Plant Molecular Biology

, Volume 75, Issue 3, pp 211–221

ZEBRA2, encoding a carotenoid isomerase, is involved in photoprotection in rice

  • Chenglin Chai
  • Jun Fang
  • Yang Liu
  • Hongning Tong
  • Yanqing Gong
  • Yiqin Wang
  • Min Liu
  • Youping Wang
  • Qian Qian
  • Zhukuan Cheng
  • Chengcai Chu


zebra” mutants have alternating green and chlorotic crossbands on leaf blades and are widely distributed in monocotyledonous crops. Most recently, we cloned the first responsible gene from rice, ZEBRA2, which also leads to the phenotype of rice preharvest sprouting. ZEBRA2, a single-copy gene in the rice genome, encodes a carotenoid isomerase (CRTISO), the key enzyme catalyzing the conversion of cis-lycopene to all-trans lycopene. ZEBRA2 shares high identity with known CRTISOs from other species. Expression analysis via both RT-PCR and ZEBRA2-promoter-β-glucuronidase (GUS) transgenic rice indicates that ZEBRA2 is predominantly expressed in mesophyll cells of mature leaves where active photosynthesis occurs. Consistent with the alteration in agronomic traits, the zebra2 mutant exhibits decreased photosynthetic rate and chlorophyll content. Mutation of the ZEBRA2 gene results in the accumulation of all-trans-lycopene precursor, prolycopene (7Z,9Z,7′Z,9′Z tetra cis-lycopene), in dark-grown zebra2 tissues. Light-grown zebra2 mutant exhibits the characteristic “zebra” phenotype and decreased level of lutein, the xanthophyll that is essential for efficient chl triplet quenching. More severe phenotype of the zebra2 mutant under high light intensity indicates that “zebra” phenotype might be caused by photooxidative damages. We conclude that ZEBRA2 is involved in photoprotection in rice.


Carotenoid biosynthesis Carotenoid isomerase Rice zebra2 mutant 

Supplementary material

11103_2010_9719_MOESM1_ESM.doc (64 kb)
Supplementary material 1 (DOC 64 kb)
11103_2010_9719_MOESM2_ESM.jpg (451 kb)
Supplementary material 2 (JPEG 451 kb)
11103_2010_9719_MOESM3_ESM.tif (4.9 mb)
Supplementary material 3 (TIFF 4972 kb)
11103_2010_9719_MOESM4_ESM.tif (2 mb)
Supplementary material 4 (TIFF 2026 kb)
11103_2010_9719_MOESM5_ESM.tif (1 mb)
Supplementary material 5 (TIFF 1035 kb)


