Skip to main content
SpringerLink
Log in

Genetic engineering of the biosynthesis of glycinebetaine leads to increased tolerance of photosynthesis to salt stress in transgenic tobacco plants

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Genetically engineered tobacco (Nicotiana tabacum L.) with the ability to synthesis glycinebetaine (GB) in chloroplasts was established by introducing the BADH gene for betaine aldehyde dehydrogenase from spinach (Spinacia oleracea L.). The genetic engineering resulted in enhanced tolerance of growth of young seedlings to salt stress. This increased tolerance was not due to improved water status, since there were no significant differences in accumulation of sodium and chloride, leaf water potential, and relative water content between wild type and transgenic plants under salt stress. Salt stress resulted in a decrease in CO2 assimilation and such a decrease was much greater in wild type plants than in transgenic plants. Though salt stress showed no damage to PSII, there were a decrease in the maximal PSII electron transport rate in vivo and an increase in non-photochemical quenching (NPQ) and these changes were greater in wild type plants than in transgenic plants. In addition, salt stress inhibited the activities of ribulose 1,5-bisphosphate carboxylase/oxygenase, chloroplastic fructose-1,6-bisphosphatase, fructose-1,6-bisphosphate aldolase, and phosphoribulokinase and such a decrease was also greater in wild type plants than in transgenic plants, suggesting that GB protects these enzymes against salt stress. However, there were no significant changes in the activities of phosphoglycerate kinase, triose phosphate isomerase, ribulose-5-phosphate isomerase, transketolase, and sedoheptulose-1,7-bisphosphatase in both wild type and transgenic plants. The results in this study suggest that enhanced tolerance of CO2 assimilation to salt stress may be one of physiological bases for increased tolerance of growth of transgenic plants to salt stress.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abadía A, Belkohodja R, Morales F, Abadía J (1999) Effects of salinity on the photosynthetic pigment composition of barley (Hordeum vulgare L.) growth under a triple-line-source sprinkler system in the field. J Plant Physiol 154:392–400

    Google Scholar 

  • Alia, Hayashi H, Chen THH, Murata N (1998a) Transformation with a gene for choline oxidase enhances the cold tolerance of Arabidopsis during germination and early growth. Plant Cell Environ 21:232–239

    Article  CAS  Google Scholar 

  • Alia, Hayashi H, Sakamoto A, Murata N (1998b) Enhancement of the tolerance of Arabidopsis to high temperatures by genetic engineering of the synthesis of glycinebetaine. Plant J 16:155–161

    Article  PubMed  CAS  Google Scholar 

  • Andrews TJ, Hudson GS, Mate CJ, von Caemmerer S, Evans JR, Avridsson YBC (1995) Rubisco, consequences of altering its expression and activation in transgenic plants. J Exp Bot 46:1293–1300

    Article  CAS  Google Scholar 

  • Belkhodja R, Morales F, Abadia A, Gomez-Aparisi J, Abadia J (1994) Chlorophyll fluorescence as a possible tool for salinity tolerance screening in barley (Hordeum vulgare L.). Plant Physiol 104:667–673

    PubMed  CAS  Google Scholar 

  • Bourot S, Sire O, Trautwetter A, Touze T, Wu LF, Blanco C, Bernard T (2000) Glycinebetaine-assisted protein folding in a lysA mutant of Escherichia coli. J Biol Chem 275:1050–1056

    Article  PubMed  CAS  Google Scholar 

  • Boyer JS (1982) Plant productivity and environment. Science 218:443–448

    Article  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein using the principal of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Brugnoli E, Björkman O (1992) Growth of cotton under continuous salinity stress: influence on allocation pattern, stomatal and non-stomatal components of photosynthesis and dissipation of excess light energy. Planta 187:335–345

    Article  CAS  Google Scholar 

  • Downton WJ, Grant WJR, Robinson SP (1985) Photosynthetic and stomatal response of spinach leaves to salt stress. Plant Physiol 77:85–88

    Article  Google Scholar 

  • Eckhardt NA, Portis AR Jr (1997) Heat denaturation profiles of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and Rubisco activase and the inability of Rubisco activase to restore activity of heat-denaturated Rubisco. Plant Physiol 113:243–248

    Google Scholar 

  • Epstein E, Norlyn JD, Rush DW, Kingsbury RW, Kelly DB, Cunningham GA, Wrona AF (1980) Saline culture of crops: a genetic approach. Science 210:399–404

    Article  PubMed  CAS  Google Scholar 

  • Everard JD, Gucci R, Kann SC, Flore JA, Loescher WH (1994) Gas exchange and carbon partitioning in the leaves of celery (Apium graveolens L.) at various levels of root zone salinity. Plant Physiol 106:281–292

    PubMed  CAS  Google Scholar 

  • Feller U, Crafts-Brandner SJ, Salvucci E (1998) Moderately high temperatures inhibit ribulose-1,5-bisphosphate carboxylase/oxygenase activase-mediated activation of Rubisco. Plant Physiol 116:539–546

