Skip to main content
Springer Nature Link
Log in

Antisense suppression of violaxanthin de-epoxidase in tobacco does not affect plant performance in controlled growth conditions

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Violaxanthin de-epoxidase (VDE) catalyzes the de-epoxidation of violaxanthin to antheraxanthin and zeaxanthin in the xanthophyll cycle. Tobacco was transformed with an antisense VDE construct under control of the cauliflower mosaic virus 35S promoter to determine the effect of reduced levels of VDE on plant growth. Screening of 40 independent transformants revealed 18 antisense lines with reduced levels of VDE activity with two in particular (TAS32 and TAS39) having greater than 95% reduction in VDE activity. Northern analysis demonstrated that these transformants had greatly suppressed levels of VDE mRNA. De-epoxidation of violaxanthin was inhibited to such an extent that no zeaxanthin and only very low levels of antheraxanthin could be detected after exposure of leaves to high light (2000 μmol m−2 s−1 for 20 min) with no observable effect on levels of other carotenoids and chlorophyll. Non-photochemical quenching was greatly reduced in the antisense VDE tobacco, demonstrating that a significant level of the non-photochemical quenching in tobacco requires de-epoxidation of violaxanthin. Although the antisense plants demonstrated a greatly impaired de-epoxidation of violaxanthin, no effect on plant growth or photosynthetic rate was found when plants were grown at a photon flux density of 500 or 1000 μmol m−2 s−1 under controlled growth conditions as compared to wild-type tobacco.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Arvidsson P-O, Bratt CE, Carlsson M and Åkerlund H-E (1996) Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein. Photosynth Res 49: 119–129

    Article  CAS  Google Scholar 

  • Bugos RC and Yamamoto HY (1996) Molecular cloning of violaxanthin de-epoxidase from romaine lettuce and expression in Escherichia coli. Proc Natl Acad Sci USA 93: 6320–6325

    Article  PubMed  CAS  Google Scholar 

  • Bugos RC, Chang S-H and Yamamoto HY (1999) Developmental expression of violaxanthin de-epoxidase in leaves of tobacco growing under high and low light. Plant Physiol 121: 207–213

    Article  PubMed  CAS  Google Scholar 

  • Bugos RC, Hieber AD and Yamamoto HY (1998) Xanthophyll cycle enzymes are members of the lipocalin family, the first identified from plants. J Biol Chem 273: 15321–15324

    Article  PubMed  CAS  Google Scholar 

  • Church GM and Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995

    Article  PubMed  CAS  Google Scholar 

  • Demmig-Adams B and Adams WW III (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1: 21–26

    Article  Google Scholar 

  • Demmig-Adams B, Gilmore AM and Adams WW III (1996) In vivo functions of carotenoids in higher plants. FASEB J 10: 403–412

    PubMed  CAS  Google Scholar 

  • Doyle JJ and Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19: 11–15

    Google Scholar 

  • Eskling M, Arvidsson P-O and Åkerlund H-E (1997) The xanthophyll cycle, its regulation and components. Physiol Plant 100: 806–816

    Article  CAS  Google Scholar 

  • Feinberg AP and Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13

    Article  PubMed  CAS  Google Scholar 

  • Fisher DK and Guiltinan MJ (1995) Rapid, efficient production of homozygous transgenic tobacco plants with Agrobacterium tumefaciens: A seed-to-seed protocol. Plant Mol Biol Rep 13: 278–289

    CAS  Google Scholar 

  • Fourney RM, Miyakoshi J, Day RS III and Paterson MC (1988) Northern blotting: Efficient RNA staining and transfer. Focus 10: 5–7

    Google Scholar 

  • Gilmore AM (1997) Mechanistic aspects of xanthophyll cycledependent photoprotection in higher plant chloroplasts and leaves. Physiol Plant 99: 197–209

    Article  CAS  Google Scholar 

  • Gilmore AM and Yamamoto HY (1991) Resolution of lutein and zeaxanthin using a non-endcapped, lightly carbon-loaded C18 high-performance liquid chromatographic column. J Chromatogr 543: 137–145

    Article  CAS  Google Scholar 

  • Hager A (1969) Lichtbedingte pH-Erniedrigung in einem Chloroplasten-Kompartiment als Ursache der enzymatischen Violaxanthin-? Zeaxanthin-Umwandlung; Beziehungen zur Photophosphorylierung. Planta 89: 224–243