  1. Aluru MR, Rodermel SR (2004) Control of chloroplast redox by the IMMUTANS terminal oxidase. Physiol Plant 120:4–11CrossRefPubMedGoogle Scholar
  2. Arnon D (1949) Copper enzymes in isolated chloroplasts: polyphenoloxidase in Beta vulgaris. Plant Physiol 24:15CrossRefGoogle Scholar
  3. Bartley GE, Scolnik PA (1995) Plant carotenoids: pigments for photoprotection, visual attraction, and human health. Plant Cell 7:1027–1038CrossRefPubMedGoogle Scholar
  4. Breitenbach J, Vioque A, Sandmann G (2001) Gene sll0033 from Synechocystis 6803 encodes a carotene isomerase involved in the biosynthesis of all-E lycopene. Z Naturforsch 56:915–917Google Scholar
  5. Carol P, Stevenson D, Bisanz C et al (1999) Mutations in the Arabidopsis gene IMMUTANS cause a variegated phenotype by inactivating a chloroplast terminal oxidase associated with phytoene desaturation. Plant Cell 11:57–68CrossRefPubMedGoogle Scholar
  6. Chen M, Choi Y, Voytas DF, Rodermel S (2000) Mutations in the Arabidopsis VAR2 locus cause leaf variegation due to the loss of a chloroplast FtsH protease. Plant J 22:303–313CrossRefPubMedGoogle Scholar
  7. Chen Y, Li F, Wurtzel ET (2010) Isolation and characterization of the Z-ISO gene encoding a missing component of carotenoid biosynthesis in plants. Plant Physiol 153:66–79CrossRefPubMedGoogle Scholar
  8. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159CrossRefPubMedGoogle Scholar
  9. Coe EH, Hoisington DA, Neuffer MG (1987) Linkage map of corn (maize). MNL 61:116–123Google Scholar
  10. Conti A, Pancaldi S, Fambrini M, Michelotti V, Bonora A, Salvini M, Pugliesi C (2004) A deficiency at the gene coding for zeta-carotene desaturase characterizes the sunflower non dormant-1 mutant. Plant Cell Physiol 45:445–455CrossRefPubMedGoogle Scholar
  11. Cunningham FX, Gantt E (1998) Genes and enzymes of carotenoid biosynthesis in plants. Annu Rev Plant Physiol Plant Mol Biol 49:557–583CrossRefPubMedGoogle Scholar
  12. Dall’Osto L, Lico C, Alric J, Giuliano G, Havaux M, Bassi R (2006) Lutein is needed for efficient chlorophyll triplet quenching in the major LHCII antenna complex of higher plants and effective photoprotection in vivo under strong light. BMC Plant Biol 6:32CrossRefPubMedGoogle Scholar
  13. Demmig-Adams B, Adams WW III (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1:21–26CrossRefGoogle Scholar
  14. Dong H, Deng Y, Mu J et al (2007) The Arabidopsis Spontaneous Cell Death1 gene, encoding a zeta-carotene desaturase essential for carotenoid biosynthesis, is involved in chloroplast development, photoprotection and retrograde signalling. Cell Res 17:458–470PubMedGoogle Scholar
  15. Fang J, Chai C, Qian Q et al (2008) Mutations of genes in synthesis of the carotenoid precursors of ABA lead to pre-harvest sprouting and photo-oxidation in rice. Plant J 54:177–189CrossRefPubMedGoogle Scholar
  16. Fong F, Smith JD, Koehler DE (1983) Early events in maize seed development: 1-methyl-3-phenyl-5-(3-[trifluoromethyl]phenyl)-4-(1H)-pyridinone induction of vivipary. Plant Physiol 73:899–901CrossRefPubMedGoogle Scholar
  17. Formaggio E, Cinque G, Bassi R (2001) Functional architecture of the major light-harvesting complex from higher plants. J Mol Biol 314:1157–1166CrossRefPubMedGoogle Scholar
  18. Frank HA, Cogdell RJ (1996) Carotenoids in photosynthesis. Photochem Photobiol 63:257–264CrossRefPubMedGoogle Scholar
  19. Fraser PD, Pinto ME, Holloway DE, Bramley PM (2000) Technical advance: application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids. Plant J 24:551–558CrossRefPubMedGoogle Scholar
  20. Giraudat J, Parcy F, Bertauche N et al (1994) Current advances in abscisic acid action and signalling. Plant Mol Biol 26:1557–1577CrossRefPubMedGoogle Scholar
  21. Groot S, Karssen C (1992) Dormancy and germination of abscisic acid-deficient tomato seeds: studies with the sitiens mutant. Plant Physiol 99:952–958CrossRefPubMedGoogle Scholar
  22. Hable WE, Oishi KK, Schumaker KS (1998) Viviparous-5 encodes phytoene desaturase, an enzyme essential for abscisic acid (ABA) accumulation and seed development in maize. Mol Gen Genet 257:167–176CrossRefPubMedGoogle Scholar
  23. Havaux M (1998) Carotenoids as membrane stabilizers in chloroplasts. Trends Plant Sci 3:147–151CrossRefGoogle Scholar
  24. Hayes HK (1932) Zebra crossbands. J Hered 23:415–419Google Scholar
  25. He R, Ding Y, Yu J, Gao W (2000) The changes of chlorophyll content and several enzyme activities in zebra-leaf rice. J Wuhan Univer (Nat Sci Ed) 46:761–765Google Scholar
  26. Isaacson T, Ronen G, Zamir D, Hirschberg J (2002) Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of beta-carotene and xanthophylls in plants. Plant Cell 14:333–342CrossRefPubMedGoogle Scholar
  27. Isaacson T, Ohad I, Beyer P, Hirschberg J (2004) Analysis in vitro of the enzyme CRTISO establishes a poly-cis-carotenoid biosynthesis pathway in plants. Plant Physiol 136:4246–4255CrossRefPubMedGoogle Scholar
  28. Janick-Buckner D, O’Neal J, Joyce E, Bucker B (2001) Genetic and biochemical analysis of the y9 gene of maize, a carotenoid biosynthetic gene. Maydica 46:41–46Google Scholar