    Article  PubMed  CAS  Google Scholar 

  • Flower TJ, Troke PF, Yeo AR (1977) The mechanism of salt tolerance in halophytes. Annu Rev Plant Physiol 28:89–121

    Article  Google Scholar 

  • Genty B, Briantais JM, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 99:87–92

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Gorham J (1995) Betaines in higher plants-biosynthesis and role in stress metabolism. In: Wallsgrove RM (ed) Amino acids and their derivatives in higher plants. Cambridge University Press, Cambridge, pp 171–203

    Google Scholar 

  • Haake V, Zrenner R, Sonnewald U, Stitt M (1998) A moderate decrease of plastid aldolase activity inhibits photosynthesis, alters the levels of sugars and starch, and inhibits growth of potato plants. Plant J 14:147–157

    Article  PubMed  CAS  Google Scholar 

  • Hayashi H, Alia, Mustardy L, Deshnium P, Ida M, Murata N (1997) Transformation of Arabidopsis thaliana with the coda gene for choline oxidase: accumulation of glycinebetaine and enhanced tolerance to salt and cold stress. Plant J 12:133–142

    Article  PubMed  CAS  Google Scholar 

  • Holmström K-O, Somersalo S, Mandal A, Palva ET, Welin B (2000) Improved tolerance to salinity and low temperature in transgenic tobacco producing glycine betaine. J Exp Bot 51:177–185

    Article  PubMed  Google Scholar 

  • Horton P, Ruban AV, Walters RG (1996) Regulation of light harvesting in green plants. Annu Rev Plant Physiol Plant Mol Biol 47:655–684

    Article  PubMed  CAS  Google Scholar 

  • Incharoensakdi A, Takabe T, Akazawa T (1986) Effect of betaine on enzyme activity and subunit interaction of ribulose 1,5-bisphosphate carboxylase/oxygenase from Aphanothece halophytica. Plant Physiol 81:1044–1049

    PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Leegood RC (1990) Enzymes of the Calvin cycle. In: Lea PJ (ed) Methods in plant biochemistry, vol 3. Academic Press, London, pp 16–45

    Google Scholar 

  • Long SP, Baker NR (1986) Saline terrestrial environments. In: Baker NR, Long SP (eds) Photosynthesis in contrasting environments. Elsevier, Amsterdam, pp 63–102

    Google Scholar 

  • Lu C, Zhang J (1998) Thermostability of photosystem II is increased in salt-stressed sorghum. Aust J Plant Physiol 25:317–324

    Article  CAS  Google Scholar 

  • Lu C, Qiu N, Lu Q, Wang B, Kuang T (2002) Does salt stress lead to increased susceptibility of photosystem II to photoinhibition and changes in photosynthetic pigment composition in halophyte Suaeda salsa grown outdoors? Plant Sci 163:1063–1068

    Article  CAS  Google Scholar 

  • Lu C, Jiang G, Wang B, Kuang T. (2003a) Photosystem II photochemistry and photosynthetic pigment composition in salt-adapted halophyte Artimisia anethifolia grown under outdoor conditions. J Plant Physiol 160:403–408

    Article  PubMed  CAS  Google Scholar 

  • Lu C, Qiu N, Wang B, Zhang J (2003b) Salinity treatment shows no effects on photosystem II photochemistry but increases the resistance of photosystem II to heat stress in halophyte Suaeda salsa. J Exp Bot 54:851–860

    Article  PubMed  CAS  Google Scholar 

  • Mishra SK, Subrahmanyam D, Singhal GS (1991) Interactionship between salt and light stress on the primary process of photosynthesis. J Plant Physiol 138:92–96

    CAS  Google Scholar 

  • Morales F, Abadia A, Gomez-Aparis J, Abadia J (1992) Effects of combined NaCl and CaCl2 salinity on photosynthetic parameters of barley grown in nutrient solution. Physiol Plant 86:419–426

    Article  CAS  Google Scholar 

  • Mullet JE, Chua NH (1983) In vitro reconstitution of synthesis, uptake, and assembly of cytoplasmically synthesized chloroplast proteins. Meth Enzymol 97:502–509

    Article  CAS  Google Scholar 

  • Nomura M, Hibino T, Takabe T, Sugyama T, Yokota A, Miyake H, Takabe T (1998) Transgenically produced glycinebetaine protects ribulose 1,5-bisphosphate carboxylase/oxygenase from inactivation in Synechococcus sp. PCC7942 under salt stress. Plant Cell Physiol 39:425–432

    CAS  Google Scholar 

  • Papageorgiou GC, Murata N (1995) The unusually strong stabilizing effects of glycine betaine on the structure and function of the oxygen-evolving photosystem II complex. Photosynth Res 44:243–252