    Article  CAS  Google Scholar 

  • Hager A (1975) Die reversiblen, lichtabhängigen Xanthophyllumwandlungen im Chloroplasten. Ber Dtsch Bot Ges Bd 88: 27–44

    CAS  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG and Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227: 1229–1231

    CAS  Google Scholar 

  • Murashige T and Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497

    Article  CAS  Google Scholar 

  • Niyogi KK, Björkman O and Grossman AR(1997a) Chlamydomonas xanthophyll cycle mutants identified by video imaging of chlorophyll fluorescence quenching. Plant Cell 9: 1369–1380

    Article  PubMed  CAS  Google Scholar 

  • Niyogi KK, Björkman O and Grossman AR (1997b) The roles of specific xanthophylls in photoprotection. Proc Natl Acad Sci USA 94: 14162–14167

    Article  PubMed  CAS  Google Scholar 

  • Niyogi KK, Grossman AR and Björkman O (1998) Arabidopsis mutants define a central role for the xanthophyll cycle in the regulation of photosynthetic energy conversion. Plant Cell 10: 1121–1134

    Article  PubMed  CAS  Google Scholar 

  • Rockholm DC and Yamamoto HY (1993) Purification of violaxanthin de-epoxidase by lipid affinity precipitation. In: Yamamoto HY and Smith CM (eds) Photosynthetic Responses to the Environment, p 237. American Society of Plant Physiologists, Rockville, Maryland

    Google Scholar 

  • Rockholm DC and Yamamoto HY (1996) Violaxanthin deepoxidase: Purification of a 43-kilodalton lumenal protein from lettuce by lipid-affinity precipitation with monogalactosyldiacylglyceride. Plant Physiol 110: 697–703

    Article  PubMed  CAS  Google Scholar 

  • Schreiber U, Schliwa U and Bilger W (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer. Photosynth Res 10: 51–62

    Article  CAS  Google Scholar 

  • Siefermann D and Yamamoto HY (1975) Properties of NADPH and oxygen-dependent zeaxanthin epoxidation in isolated chloroplasts. A transmembrane model for the violaxanthin cycle. Arch Biochem Biophys 171: 70–77

    Article  PubMed  CAS  Google Scholar 

  • Stürzl M and Roth WK (1990) 'Run-Off' synthesis and application of defined single-stranded DNA hybridization probes. Anal Biochem 185: 164–169

    Article  PubMed  Google Scholar 

  • Theologis A, Oeller PW, Wong L-M, Rottmann WH and Gantz DM (1993) Use of a tomato mutant constructed with reverse genetics to study fruit ripening, a complex developmental process. Dev Genet 14: 282–295

    Article  PubMed  CAS  Google Scholar 

  • Van Kooten O and Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 25: 147–150

    Article  Google Scholar 

  • Yamamoto HY (1985) Xanthophyll cycles. Methods Enzymol 110: 303–312

    Article  CAS  Google Scholar 

  • Yamamoto HY and Higashi RM (1978) Violaxanthin de-epoxidase. Lipid composition and substrate specificity. Arch Biochem Biophys 190: 514–522

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto HY and Kamite L (1972) The effects of dithiothreitol on violaxanthin de-epoxidation and absorbance changes in the 500 nm region. Biochim Biophys Acta 267: 538–543

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto HY, Nakayama TOM and Chichester CO (1962) Studies on the light and dark interconversions of leaf xanthophylls. Arch Biochem Biophys 97: 168–173

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto HY, Kamite L and Wang Y-Y (1972) An ascorbateinduced absorbance change in chloroplasts from violaxanthin de-epoxidation. Plant Physiol 49: 224–228

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biosciences and Biosystems Engineering, University of Hawaii-Manoa, 1955 East West Road, Room 218, Honolulu, HI, 96822, USA

    Sue-Hwei Chang, Robert C. Bugos & Wen-Hao Sun

  2. Department of Molecular Biosciences and Biosystems Engineering, University of Hawaii-Manoa, 1955 East West Road, Room 218, Honolulu, HI, 96822, USA

    Harry Y. Yamamoto

Authors
  1. Sue-Hwei Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert C. Bugos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wen-Hao Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Harry Y. Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Harry Y. Yamamoto.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chang, SH., Bugos, R.C., Sun, WH. et al. Antisense suppression of violaxanthin de-epoxidase in tobacco does not affect plant performance in controlled growth conditions. Photosynthesis Research 64, 95–103 (2000). https://doi.org/10.1023/A:1026518524426

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1026518524426

Search

Navigation