Skip to main content
SpringerLink
Log in

Decreased TK activity alters growth, yield and tolerance to low temperature and low light intensity in transgenic cucumber plants

  • Original Paper
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Key message

Four CsTK antisense transgenic cucumber plants were obtained. Decreased TK activity decreased the photosynthetic rate, seed germination rate, growth yield, and the tolerance to low temperature and weak light stress.

Abstract

Transketolase (TK, EC 2.2.1.1) is a key enzyme in the photosynthetic carbon reduction cycle (Calvin cycle). A cDNA fragment (526 bp) encoding transketolase was cloned from cucumber plants (Cucumis sativa L. cv ‘Jinyou 3’) by RT-PCR. The antisense expression [(PBI-CsTK(−)] vector containing the CsTK gene fragment was constructed. The resulting plasmid was introduced into the cucumber inbred lines ‘08-1’ using the agrobacterium-mediated method, and four antisense transgenic cucumber plants were obtained. Decreased CsTK expression either unaltered or slightly increased the mRNA abundance and activities of the other main enzymes in the Calvin cycle, however, it decreased the TK activity and net photosynthetic rate (Pn) in antisense transgenic cucumber leaves. Antisense plants showed decreases in the growth, ratio of female flowers and yield compared with the wild-type (WT) plants. The decrease in Pn, stomatal conductance (Gs), transpiration rate (Tr), photochemical efficiency (Fv/Fm) and actual photochemical efficiency of PSII (ΦPSII) and the increase in electrolyte leakage (EL) were greater in antisense transgenic plants than in WT plants under low temperature (5 °C) and low light intensity (100 μmol m−2 s−1).

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
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

TK:

Transketolase

GAPDH:

Glyceraldehyde-phosphate dehydrogenase

FBPase:

Fructose-1,6-bisphosphatase

FBA:

Fructose-1,6-bisphosphate aldolase

RuBP:

Ribulose-1,5-bisphosphate

Fv/Fm:

Maximal photochemical effciency of PSII

SDS-PAGE:

Sodium dodecyl sulfate poly acrylamide gel electrophoresis

Ci:

Intercellular CO2 concentration

Gs:

Stomatal conductance

References

  • Bi HG, Wang ML, Jiang ZS, Dong XB, Ai XZ (2011) Impacts of suboptimal temperature and low light intensity on the activities and gene expression of photosynthetic enzymes in cucumber seedling leaves. Chin J Appl Ecol 22:2894–2900

    CAS  Google Scholar 

  • Bi HG, Wang ML, Dong XB, Ai XZ (2013) Cloning and expression analysis of transketolase gene in Cucumis sativus L. Plant Physiol Biochem 70:512–521

    Article  CAS  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  CAS  PubMed  Google Scholar 

  • Chen JH, Gibson JL, McCueg LA, Tabita FR (1991) Identification, expression, and deduced primary structure of transketolase and other enzymes encoded within the form II CO2 fixation operon of Rhodobacter sphaeroides. J Biol Chem 266:20447–20452

    CAS  PubMed  Google Scholar 

  • Flechner A, Dressen U, Westhoff P, Henze K, Schnarrenberger C, Martin W (1996) Molecular characterization of transketolase (EC 2.2. 1.1) active in the Calvin cycle of spinach chloroplasts. Plant Mol Biol 32:475–484

    Article  CAS  PubMed  Google Scholar 

  • Frank RAW, Leeper FJ, Luisi BF (2007) Structure, mechanism and catalytic duality of thiamine-dependent enzymes. Cell Mol Life Sci 64:892–905

    Article  CAS  PubMed  Google Scholar 

  • Haake VH, 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  CAS  PubMed  Google Scholar 

  • Haake V, Geiger M, Walch-Liu P, Engels C, Zrenner R, Stitt M (1999) Changes in aldolase activity in wild-type potato plants are important for acclimation to growth irradiance and carbon dioxide concentration, because plastid aldolase exerts control over the ambient rate of photosynthesis across a range of growth conditions. Plant J 17:479–489

    Article  CAS  Google Scholar 

  • Henkes S, Sonnewald U, Badur R, Flachmann R, Stitt M (2001) A small decrease of plastid transketolase activity in antisense tobacco transformants has dramatic effects on photosynthesis and phenylpropanoid metabolism. Plant Cell 13:535–551

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hudson GS, Evans JR, von Caemmerer S, Arvidsson YBC, Andrews TJ (1992) Reduction of ribulose-1, 5-bisphosphate carboxylase/oxygenase content by antisense RNA reduces photosynthesis in transgenic tobacco plants. Plant Physiol 98:294–302

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jordan F (2003) Current mechanistic understanding of thiamin diphosphate-dependent enzymatic reactions. Nat Prod Rep 20:184–201

    Article  CAS  PubMed  Google Scholar 

  • Kluger R (1987) Thiamin diphosphate: a mechanistic update on enzymic and nonenzymic catalysis of decarboxylation. Chem Rev 87:863–876