    Article  CAS  Google Scholar 

  • Park E-J, Jeknic´ Z, Sakamoto A, DeNomal J, Yuwansiri R, Murata N, Chen THH (2004) Genetic engineering of glycinebetaine synthesis in tomato protects seeds, plants, and flowers from chilling damage. Plant J 40:474–487

    Article  PubMed  CAS  Google Scholar 

  • Perchorowicz JT, Raynes DA, Jensen RG (1981) Light limitation of photosynthesis and activation of ribulose bisphosphate carboxylase in wheat seedlings. Proc Natl Acad Sci USA 78:2985–2989

    Article  PubMed  CAS  Google Scholar 

  • Pollard A, Wyn Jones RG (1979) Enzyme activities in concentrated solutions of glycinebetaine and other solutes. Planta 144:291–298

    Article  CAS  Google Scholar 

  • Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophyll 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 

  • Rhodes D, Hanson AD (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants. Annu Rev Plant Physiol Plant Mol Biol 44:357–384

    Article  CAS  Google Scholar 

  • Sakamoto A, Alia, Murata N (1998) Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold. Plant Mol Biol 38:1011–1019

    Article  PubMed  CAS  Google Scholar 

  • Sakamoto A, Murata N (2001) The use of bacterial choline oxidase, a glycine betaine-synthesizing enzyme, to create stress-resistant transgenic plants. Plant Physiol 125:180–188

    Article  PubMed  CAS  Google Scholar 

  • Sakamoto A, Murata N (2002) The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant Cell Environ 25:163–171

    Article  PubMed  CAS  Google Scholar 

  • Sakamoto A, Valverde R, Alia, Chen THH, Murata N (2000) Transformation of Arabidopsis with the coda gene for choline oxidase enhances freezing tolerance of plants. Plant J 22:449–453

    Article  PubMed  CAS  Google Scholar 

  • Saneoka H, Nagasaka C, Hahn DT, Yang WJ, Premachandra GS, Joly RJ, Rhodes D (1995) Salt tolerance of glycinebetaine-deficient and containing maize lines. Plant Physiol 107:631–638

    PubMed  CAS  Google Scholar 

  • Schimakat D, Heineke D, Heldt HW (1990) Regulation of sedoheptulose-1,7-bisphosphatase by sedohepulose-7-phosphate and glycerate, and fructose-1,6-bisphosphatase by glycerate in spinach chloroplasts. Planta 181:97–103

    Google Scholar 

  • Schrader SM, Wise RR, Wacholtz WE, Ort DR, Sharkey TD (2004) Thylakoid membrane responses to moderately high leaf temperature in Pima cotton. Plant Cell Environ 27:725–735

    Article  CAS  Google Scholar 

  • Su J, Hirji R, Zhang L, He C., Selvaraj G., Wu R (2006) Evaluation of the stress-inducible production of choline oxidase in transgenic rice as a strategy for producing the stress-protectant glycine betaine. J Exp Bot 57:1129–1135

    Article  PubMed  CAS  Google Scholar 

  • Stitt M (1986) Limitation of photosynthesis by carbon metabolism. I. Evidence for excess electron transport capacity in leaves carrying out photosynthesis in saturating light and CO2. Plant Physiol 81:1115–1122

    PubMed  CAS  Google Scholar 

  • Szabolcs L (1989) Salt-affected soils. CRC Press, Inc., Boca Raton

    Google Scholar 

  • Tanji KK (1990) Nature and extent of agricultural salinity. In: Tanji KK (ed) Agricultural salinity assessment and management. American Society of Civil Engineers, New York, pp 1–17

    Google Scholar 

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

    Article  Google Scholar 

  • Yang X, Liang Z, Lu C (2005) Genetic engineering of the biosynthesis of glycinebetaine enhances photosynthesis against high temperature stress in transgenic tobacco plants. Plant Physiol 138:2299–2309

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Frontier Project of the Knowledge Innovation Engineering of the Chinese Academy of Sciences (grants number: KSCX2-YW-N-042) and the Program of 100 Distinguished Young Scientists of Chinese Academy of Sciences to C. Lu.

Author information

Authors and Affiliations

  1. Photosynthesis Research Center, Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, China

    Xinghong Yang, Zheng Liang, Xiaogang Wen & Congming Lu

  2. College of Life Science, Key Laboratory of Crop Biology of Shandong Province, Shandong Agricultural University, Taian, 211018, China

    Xinghong Yang

  3. National Plant Gene Center, Beijing, 100093, China

    Congming Lu

Authors
  1. Xinghong Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zheng Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaogang Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Congming Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Congming Lu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, X., Liang, Z., Wen, X. et al. Genetic engineering of the biosynthesis of glycinebetaine leads to increased tolerance of photosynthesis to salt stress in transgenic tobacco plants. Plant Mol Biol 66, 73–86 (2008). https://doi.org/10.1007/s11103-007-9253-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11103-007-9253-9

Keywords

Search

Navigation