Genetic Modification of Tomato with the Tobacco Lycopene Β-Cyclase Gene Produces High Β-Carotene and Lycopene Fruit

Total Page:16

File Type:pdf, Size:1020Kb

Genetic Modification of Tomato with the Tobacco Lycopene Β-Cyclase Gene Produces High Β-Carotene and Lycopene Fruit Z. Naturforsch. 2016; 71(9-10)c: 295–301 Louise Ralley, Wolfgang Schucha, Paul D. Fraser and Peter M. Bramley* Genetic modification of tomato with the tobacco lycopene β-cyclase gene produces high β-carotene and lycopene fruit DOI 10.1515/znc-2016-0102 and alleviation of vitamin A deficiency by β-carotene, Received May 18, 2016; revised July 4, 2016; accepted July 6, 2016 which is pro-vitamin A [4]. Deficiency of vitamin A causes xerophthalmia, blindness and premature death, espe- Abstract: Transgenic Solanum lycopersicum plants cially in children aged 1–4 [5]. Since humans cannot expressing an additional copy of the lycopene β-cyclase synthesise carotenoids de novo, these health-promoting gene (LCYB) from Nicotiana tabacum, under the control compounds must be taken in sufficient quantities in the of the Arabidopsis polyubiquitin promoter (UBQ3), have diet. Consequently, increasing their levels in fruit and been generated. Expression of LCYB was increased some vegetables is beneficial to health. Tomato products are 10-fold in ripening fruit compared to vegetative tissues. the most common source of dietary lycopene. Although The ripe fruit showed an orange pigmentation, due to ripe tomato fruit contains β-carotene, the amount is rela- increased levels (up to 5-fold) of β-carotene, with negli- tively low [1]. Therefore, approaches to elevate β-carotene gible changes to other carotenoids, including lycopene. levels, with no reduction in lycopene, are a goal of Phenotypic changes in carotenoids were found in vegeta- plant breeders. One strategy that has been employed to tive tissues, but levels of biosynthetically related isopre- increase levels of health promoting carotenoids in fruits noids such as tocopherols, ubiquinone and plastoquinone and vegetables for human and animal consumption is were barely altered. Transformants showed tolerance to genetic modification [6]. the bleaching herbicide β-cyclase inhibitor, 2-(4-chloro- Lycopene, an acyclic carotenoid and precursor of phenylthio) triethylamine. The phenotype was inherited β-carotene, is formed from the sequential desaturation of for at least three generations. phytoene. These reactions are catalysed by the enzymes Keywords: β-carotene; genetic modification; lycopene; phytoene desaturase (PDS) and ζ-carotene desaturase lycopene β-cyclase; Solanum lycopersicum. (ZDS). The desaturation sequence occurs in the cis geo- metric isomer configuration and the action of a carotene isomerase (CRTISO) converts either cis-neurosporene or Dedication: This work is dedicated to the memory of Professor Peter Böger, who kindly provided the opportunity for PDF and PMB to work poly-cis lycopene to all-trans lycopene prior to cyclisation in his laboratory on the mode of action of bleaching herbicides. [7]. In green tissues, the action of light and chlorophyll are believed to overcome the necessity for CRTISO activ- ity [8]. Cyclisation of lycopene yields β-carotene, via the 1 Introduction action of a β-cyclase. In tomato, two lycopene cyclase are present: LCYB is believed to predominate in the formation Carotenoids are a major group of isoprenoids, widely of vegetative carotenoids, whilst the CYCB gene shows distributed in nature [1]. Several of them are reported to ripening specific expression and CYCB is thus associated have health benefits [2], most notably reduction in the with β-carotene production during ripening [9]. In vegeta- incidence of prostate cancer from dietary lycopene [3] tive tissues, ε-carotene is formed via the action of a ε-ring cyclase and β-ring cyclase, leading to the synthesis of lutein, via hydroxylation, which is the predominant veg- aPresent address: Fraunhofer Chile Research Foundation, Las etative carotenoid [10]. The complete pathway is shown in Condes, Santiago, Chile. Figure 1. *Corresponding author: Peter M. Bramley, School of Biological Since the herbicide 2-(4-chlorophenylthio) triethyl- Sciences, Royal Holloway University of London, Egham, amine (CPTA) is known to inhibit β-carotene formation in Surrey TW20 0EX, UK, E-mail: [email protected] Louise Ralley, Wolfgang Schuch and Paul D. Fraser: School of tomato fruit [11], the transgenic lines have been grown in Biological Sciences, Royal Holloway University of London, Egham, its presence to determine if overexpression of LCYB can Surrey TW20 0EX, UK overcome this inhibition. 