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Tocopherol-deficient rice plants display increased sensitivity to photooxidative stress

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Abstract

Main conclusion

Tocopherol-deficient transgenic plants were generated by silencing HPT and TC. They exhibited distinct phenotype and physiological parameters under high light condition, indicating that tocopherol is indispensable in photoprotection in rice.

Abstract

Tocopherols are lipophilic antioxidants that are synthesized exclusively in photosynthetic organisms. Despite extensive in vivo characterization of tocopherol functions in plants, their functions in the monocot model plant, rice, remain to be determined. In this study, transgenic rice plants constitutively silenced for homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) activity were generated. Silencing of HPT and TC resulted in up to a 98 % reduction in foliar tocopherol content relative to the control plants, which was also confirmed by transcript level analysis. When grown under normal conditions, HPT and TC transgenics showed no distinctive phenotype relative to the control plants, except a slight reduction in plant height and a slight decrease in the first leaf length. However, when exposed to high light at low temperatures, HPT and TC transgenics had a significantly higher leaf yellowing index than the control plants. The tocopherol-deficient plants decreased their total individual chlorophyll levels, their chlorophyll a/b ratio, and the maximum photochemical efficiency of photosystem II, whereas increased lipid peroxidation levels relative to the control plants. Tocopherol deficiency had no effect on ascorbate biosynthesis, but induced glutathione, antheraxanthin, and particularly zeaxanthin biosynthesis for compensation under stressful conditions. However, despite these compensation mechanisms, HPT and TC transgenics still exhibited altered phenotypes under high light at low temperatures. Therefore, it is suggested that tocopherols cannot be replaced and play an indispensable role in photoprotection in rice.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

A:

Antheraxanthin

ABA:

Abscisic acid

ASA:

Ascorbate

chl P:

Geranylgeranyl reductase

Chl:

Chlorophyll

DHA:

Dedydroascorbate

DMBPQ:

2,3-Dimethyl-5-phytyl-1,4-benzoquinone

FW:

Fresh weight

GSH:

Glutathione

GSSG:

Oxidized glutathione

HPT:

Homogentisate phytyltransferase

MDA:

Malondialdehyde

NPQ:

Non-photochemical quenching

NT:

Non transformed plant

PC-8:

Plastochromanol-8

PQ-9:

Plastoquinone-9

PSII:

Photosystem II

ROS:

Reactive Oxygen Species

SXD1:

Sucrose export defective1

TC:

Tocopherol cyclase

TCA:

Trichloroacetic acid

Ubi:

Ubiquitin

V:

Violaxanthin

WT:

Wild type

Z:

Zeaxanthin

References

  • Abbasi AR, Hajirezaei M, Hofius D, Sonnewald U, Voll LM (2007) Specific roles of α- and γ- tocopherol in abiotic stress responses of transgenic tobacco. Plant Physiol 143:1720–1738

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Abbasi AR, Saur A, Hennig P, Tschiersch H, Hajirezaei M, Hofius D, Sonnewald U, Voll LM (2009) Tocopherol deficiency in transgenic tobacco (Nicotiana tabacum L.) plants leads to accelerated senescence. Plant Cell Environ 32:144–157

    Article  CAS  PubMed  Google Scholar 

  • Asada K (1994) Production and action of active oxygen species in photosynthetic tissue. In: Foyer CH, Mullineaux PM (eds) Causes of photooxidative stress and amelioration of defense systems in plants. CRC Press, Boca Raton, pp 77–104

    Google Scholar 

  • Ballottari M, Dall’Osto L, Morosinotto T, Bassi R (2007) Contrasting behavior of higher plant photosystem I and II antenna systems during acclimation. J Biol Chem 282:8947–8958

    Article  CAS  PubMed  Google Scholar 

  • Botha CEJ, Cross RHM, van Bel AJE, Peter CI (2000) Phloem loading in the sucrose-export-defective (SXD-1) mutant maize is limited by callose deposition at plasmodesmata in bundle sheath-vascular parenchyma interface. Protoplasma 214:65–72

    Article  CAS  Google Scholar 

  • Cahoon EB, Hall SE, Ripp KG, Ganzke TS, Hitz WD, Coughlan SJ (2003) Metabolic redesign of vitamin E biosynthesis in plants for tocotrienol production and increased antioxidant content. Nat Biotechnol 21:1082–1087

    Article  CAS  PubMed  Google Scholar 

  • Chi YH, Moon JC, Park JH, Kim HS, Zulfugarov IS, Fanata WI, Jang HH, Lee JR, Lee YM, Kim ST, Chung YY, Lim CO, Kim JY, Yun DJ, Lee CH, Lee KO, Lee SY (2008) Abnormal chloroplast development and growth inhibition in rice thioredoxin m knock-down plants. Plant Physiol 148:808–817

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Collakova E, DellaPenna D (2001) Isolation and functional analysis of homogentisate phytyltransferase from Synechocystis sp. PCC 6803 and Arabidopsis. Plant Physiol 27:1113–1124