    Article  CAS  Google Scholar 

  • Kochetov G (2001) Functional flexibility of the transketolase molecule. Biochem (Moscow) 66:1077–1085

    Article  CAS  Google Scholar 

  • Kovina MV, Kochetov GA (1998) Cooperativity and flexibility of active sites in homodimeric transketolase. FEBS Lett 440:81–84

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Metzger MH, Hollenberg CP (1994) Isolation and characterization of the Pichia stipitis transketolase gene and expression in a xylose-utilising Saccharomyces cerevisiae transformant. Appl Microbiol Biotechnol 42:319–325

    CAS  PubMed  Google Scholar 

  • Portis AR Jr, Parry MAJ (2007) Discoveries in Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase): a historical perspective. Photosynth Res 94:121–143

    Article  CAS  PubMed  Google Scholar 

  • Quick WP, Schurr U, Scheibe R, Schulze ED, Rodermel SR, Bogorad L, Stitt M (1991) Decreased ribulose-1,5-bisphosphate carboxylase-oxygenase in transgenic tobacco transformed with “antisense” rbcS: I. Impact on photosynthesis in ambient growth conditions. Planta 183:542–554

    Article  CAS  PubMed  Google Scholar 

  • Raines CA (2011) Increasing photosynthetic carbon assimilation in C3 plants to improve crop yield: current and future strategies. Plant Physiol 155:36–42

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Rao M, Terry N (1989) Leaf phosphate status, photosynthesis, and carbon partitioning in sugar beet: I. Changes in growth, gas exchange, and Calvin cycle enzymes. Plant Physiol 90:814–819

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schenk G, Duggleby RG, Nixon PF (1998) Properties and functions of the thiamin diphosphate dependent enzyme transketolase. Int J Biochem Cell Biol 30:1297–1318

    Article  CAS  PubMed  Google Scholar 

  • Schneider G, Lindqvist Y (1998) Crystallography and mutagenesis of transketolase: mechanistic implications for enzymatic thiamin catalysis. Biochim Biophys Acta 1385:387–398

    Article  CAS  PubMed  Google Scholar 

  • Sheng X, Liu YJ, Liu CB (2013) Theoretical studies on the common catalytic mechanism of transketolase by using simplified models. J Mol Graph Model 39:23–28

    Article  CAS  PubMed  Google Scholar 

  • Stitt MJ, Lunn Usadel B (2010) Arabidopsis and primary photosynthetic metabolism—more than the icing on the cake. Plant J 61(6):1067–1091

    Article  CAS  PubMed  Google Scholar 

  • Sui XL, Zhang BX, Zhang ZX, Mao SL, Wang LH (2005) Differences of photosynthetic characteristics and low light-tolerance in seedlings of four pepper cultivars. Acta Hortic Sin 32:222–227 (in Chinese)

    CAS  Google Scholar 

  • Veitch NJ, Mauger DA, Cazzulo JJ, Lindqvist Y, Barrett MP (2004) Transketolase from Leishmania mexicana has a dual subcellular localization. Biochem J 382:759–767

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Villafranca JJ, Axelrod B (1971) Heptulose synthesis from nonphosphorylated aldoses and ketoses by spinach transketolase. J Biol Chem 246:3126–3131

    CAS  PubMed  Google Scholar 

  • Wang YR, Chen LM, Pan JS, He HL, Cai R (2006) Establishment of high effective regeneration system in cucumber (Cucumis Sativus L.) and agrobacterium tumefaciens mediated genetic transformation. J Shanghai Jiaotong Univ (Agricultural Science) 2:152–155

    Google Scholar 

  • Zhao SJ, Li DQ (1999) Manual of the modern plant physiology. Science Press, Beijing

    Google Scholar 

  • Zhao J, Wang H, Pan JS, Cai R, Wu AZ (2004) In vitro culture and plantlet regeneration from cotyledonary nodes of cucumber (Cucumis sativus L.). J Shanghai Jiaotong Univ (Agricultural Science) 22:43–47

    Google Scholar 

Download references

Acknowledgments

This work was supported by National Plan of Key Basic Research in China (Contract No. 2009CB119000).

Conflict of interest

The authors declare that there is no conflict of interest.

Author information

Authors and Affiliations

  1. State Key Laboratory of Crop Biology, College of Horticulture Science and Engineering, Shandong Agricultural University, 61 Daizong St., Tai’an, 271018, Shandong, People’s Republic of China

    Huangai Bi, Xubing Dong, Guoxiu Wu, Meiling Wang & Xizhen Ai

Authors
  1. Huangai Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xubing Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guoxiu Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Meiling Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xizhen Ai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xizhen Ai.

Additional information

Communicated by Jim Register.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bi, H., Dong, X., Wu, G. et al. Decreased TK activity alters growth, yield and tolerance to low temperature and low light intensity in transgenic cucumber plants. Plant Cell Rep 34, 345–354 (2015). https://doi.org/10.1007/s00299-014-1713-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-014-1713-5

Keywords

Search

Navigation