296 Ralley et al.: Elevation of β-carotene in tomato fruit by genetic modification CH H3C CH3 3 CH3 x2 GGPP CH3 H3C PSY-1, PSY-2 H3C CH3 H3C CH3 15-cis Phytoene H3C CH3 CH H C 3 3 PDS CH3 H3C H3C CH3 H C CH 9. 15. 9′-tri-cis ζ-Carotene H3C 3 3 CH3 CH3 H3C CH3 H3C ZDS, CRTISO, light CH3 CH3 CH3 CH3 CH3 H3C All-trans Lycopene CH3 CH3 CH3 CH3 LCYE, LCYB LCYB, CYCB H C H C CH 3 H3C 3 CH CH3 3 H C CH CH 3 3 CH3 3 3 α-Carotene β-Carotene H C CH3 CH CH CH CH 3 CH H3C 3 3 3 3 CH3 CH3 3 CRTRB1, CYP97A, CYP97C CRTRB1, CYP97A, ZEP-1, NXS Lutein Zeaxanthin, Antheraxanthin, Violaxanthin, Neoxanthin Figure 1: Carotenoid biosynthesis in tomato. GGPP, geranylgeranyl diphosphate; PSY-1 and -2, phytoene synthase 1 and 2; PDS, phytoene desaturase; ZDS, ζ-carotene desaturase; CRTISO, carotene isomerase; LCYE, lycopene ε-cyclase; LCYB, lycopene β-cyclase; CYCB, fruit specific lycopene β-cyclase; CRTR-B1, β-carotene hydroxylase; CYP97A, P450 carotenoid β-ring hydroxylase; CYP97C, P450 carotenoid ε-hydroxylase; ZEP-1, zeaxanthin epoxi- dase; NXS, neoxanthin synthase. 2 Materials and methods the pSP vector (Promega, Southampton, UK), containing the polyubiquitin promoter (UBQ3; At5g03240; UniProt KB-Q1EC66) from Arabidopsis thaliana, the nopaline syn- 2.1 Plant material thase terminator, the nptII gene for kanamycin resistance, controlled by the AoPR1 promoter from Asparagus offici- Tomato (Solanum lycopersicum cv Ailsa Craig) plants were nalis. The Age1 fragment was sub-cloned into the pVB6 grown in the glasshouse with supplementary lighting. plant binary vector and designated pVBLCYB (Figure 2). Three plants per genotype were grown in a randomised The vector/insert was sequenced to confirm the manipula- manner concurrently with their respective backgrounds tions. E. coli strain XL1-Blue (Promega) was used in these and fruit harvested at mature red ripe (7 days post breaker, experiments. Sequences of the tobacco and tomato LCYB dpb). Two or more fruits per plant were pooled to provide are shown in Supplementary Material, Figure 1. one replicate per plant and three per genotype. 2.3 Transformation of tomato plants 2.2 Construction of the pVBLCYB vector Agrobacterium tumifaciens LBA 4404 was transformed Digestion of pUC19 with Sma1 released the LCYB cDNA as with pVBLCYB by triparental mating, using the helper a 1.6-kb fragment, including the sequence encoding the plasmid pRK2103 [12]. Stem explants of tomato were trans- transit peptide, which was cloned into the Sma1 site of formed and plants regenerated as described previously Ralley et al.: Elevation of β-carotene in tomato fruit by genetic modification 297 pVBLCY Age 1 Asc1 BamH1 Asc1 Age 1 UBQ3 LB AoPR1 nptIInos UBQ3 LYCB nonossRB Figure 2: Schematic diagram of the LCYB construct pVBLCY. UBQ3, ubiquitin three promotor; LCYB, N. tabacum lycopene β-cyclase; nptII, nopaline synthase terminator; ASe1, BamH1, Age1 restriction sites; LB, left border; RB, right border. [13]. Kanamycin (50 mg/ml) and carbenicillin (250 mg/ml) 2-(4-chlorophenylthio) triethylamine (CPTA, a gift from were used to select resistant transformants. Prof. P. Böger) at 30 and 40 μM. The seedlings were grown in a controlled environment with a 16-h photoperiod and day and night temperatures of 23 and 19°C, respectively. 