  29. Jefferson RA (1989) The GUS reporter gene system. Nature 342:837–838CrossRefPubMedGoogle Scholar
  30. Kadam BS, Patel SM, Kalkarni RK (1940) Consequences of in-breeding in bajra. J Hered 31:201–207Google Scholar
  31. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163CrossRefPubMedGoogle Scholar
  32. Kusumi K, Komori H, Satoh H, Iba K (2000) Characterization of a zebra mutant of rice with increased susceptibility to light stress. Plant Cell Physiol 41:158–164PubMedGoogle Scholar
  33. Leon-Kloosterziel KM, Gil MA, Ruijs GJ et al (1996) Isolation and characterization of abscisic acid-deficient Arabidopsis mutants at two new loci. Plant J 10:655–661CrossRefPubMedGoogle Scholar
  34. Li F, Murillo C, Wurtzel E (2007) Maize Y9 encodes a product essential for 15-cis-zeta-carotene isomerization. Plant Physiol 144:1181–1189CrossRefPubMedGoogle Scholar
  35. Liu XQ, Bai XQ, Wang XJ, Chu CC (2007) OsWRKY71, a rice transcription factor, is involved in rice defense response. J Plant Physiol 164:969–979CrossRefPubMedGoogle Scholar
  36. Luo A, Qian Q, Yin H et al (2006) EUI1, encoding a putative cytochrome P450 monooxygenase, regulates internode elongation by modulating gibberellin responses in rice. Plant Cell Physiol 47:181–191CrossRefPubMedGoogle Scholar
  37. Masamoto K, Wada H, Kaneko T, Takaichi S (2001) Identification of a gene required for cis-to-trans carotene isomerization in carotenogenesis of the cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 42:1398–1402CrossRefPubMedGoogle Scholar
  38. Masamoto K, Hisatomi S, Sakurai I, Gombos Z, Wada H (2004) Requirement of carotene isomerization for the assembly of photosystem II in Synechocystis sp. PCC 6803. Plant Cell Physiol 45:1325–1329CrossRefPubMedGoogle Scholar
  39. Naested H, Holm A, Jenkins T et al (2004) Arabidopsis VARIEGATED 3 encodes a chloroplast-targeted, zinc-finger protein required for chloroplast and palisade cell development. J Cell Sci 117:4807–4818CrossRefPubMedGoogle Scholar
  40. Niyogi KK (1999) Photoprotection revisited: genetic and molecular approaches. Annu Rev Plant Physiol Plant Mol Biol 50:333–359CrossRefPubMedGoogle Scholar
  41. Oki S, Gu X, Kofoid KD, Liang GH (1997) A light-intensity sensitive chlorophyll mutant in sorghum. Hereditas 126:239–245CrossRefGoogle Scholar
  42. Park H, Kreunen SS, Cuttriss AJ, DellaPenna D, Pogson BJ (2002) Identification of the carotenoid isomerase provides insight into carotenoid biosynthesis, prolamellar body formation, and photomorphogenesis. Plant Cell 14:321–332CrossRefPubMedGoogle Scholar
  43. Pogson BJ, Niyogi KK, Bjorkman O, DellaPenna D (1998) Altered xanthophyll compositions adversely affect chlorophyll accumulation and nonphotochemical quenching in Arabidopsis mutants. Proc Natl Acad Sci USA 95:13324–13329CrossRefPubMedGoogle Scholar
  44. Qin G, Gu H, Ma L, Peng Y, Deng XW, Chen Z, Qu LJ (2007) Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis. Cell Res 17:471–482CrossRefPubMedGoogle Scholar
  45. Rosso D, Ivanov AG, Fu A et al (2006) IMMUTANS does not act as a stress-induced safety valve in the protection of the photosynthetic apparatus of Arabidopsis during steady-state photosynthesis. Plant Physiol 142:574–585CrossRefPubMedGoogle Scholar
  46. Sakamoto W (2003) Leaf-variegated mutations and their responsible genes in Arabidopsis thaliana. Genes. Genet Syst 78:1–9CrossRefGoogle Scholar
  47. Takechi K, Sodmergen Murata M, Motoyoshi F, Sakamoto W (2000) The YELLOW VARIEGATED (VAR2) locus encodes a homologue of FtsH, an ATP-dependent protease in Arabidopsis. Plant Cell Physiol 41:1334–1346CrossRefPubMedGoogle Scholar
  48. Tong H, Jin Y, Liu W et al (2009) DWARF AND LOW-TILLERING, a new member of the GRAS family, plays positive roles in brassinosteroid signaling in rice. Plant J 58:803–816CrossRefPubMedGoogle Scholar
  49. Welsch R, Wüst F, Bär C, Al-Babili S, Beyer P (2008) A third phytoene synthase is devoted to abiotic stress-induced abscisic acid formation in rice and defines functional diversification of phytoene synthase genes. Plant Physiol 147:367–380CrossRefPubMedGoogle Scholar
  50. Werner B, Burton G (1991) Dominant zebra mutant in pearl millet. J Hered 82:68–69Google Scholar
  51. Wetzel CM, Jiang CZ, Meehan LJ, Voytas DF, Rodermel SR (1994) Nuclear-organelle interactions: the immutans variegation mutant of Arabidopsis is plastid autonomous and impaired in carotenoid biosynthesis. Plant J 6:161–175CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Chenglin Chai
    • 1
  • Jun Fang
    • 1
  • Yang Liu
    • 1
    • 2
  • Hongning Tong
    • 1
  • Yanqing Gong
    • 1
  • Yiqin Wang
    • 1
  • Min Liu
    • 1
  • Youping Wang
    • 2
  • Qian Qian
    • 3
  • Zhukuan Cheng
    • 1
  • Chengcai Chu
    • 1
  1. 1.State Key Laboratory of Plant Genomics and National Centre for Plant Gene Research, Institute of Genetics and Developmental BiologyChinese Academy of SciencesChaoyang DistrictChina
  2. 2.Yangzhou UniversityYangzhouChina
  3. 3.China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhouChina

Personalised recommendations