    Article  Google Scholar 

  • Collakova E, DellaPenna D (2003) The role of homogentisate phytyltransferase and other tocopherol pathway enzymes in the regulation of tocopherol synthesis during abiotic stress. Plant Physiol 133:930–940

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Dall’Osto L, Holt NE, Kaligotla S, Fuciman M, Cazzaniga S, Carbonera D, Frank HA, Alric J, Bassi R (2012) Zeaxanthin protects plant photosynthesis by modulating chlorophyll triplet yield in specific light-harvesting antenna subunits. J Biol Chem 287:41820–41834

    Article  PubMed Central  PubMed  Google Scholar 

  • Falk J, Munné-Bosch S (2010) Tocochromanol functions in plants: antioxidation and beyond. J Exp Bot 61:1549–1566

    Article  CAS  PubMed  Google Scholar 

  • Foyer CH, Noctor G (2011) Ascorbate and glutathione: the heart of the redox hub. Plant Physiol 155:2–18

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Grasses T, Grimm B, Koroleva O, Jahns P (2001) Loss of α-tocopherol in tobacco plants with decreased geranylgeranyl reductase activity does not modify photosynthesis in optimal growth conditions but increases sensitivity to high-light stress. Planta 213:620–628

    Article  CAS  PubMed  Google Scholar 

  • Havaux M, Lütz C, Grimm B (2003) Chloroplast membrane photostability in chlP transgenic tobacco plants deficient in tocopherols. Plant Physiol 132:300–310

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Havaux M, Eymery F, Porfirova S, Rey P, Dörmann P (2005) Vitamin E protects against photoinhibition and photooxidative stress in Arabidopsis thaliana. Plant Cell 17:3451–3469

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Havaux M, Ksas B, Szewczyk A, Rumeau D, Franck F, Caffarri S, Triantaphylidès C (2009) Vitamin B6 deficient plants display increased sensitivity to high light and photo-oxidative stress. BMC Plant Biol 9:130

    Article  PubMed Central  PubMed  Google Scholar 

  • Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282

    Article  CAS  PubMed  Google Scholar 

  • Höfgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res 16:9877

    Article  PubMed Central  PubMed  Google Scholar 

  • Hofius D, Hajirezaei MR, Geiger M, Tschiersch H, Melzer M, Sonnewald U (2004) RNAi-mediated tocopherol deficiency impairs photoassimilate export in transgenic potato plants. Plant Physiol 135:1256–1268

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Horvath G, Wessjohann L, Bigirimana J, Jansen M, Guisez Y, Caubergs R, Horemans N (2006) Differential distribution of tocopherols and tocotrienols in photosynthetic and non-photosynthetic tissues. Phytochemistry 67:1185–1195

    Article  CAS  PubMed  Google Scholar 

  • Jahns P (1995) The xanthophyll cycle in intermittent light-grown pea plants. Possible functions of chlorophyll a/b-binding proteins. Plant Physiol 108:149–156

    CAS  PubMed Central  PubMed  Google Scholar 

  • Jiang M, Zhang J (2001) Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings. Plant Cell Physiol 42:1265–1273

    Article  CAS  PubMed  Google Scholar 

  • Johnson MP, Davison PA, Ruban AV, Horton P (2008) The xanthophyll cycle pool size controls the kinetics of non-photochemical quenching in Arabidopsis thaliana. FEBS Lett 582:262–266

    Article  CAS  PubMed  Google Scholar 

  • Kanwischer M, Porfirova S, Bergmüller E, Dörmann P (2005) Alterations in tocopherol cyclase activity in transgenic and mutant plants of Arabidopsis affect tocopherol content, tocopherol composition, and oxidative stress. Plant Physiol 137:713–723

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kruk J, Holländer-Czytko H, Oettmeier W, Trebst A (2005) Tocopherol as singlet oxygen scavenger in photosystem II. J Plant Physiol 162:749–757

    Article  CAS  PubMed  Google Scholar 

  • Law MY, Charles SA, Halliwell B (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of paraquat. Biochem J 210:899–903

    CAS  PubMed Central  PubMed  Google Scholar 

  • Li Z, Wakao S, Fischer BB, Niyogi KK (2009) Sensing and responding to excess light. Annu Rev Plant Biol 60:239–260

    Article  CAS  PubMed  Google Scholar 

  • Loh JP, Kiew R, Kee A, Gan LH, Gan Y (1999) Amplified fragment length polymorphism (AFLP) provides molecular markers for the identification of Caladium bicolor cultivars. Ann Bot 84:155–161

    Article  CAS  Google Scholar 

  • Ma J, Takahashi E (1990) Effect of silicon on the growth and phosphorus uptake of rice. Plant Soil 126:115–119

    Article  CAS  Google Scholar 

  • Maeda H, DellaPenna D (2007) Tocopherol functions in photosynthetic organisms. Curr Opin Plant Biol 10:260–265

    Article  CAS  PubMed  Google Scholar 

  • Maeda H, Song W, Sage TL, DellaPenna D (2006) Tocopherols play a crucial role in low-temperature adaptation and phloem loading in Arabidopsis. Plant Cell 18:2710–2732