2.4 DNA and RNA analyses They were assessed for germination and bleaching after 2 weeks and samples taken for isoprenoid analysis. DNA was extracted for PCR analysis from leaf material (ca. 200 mg), as described previously [13]. The presence of the transgene in the primary transformants (T0) was 2.6 Analysis of isoprenoids confirmed by PCR using primers designed to amplify a 500 bp internal fragment of LCYB. Southern blot analy- The isoprenoids of T0, T1 and T2 progeny of leaf and 7 dpb sis of the T0 and first-generation (T1) plants was carried fruit were extracted, separated, identified and quantified out using the cetyltrimethylammonium bromide (CTAB) by HPLC analyses, as described in [16]. method [14]. DNA was digested with the appropriate restriction enzymes, fragments separated in 1% (w/v) agarose gels and transferred to Hybond N+ (Amersham, Little Chalfont, UK) overnight in 20% saline sodium 3 Results citrate (SSC) solution. 32P-labelled fragments (using the Stratagene Random primer labelling kit, Prime It II), cor- 3.1 Plants expressing the LCYB gene from responding to the LCYB and nptII regions of the transgene, tobacco showed increased levels of were used to probe the filters. Hybridisation, washing and β + -carotene in ripe fruit and changes to detection were performed as described in the Hybond N leaf carotenoids instructions. Northern blot analysis was carried out on tomato fruit of the T and T generations, harvested at 7 dpb o 1 Following transformation, progeny were selected on the [15]. Total RNA (20 μg) from the fruit was electrophoresed basis of kanamycin resistance (nptII) and PCR analysis. on 1.4% agarose/formaldehyde gels and then transferred During regeneration, growth and development of veg- to Hybond-N+ in 20x SSC overnight. The 1.6-kb cDNA frag- etative tissues from all transgenic lines appeared pheno- ment was radiolabelled with 32P, as described above. Fol- typically normal. However, as fruit ripening commenced, lowing pre-hybridisation, the filter was hybridised at 65°C virtually all transgenic lines showed a greater degree of in 7.5% SDS, 100 mM K phosphate buffer, pH 7.5. Each filter orange colouration in their fruit compared to the control. was washed for 15 min in 2× SSC, 0.5% SDS and then twice Analysis of the T generation showed that increases in for 15 min with 1× SSC, 0.5% SDS and finally monitored 0 β-carotene content of the fruit were responsible for the for radioactivity for 24 h with a phosphorimager (MAcBAS altered colouration (Table 1).
Recommended publications
  • Journal of Nephropathology
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by shahrekord university of medical scinces www.nephropathol.com DOI: 10.15171/jnp.2017.25 J Nephropathol. 2017;6(3):144-149 Journal of Nephropathology Ameliorative effect of lycopene effect on cisplatin-induced nephropathy in patients Leila Mahmoodnia, Keivan Mohammadi*, Rohollah Masumi Department of Internal Medicine, Shahrekord University of Medical Sciences, Shahrekord, Iran ARTICLE INFO ABSTRACT Article type: Background: Nephrotoxicity is one of the most important limitations of cisplatin-based Original Article chemotherapies which associated with many complications and high mortality rate. Objectives: To investigate the effect of lycopene on cisplatin-induced nephrotoxicity in Article history: patients with cancer. Received: 14 November 2016 Accepted: 2 January 2017 Patients and Methods: In this double-blind, randomized clinical trial, 120 patients were Published online: 17 January 2017 randomly assigned to two groups, case (treated with lycopene + standard regimen of kidney DOI: 10.15171/jnp.2017.25 injury prevention) and control (treated with only the standard regimen of kidney injury prevention). Lycopene was orally taken from 24 hours before to 72 hours after cisplatin Keywords: administration. Blood urea nitrogen (BUN), serum creatinine (Cr), and glomerular Lycopene filtration rate (GFR) were measured and recorded. The data were analyzed using SPSS. Nephrotoxicity Results: Changes in Cr were not significantly different between the two groups (P = 0.131). Cisplatin However, a significant decreasing trend was seen in GFR during the study, which was Cancer more marked in the control group (P = 0.004). BUN significantly decreased during the Original Article study (P = 0.002), and a significant decrease of BUN on the day three in both groups was seen (P = 0.001).
