J Nawrocki ◽  
Gert Schansker ◽  
Ludwik W Bielczynski ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Protein Complexes ◽  
Tobacco Plant ◽  
Oxygenic Photosynthesis ◽  
Photochemical Quenching ◽  
Photoautotrophic Growth ◽  
Pigment Protein Complexes ◽  
Common Opinion ◽  
Non Photochemical Quenching ◽  
Β Carotene

Carotenoids are essential in oxygenic photosynthesis: they stabilize the pigment–protein complexes, are active in harvesting sunlight and in photoprotection. In plants, they are present as carotenes and their oxygenated derivatives, xanthophylls. While mutant plants lacking xanthophylls are capable of photoautotrophic growth, no plants without carotenes in their photosystems have been reported so far, which has led to the common opinion that carotenes are essential for photosynthesis. Here, we report the first plant that grows photoautotrophically in the absence of carotenes: a tobacco plant containing only the xanthophyll astaxanthin. Surprisingly, both photosystems are fully functional despite their carotenoid-binding sites being occupied by astaxanthin instead of β-carotene or remaining empty (i.e. are not occupied by carotenoids). These plants display non-photochemical quenching, despite the absence of both zeaxanthin and lutein and show that tobacco can regulate the ratio between the two photosystems in a very large dynamic range to optimize electron transport.

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