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Nishiyama Y, Allakhverdiev SI, Murata N (2005) Inhibition of the repair of photosystem II by oxidative stress in cyanobacteria. Photosynth Res 84:1–7

    Article  CAS  PubMed  Google Scholar 

  • Nishiyama Y, Allakhverdiev SI, Murata N (2006) A new paradigm for the action of reactive oxygen species in the photoinhibition of photosystem II. Biochim Biophys Acta 1757:742–749

    Article  CAS  PubMed  Google Scholar 

  • Porfirova S, Bergmuller E, Tropf S, Lemke R, Dörmann P (2002) Isolation of an Arabidopsis mutant lacking vitamin E and identification of a cyclase essential for all tocopherol biosynthesis. Proc Natl Acad Sci USA 99:12495–12500

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394

    Article  CAS  Google Scholar 

  • Sattler SE, Cahoon EB, Coughlan SJ, DellaPenna D (2003) Characterization of tocopherol cyclases from higher plants and cyanobacteria. Evolutionary implications for tocopherol synthesis and function. Plant Physiol 132:2184–2195

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sattler SE, Gilliland LU, Magallanes-Lundback M, Pollard M, DellaPenna D (2004) Vitamin E is essential for seed longevity and for preventing lipid peroxidation during germination. Plant Cell 16:1419–1432

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Semchuk NM, Lushchak OV, Falk J, Krupinska K, Lushchak VI (2009) Inactivation of genes, encoding tocopherol biosynthetic pathway enzymes, results in oxidative stress in outdoor grown Arabidopsis thaliana. Plant Physiol Biochem 47:384–390

    Article  CAS  PubMed  Google Scholar 

  • Shpilyov AV, Zinchenko VV, Shestakov SV, Grimm B, Lokstein H (2005) Inactivation of the geranylgeranyl reductase (ChlP) gene in the cyanobacterium Synechocystis sp. PCC 6803. Biochim Biophys Acta 1706:195–203

    Article  CAS  PubMed  Google Scholar 

  • Streb P, Shang W, Feierabend J, Bligny R (1998) Divergent strategies of photoprotection in high-mountain plants. Planta 207:313–324

    Article  CAS  Google Scholar 

  • Szymańska R, Kruk J (2008) Tocopherol content and isomers’ composition in selected plant species. Plant Physiol Biochem 46:29–33

    Article  PubMed  Google Scholar 

  • Tanaka R, Oster U, Kruse E, Rudiger W, Grimm B (1999) Reduced activity of geranylgeranyl reductase leads to loss of chlorophyll and tocopherol and to partially geranylgeranylated chlorophyll in transgenic tobacco plants expressing antisense RNA for geranylgeranyl reductase. Plant Physiol 120:695–704

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Thayer SS, Björkman O (1990) Leaf xanthophyll content and composition in sun and shade determined by HPLC. Photosynth Res 23:331–343

    Article  CAS  PubMed  Google Scholar 

  • Triantaphylidès C, Havaux M (2009) Singlet oxygen in plants: production, detoxification and signaling. Trends Plant Sci 14:219–228

    Article  PubMed  Google Scholar 

  • Vidi PA, Kanwischer M, Baginsky S, Austin JR, Csucs G, Dörmann P, Kessler F, Bréhélin C (2006) Tocopherol cyclase (VTE1) localization and vitamin E accumulation in chloroplast plastoglobule lipoprotein particles. J Biol Chem 281:11225–11234

    Article  CAS  PubMed  Google Scholar 

  • Yang W, Cahoon RE, Hunter SC, Zhang C, Han J, Borgschulte T, Cahoon EB (2011) Vitamin E biosynthesis: functional characterization of the monocot homogentisate geranylgeranyl transferase. Plant J 65:206–217

    Article  CAS  PubMed  Google Scholar 

  • Zbierzak AM, Kanwischer M, Wille C, Vidi PA, Giavalisco P, Lohmann A, Briesen I, Porfirova S, Bréhélin C, Kessler F, Dörmann P (2009) Intersection of the tocopherol and plastoquinol metabolic pathways at the plastoglobule. Biochem J 425:389–399

    Article  PubMed  Google Scholar 

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Acknowledgments

The research was supported by the grant of the National Natural Science Foundation of China (No. 31070273), the key project of Tianjin Science and Technology Support Program (11ZCGYNC01000) and the 111 Project (No. B08011).

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  1. Haiwei Chen

    Present address: College of Life Sciences, Chifeng College, Chifeng, 024000, China

Authors and Affiliations

  1. Laboratory of Molecular Genetics, College of Life Sciences, Nankai University, Tianjin, 300071, China

    Defu Chen, Haiwei Chen, Luhua Zhang, Xiaoli Shi & Xiwen Chen

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Correspondence to Xiwen Chen.

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Chen, D., Chen, H., Zhang, L. et al. Tocopherol-deficient rice plants display increased sensitivity to photooxidative stress. Planta 239, 1351–1362 (2014). https://doi.org/10.1007/s00425-014-2064-8

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