    [Show full text]
  • Altered Xanthophyll Compositions Adversely Affect Chlorophyll Accumulation and Nonphotochemical Quenching in Arabidopsis Mutants
    Proc. Natl. Acad. Sci. USA Vol. 95, pp. 13324–13329, October 1998 Plant Biology Altered xanthophyll compositions adversely affect chlorophyll accumulation and nonphotochemical quenching in Arabidopsis mutants BARRY J. POGSON*, KRISHNA K. NIYOGI†,OLLE BJO¨RKMAN‡, AND DEAN DELLAPENNA§¶ *Department of Plant Biology, Arizona State University, Tempe, AZ 85287-1601; †Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102; ‡Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA 94305-4101; and §Department of Biochemistry, University of Nevada, Reno, NV 89557-0014 Contributed by Olle Bjo¨rkman, September 4, 1998 ABSTRACT Collectively, the xanthophyll class of carote- thin, are enriched in the LHCs, where they contribute to noids perform a variety of critical roles in light harvesting assembly, light harvesting, and photoprotection (2–8). antenna assembly and function. The xanthophyll composition A summary of the carotenoid biosynthetic pathway of higher of higher plant photosystems (lutein, violaxanthin, and neox- plants and relevant chemical structures is shown in Fig. 1. anthin) is remarkably conserved, suggesting important func- Lycopene is cyclized twice by the enzyme lycopene b-cyclase tional roles for each. We have taken a molecular genetic to form b-carotene. The two beta rings of b-carotene are approach in Arabidopsis toward defining the respective roles of subjected to identical hydroxylation reactions to yield zeaxan- individual xanthophylls in vivo by using a series of mutant thin, which in turn is epoxidated once to form antheraxanthin lines that selectively eliminate and substitute a range of and twice to form violaxanthin. Neoxanthin is derived from xanthophylls. The mutations, lut1 and lut2 (lut 5 lutein violaxanthin by an additional rearrangement (9).
    [Show full text]
  • The Biosynthesis of Carotenoids
    THE BIOSYNTHESIS OF CAROTENOIDS C. 0. CHICHESTER Department of Food Science and Technology, University of a4fornia, Davis, Galjf 95616, U.S.A. INTRODUCTION Considerable progress has been made in the field of carotenoid bio- chemistry in the last ten to fifteen years. Prior to that very little, if anything, was known about the units which formed the intermediates of the coloured 40—carbon pigments, or the structures of those that were proposed as intermediates. Like all fields of biochemistry, as knowledge is accumulated specific problems are crystallized. These generally concern key areas, which until an understanding is achieved can substantially hinder the unification of a field or problem. The biochemistry of the carotenoids has arrived at this point. INTERMEDIATE COMPOUNDS IN CAROTENOID BIOSYNTHESIS There is general consensus as to the identity of the intermediate com- pounds which form the building blocks of the carotenoids. Studies in Phycornyces blakesleeanus, tomatoes, corn, carrots, Mucor hiemalis, spinach leaves, and bean leaves all coincide to indicate that the common building block of carotenoids is mevalonic acid'—7. Some experiments concerned with carotenoids, and many with sterols, have shown that /3-methyl-/3- hydroxyglutarate-CoA, and/or acetoacetic-CoA, are the normal sources of inevalonic acid" 8 In Phycomyces blalcesleeanus, as well as tomatoes, it has been shown rather unequivocally that the first steps in the conversion of meva- lonic acid to the C—40's are identical to those postulated for the formation of sterols911. The mevalonic acid is converted via the 5-phosphomevalonic acid to the 5-pyrophosphoryl mevalonic acid'2. This compound is then decarboxylated to form the z3-isopentenol pyrophosphate.
    [Show full text]
  • Identification of Distinct Ph- and Zeaxanthin-Dependent Quenching
    RESEARCH ARTICLE Identification of distinct pH- and zeaxanthin-dependent quenching in LHCSR3 from Chlamydomonas reinhardtii Julianne M Troiano1†, Federico Perozeni2†, Raymundo Moya1, Luca Zuliani2, Kwangyrul Baek3, EonSeon Jin3, Stefano Cazzaniga2, Matteo Ballottari2*, Gabriela S Schlau-Cohen1* 1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States; 2Department of Biotechnology, University of Verona, Verona, Italy; 3Department of Life Science, Hanyang University, Seoul, Republic of Korea Abstract Under high light, oxygenic photosynthetic organisms avoid photodamage by thermally dissipating absorbed energy, which is called nonphotochemical quenching. In green algae, a chlorophyll and carotenoid-binding protein, light-harvesting complex stress-related (LHCSR3), detects excess energy via a pH drop and serves as a quenching site. Using a combined in vivo and in vitro approach, we investigated quenching within LHCSR3 from Chlamydomonas reinhardtii. In vitro two distinct quenching processes, individually controlled by pH and zeaxanthin, were identified within LHCSR3. The pH-dependent quenching was removed within a mutant LHCSR3 that lacks the residues that are protonated to sense the pH drop. Observation of quenching in zeaxanthin-enriched LHCSR3 even at neutral pH demonstrated zeaxanthin-dependent quenching, which also occurs in other light-harvesting complexes. Either pH- or zeaxanthin-dependent quenching prevented the formation of damaging reactive oxygen species, and thus the two *For correspondence: quenching processes may together provide different induction and recovery kinetics for [email protected] (MB); photoprotection in a changing environment. [email protected] (GSS-C) †These authors contributed equally to this work Competing interests: The Introduction authors declare that no Sunlight is the essential source of energy for most photosynthetic organisms, yet sunlight in excess competing interests exist.
    [Show full text]
  • Synthetic Conversion of Leaf Chloroplasts Into Carotenoid-Rich Plastids Reveals Mechanistic Basis of Natural Chromoplast Development
    Synthetic conversion of leaf chloroplasts into carotenoid-rich plastids reveals mechanistic basis of natural chromoplast development Briardo Llorentea,b,c,1, Salvador Torres-Montillaa, Luca Morellia, Igor Florez-Sarasaa, José Tomás Matusa,d, Miguel Ezquerroa, Lucio D’Andreaa,e, Fakhreddine Houhouf, Eszter Majerf, Belén Picóg, Jaime Cebollag, Adrian Troncosoh, Alisdair R. Ferniee, José-Antonio Daròsf, and Manuel Rodriguez-Concepciona,f,1 aCentre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB, Campus UAB Bellaterra, 08193 Barcelona, Spain; bARC Center of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney NSW 2109, Australia; cCSIRO Synthetic Biology Future Science Platform, Sydney NSW 2109, Australia; dInstitute for Integrative Systems Biology (I2SysBio), Universitat de Valencia-CSIC, 46908 Paterna, Valencia, Spain; eMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany; fInstituto de Biología Molecular y Celular de Plantas, CSIC-Universitat Politècnica de València, 46022 Valencia, Spain; gInstituto de Conservación y Mejora de la Agrodiversidad, Universitat Politècnica de València, 46022 Valencia, Spain; and hSorbonne Universités, Université de Technologie de Compiègne, Génie Enzymatique et Cellulaire, UMR-CNRS 7025, CS 60319, 60203 Compiègne Cedex, France Edited by Krishna K. Niyogi, University of California, Berkeley, CA, and approved July 29, 2020 (received for review March 9, 2020) Plastids, the defining organelles of plant cells, undergo physiological chromoplasts but into a completely different type of plastids and morphological changes to fulfill distinct biological functions. In named gerontoplasts (1, 2). particular, the differentiation of chloroplasts into chromoplasts The most prominent changes during chloroplast-to-chromo- results in an enhanced storage capacity for carotenoids with indus- plast differentiation are the reorganization of the internal plastid trial and nutritional value such as beta-carotene (provitamin A).
    [Show full text]
  • Meet Lycopene Prostate Cancer Is One of the Leading Causes of Cancer Death Among Men in the United States
    UCLA Nutrition Noteworthy Title Lycopene and Mr. Prostate: Best Friends Forever Permalink https://escholarship.org/uc/item/5ks510rw Journal Nutrition Noteworthy, 5(1) Author Simzar, Soheil Publication Date 2002 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Meet Lycopene Prostate cancer is one of the leading causes of cancer death among men in the United States. Dietary factors are considered an important risk factor for the development of prostate cancer in addition to age, genetic predisposition, environmental factors, and other lifestyle factors such as smoking. Recent studies have indicated that there is a direct correlation between the occurrence of prostate cancer and the consumption of tomatoes and tomato-based products. Lycopene, one of over 600 carotenoids, is one of the main carotenoids found in human plasma and it is responsible for the red pigment found in tomatoes and other foods such as watermelons and red grapefruits. It has been shown to be a very potent antioxidant, with oxygen-quenching ability greater than any other carotenoid. Recent research has indicated that its antioxidant effects help lower the risk of heart disease, atherosclerosis, and different types of cancer-especially prostate cancer. Lycopene's Characteristics Lycopene is on of approximately 600 known carotenoids. Carotenoids are red, yellow, and orange pigments which are widely distributed in nature and are especially abundant in yellow- orange fruits and vegetables and dark green, leafy vegetables. They absorb light in the 400- 500nm region which gives them a red/yellow color. Only green plants and certain microorganisms such as fungi and algae can synthesize these pigments.
    [Show full text]
  • Lycopene Analysis and Horticultural Attributes of Tomatoes. MS Thesis
    THESIS LYCOPENE ANALYSIS AND HORTICULTURAL ATTRIBUTES OF TOMATOES Submitted by Samuel E. Cox Department of Horticulture In partial fulfillment of the requirements for the Degree of Master of Science Colorado State University Fort Collins, Colorado Spring 2001 COLORADO STATE UNIVERSITY March 9, 2001 WE HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER OUR SUPERVISION BY SAMUEL E. COX ENTITLED LYCOPENE ANALYSIS AND HORTICULTURAL ATTRIBUTES OF TOMATOES BE ACCEPTED AS FULFILLING IN PART REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE Committee on Graduate Work Michelle L. Jones David A. Sampson Cecil Stushnoff Advisor Steve Wallner Department Head ii THESIS ABSTRACT LYCOPENE ANALYSIS AND HORTICULTURAL ATTRIBUTES OF TOMATOES Tomatoes are the most popular home garden vegetables grown in the United States, and have one of the highest value/acre ratios of any commercially produced crop. Popularity of tomatoes, combined with competition among seed companies, have led to an abundance and often bewildering variety of cultivars intended for home garden cultivation. Thirty home garden cultivars marketed at local nurseries in northern Colorado were grown and evaluated. Plant health, morphology and size, subjective plant yield observations and objective taste measurements were all investigated in order to identify exceptional or unacceptable cultivars for this area. Cellular damage from free radicals is a major cause of degenerative diseases and cancer. Lycopene is an open-chain hydrocarbon carotenoid found in abundance in tomatoes that has been shown to possess strong antioxidant activity in animal systems. This led to the discovery that several types of cancers are inhibited by its consumption. Research into the properties and health benefits of lycopene has been a relatively recent development and will undoubtedly continue to grow.
    [Show full text]
  • Identification of 1 Distinct Ph- and Zeaxanthin-Dependent Quenching in LHCSR3 from Chlamydomonas Reinhardtii
    Identification of 1 distinct pH- and zeaxanthin-dependent quenching in LHCSR3 from chlamydomonas reinhardtii The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Troiano, Julianne M. et al. “Identification of 1 distinct pH- and zeaxanthin-dependent quenching in LHCSR3 from chlamydomonas reinhardtii.” eLife, 10 (January 2021): e60383 © 2021 The Author(s) As Published 10.7554/eLife.60383 Publisher eLife Sciences Publications, Ltd Version Final published version Citable link https://hdl.handle.net/1721.1/130449 Terms of Use Creative Commons Attribution 4.0 International license Detailed Terms https://creativecommons.org/licenses/by/4.0/ RESEARCH ARTICLE Identification of distinct pH- and zeaxanthin-dependent quenching in LHCSR3 from Chlamydomonas reinhardtii Julianne M Troiano1†, Federico Perozeni2†, Raymundo Moya1, Luca Zuliani2, Kwangyrul Baek3, EonSeon Jin3, Stefano Cazzaniga2, Matteo Ballottari2*, Gabriela S Schlau-Cohen1* 1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States; 2Department of Biotechnology, University of Verona, Verona, Italy; 3Department of Life Science, Hanyang University, Seoul, Republic of Korea Abstract Under high light, oxygenic photosynthetic organisms avoid photodamage by thermally dissipating absorbed energy, which is called nonphotochemical quenching. In green algae, a chlorophyll and carotenoid-binding protein, light-harvesting complex stress-related (LHCSR3), detects excess energy via a pH drop and serves as a quenching site. Using a combined in vivo and in vitro approach, we investigated quenching within LHCSR3 from Chlamydomonas reinhardtii. In vitro two distinct quenching processes, individually controlled by pH and zeaxanthin, were identified within LHCSR3. The pH-dependent quenching was removed within a mutant LHCSR3 that lacks the residues that are protonated to sense the pH drop.
    [Show full text]
  • Abscisic Acid Induced Protection Against Photoinhibition of PSII Correlates with Enhanced Activity of the Xanthophyll Cycle
    FEBS 15944 FEBS Letters 371 (1995) 61-64 Abscisic acid induced protection against photoinhibition of PSII correlates with enhanced activity of the xanthophyll cycle A.G. Ivanov a'*, M. Krol b, D. Maxwell b, N.P.A. Huner b alnstitute of Biophysics, Bulgarian Academy of Sciences, Aead. (7. Bonchev Street, bl. 21, 1113 Sofia, Bulgaria bDepartment of Plant Sciences, University of Western Ontario, London, Ont. N6A 5B7, Canada Received 22 June 1995; revised version received 24 July 1995 applied ABA on the light-dependent zeaxanthin formation, the Abstract The exogenous application of abscisic acid (ABA) to related capacity for non-photochemical chlorophyll fluores- barley seedlings resulted in partial protection of the PSII photo- cence quenching and the possible involvement of ABA in the chemistry against photoinhibition at low temperature, the effect protection of PSII photochemistry from excessive radiation at being most pronounced at 10 -s M ABA. This was accompanied low temperatures. by higher photochemical quenching (qP) in ABA-treated leaves. A considerable increase (122%) in the amount of total carotenoids 2. Materials and methods and xanthophylls (antheraxanthin, violaxanthin and zeaxanthin) was also found in the seedlings subjected to ABA. The activity Seeds of barley (Hordeum vulgare L. var. cadette) were germinated of the xanthophyll cycle measured by the epoxidation state of for 3 days and grown in aqueous solutions of ABA (10 -5 M and 10-6 M) xanthophyUs under high-light treatment was higher in ABA- as in [23]. ABA treatments lasted 7 days. The solutions were changed treated plants compared with the control. This corresponds to a daily.
    [Show full text]
  • Carotene and Lycopene Isoforms, and of Antioxidant Potential in Human Blood Bioavailability: a Pilot Study
    nutrients Article Nutritional Controlled Preparation and Administration of Different Tomato Purées Indicate Increase of β-Carotene and Lycopene Isoforms, and of Antioxidant Potential in Human Blood Bioavailability: A Pilot Study Daniela Vitucci 1,†, Angela Amoresano 2,†, Marcella Nunziato 1,3,† , Simona Muoio 4, Andreina Alfieri 1,5 , Giovannangelo Oriani 1, Luca Scalfi 6, Luigi Frusciante 7, Maria Manuela Rigano 7 , Piero Pucci 1,2, Luigi Fontana 8,9,10, Pasqualina Buono 1,5,* and Francesco Salvatore 1,3,* 1 CEINGE-Biotecnologie Avanzate, Via G. Salvatore, 486, 80145 Naples, Italy; [email protected] (D.V.); [email protected] (M.N.); andreina.alfi[email protected] (A.A.); [email protected] (G.O.); [email protected] (P.P.) 2 Department of Chemical Sciences, University of Naples “Federico II”, Via Cinthia, 80126 Naples, Italy; [email protected] 3 Department of Molecular Medicine and Medical Biotechnologies, University of Naples “Federico II”, Via Sergio Pansini 5, 80131 Naples, Italy 4 Department of Public Health, School of Medicine, University of Naples “Federico II”, 80131 Naples, Italy; [email protected] 5 Department of Human Movement Sciences and Wellbeing, University of Naples “Parthenope”, Via Medina, Citation: Vitucci, D.; Amoresano, A.; 40, 80133 Naples, Italy Nunziato, M.; Muoio, S.; Alfieri, A.; 6 Institute of Internal Medicine and Metabolic Diseases, Medical School, University of Naples, Federico II, Oriani, G.; Scalfi, L.; Frusciante, L.; 80131 Naples, Italy; scalfi@unina.it Rigano, M.M.; Pucci, P.; et al. 7 Department
    [Show full text]
  • Carotenoid Composition of Strawberry Tree (Arbutus Unedo L.) Fruits
    Accepted Manuscript Carotenoid composition of strawberry tree (Arbutus unedo L.) fruits Raúl Delgado-Pelayo, Lourdes Gallardo-Guerrero, Dámaso Hornero-Méndez PII: S0308-8146(15)30273-9 DOI: http://dx.doi.org/10.1016/j.foodchem.2015.11.135 Reference: FOCH 18476 To appear in: Food Chemistry Received Date: 25 May 2015 Revised Date: 21 November 2015 Accepted Date: 28 November 2015 Please cite this article as: Delgado-Pelayo, R., Gallardo-Guerrero, L., Hornero-Méndez, D., Carotenoid composition of strawberry tree (Arbutus unedo L.) fruits, Food Chemistry (2015), doi: http://dx.doi.org/10.1016/j.foodchem. 2015.11.135 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Carotenoid composition of strawberry tree (Arbutus unedo L.) fruits. Raúl Delgado-Pelayo, Lourdes Gallardo-Guerrero, Dámaso Hornero-Méndez* Group of Chemistry and Biochemistry of Pigments. Food Phytochemistry Department. Instituto de la Grasa (CSIC). Campus Universidad Pablo de Olavide, Ctra. de Utrera km. 1. 41013 - Sevilla (Spain). * Corresponding author. Telephone: +34 954611550; Fax: +34 954616790; e-mail: [email protected] 1 Abstract The carotenoid composition of strawberry tree (A. unedo) fruits has been characterised in detail and quantified for the first time. According to the total carotenoid content (over 340 µg/g dw), mature strawberry tree berries can be classified as fruits with very high carotenoid content (> 20 µg/g dw).
    [Show full text]
  • Enhanced Lycopene Production in Escherichia Coli by Expression of Two MEP Pathway Enzymes from Vibrio Sp
    catalysts Article Enhanced Lycopene Production in Escherichia coli by Expression of Two MEP Pathway Enzymes from Vibrio sp. Dhg 1, 1, 1 1, Min Jae Kim y, Myung Hyun Noh y , Sunghwa Woo , Hyun Gyu Lim * and Gyoo Yeol Jung 1,2,* 1 Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Korea; [email protected] (M.J.K.); [email protected] (M.H.N.); [email protected] (S.W.) 2 School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Korea * Correspondence: [email protected] (H.G.L.); [email protected] (G.Y.J.); Tel.: +82-54-279-2391 (G.Y.J.) These authors contributed equally to this work. y Received: 28 October 2019; Accepted: 26 November 2019; Published: 29 November 2019 Abstract: Microbial production is a promising method that can overcome major limitations in conventional methods of lycopene production, such as low yields and variations in product quality. Significant efforts have been made to improve lycopene production by engineering either the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway or mevalonate (MVA) pathway in microorganisms. To further improve lycopene production, it is critical to utilize metabolic enzymes with high specific activities. Two enzymes, 1-deoxy-d-xylulose-5-phosphate synthase (Dxs) and farnesyl diphosphate synthase (IspA), are required in lycopene production using MEP pathway. Here, we evaluated the activities of Dxs and IspA of Vibrio sp. dhg, a newly isolated and fast-growing microorganism.
    [Show full text]