Skip to main content
SpringerLink
Log in

Isolation of a citrus promoter specific for reproductive organs and its functional analysis in isolated juice sacs and tomato

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

Abstract

While searching for genes expressed in acid lemon but not in acidless lime pulp, we isolated clone Cl111 which showed the following expression phenotypes: (1) while it was expressed in the ovaries in both varieties, its mRNA was detected only in the pulp of the acid fruit, (2) no or very low expression of the gene was detected in vegetative organs. These expression patterns suggested that Cl111 is an ovary- and pulp-specific gene. The ability of ~2-kb fragments upstream of the transcription start site of the lemon and lime genes to confer reporter-gene activity was investigated by transient expression in isolated juice vesicles of both varieties. Whereas Cl111 promoter from lemon showed faint activity in lemon and lime juice vesicles, no activity was evident with the lime promoter. The activities of the 2-kb fragments and their delimited fragments were further investigated in tomato. The results indicated that the promoters were active in a manner similar to that in acid lemon and acidless lime: the lemon promoter generated activity in the fruit endocarp, analogous to citrus fruit pulp. The delimitation analyses identified an expression-conferring region which, in the lemon promoter, contained a sequence homologous to a fruit-specific element of the melon cucumisin gene. Another region, which reduced promoter activity, contained an I-Box-like sequence, identified as a fruit-specific negative element. Taken together, Cl111 promoter was confirmed to be pulp- and flower-specific. Differences in the expression of Cl111 between the two varieties could be attributable to changes in the gene promoter region.

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

  • Agius F, Amaya I, Botella MA, Valpuesta V (2005) Functional analysis of homologous and heterologous promoters in strawberry fruits using transient expression. J Exper Bot 56:37–46

    CAS  Google Scholar 

  • Almeida DPF, Huber DJ (2001) Transient increase in locular pressure and occlusion of endocarpic apertures in ripening tomato fruit. J Plant Physiol 158:199–203

    Article  CAS  Google Scholar 

  • Atkinson RG, Bolitho KM, Wright MA, Iturriagagoitia-Bueno T, Reid SJ, Ross GS (1998) Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato. Plant Mol Biol 38:449–460

    Article  PubMed  CAS  Google Scholar 

  • Avsian-Kretchmer O, Gueta-Dahan Y, Lev-Yadun S, Gollop R, Ben-Hayyim G (2004) The salt-stress signal transduction pathway that activates the gpx1 promoter is mediated by intracellular H2O2, different from the pathway induced by extracellular H2O2. Plant Physiol 135:1685–1696

    Article  PubMed  CAS  Google Scholar 

  • Becker D, Kemper E, Schell J, Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left t-DNA border. Plant Mol Biol 20:1195–1197

    Article  PubMed  CAS  Google Scholar 

  • Carmi N, Salts Y, Dedicova B, Shabtai S, Barg R (2003) Induction of parthenocarpy in tomato via specific expression of the rolB gene in the ovary. Planta 217:726–735

    Article  PubMed  CAS  Google Scholar 

  • Cutter EG (1971) Plant Anatomy: experiment and interpatation. Part 2: Organs. Edward Arnold, London

    Google Scholar 

  • de Azevedo FA, Mourao FDA, Schinor EH, de Paoli LG, Mendes BMJ, Harakava R, Gabriel DW, Lee RF (2006) GUS gene expression driven by a citrus promoter in transgenic tobacco and ‘Valencia’ sweet orange. Pesq Agro Brasil 41:1623–1628

    Google Scholar 

  • Deikman J, Kline R, Fischer RL (1992) Organization of ripening and ethylene regulatory regions in a fruit-specific promoter from tomato (Lycopersicon-esculentum). Plant Physiology 100:2013–2017

    Article  PubMed  CAS  Google Scholar 

  • Dominguez O, Lopezlarrea C (1994) Gene walking by unpredictably primed pcr. Nucl Acids Res 22:3247–3248

    Article  PubMed  CAS  Google Scholar 

  • duJardin P, Harvengt L, Kirsch F, Le VQ, NguyenQuoc B, Yelle S (1997) Sink-cell-specific activity of a potato ADP-glucose pyrophosphorylase B-subunit promoter in transgenic potato and tomato plants. Planta 203:133–139

    Article  CAS  Google Scholar 

  • Endo T, Shimada T, Fujii H, Moriguchi T, Omura M (2007) Promoter analysis of a type 3 metallothionein-like gene abundant in Satsuma mandarin (Citrus unshiu Marc.) fruit. Sci Hort 112:207–214

    Article  CAS  Google Scholar 

  • Fahn A (1974) Plant anatomy. Pergamon Press, Oxford

    Google Scholar 

  • Fraser PD, Romer S, Shipton CA, Mills PB, Kiano JW, Misawa N, Drake RG, Schuch W, Bramley PM (2002) Evaluation of transgenic tomato plants expressing an additional phytoene synthase in a fruit-specific manner. Proc Natl Acad Sci USA 99:1092–1097

    Article  PubMed  CAS  Google Scholar 

  • Ho LC, Hewitt JD (1986) Fruit development. In: Atherton AJ, Rudich J (eds) The tomato crop. A scientific basis for improvement. Chapman and Hall, London, pp 201–240

    Google Scholar 

  • Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New agrobacterium helper plasmids for gene-transfer to plants. Transgenic Res 2:208–218

    Article  CAS  Google Scholar 

  • Jeon JS, Chung YY, Lee S, Yi GH, Oh BG, An GH (1999) Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.). Plant Mol Biol 39:35–44

    Article  PubMed  CAS  Google Scholar 

  • Karaaslan M, Hrazdina G (2010) Characterization of an expansin gene and its ripening-specific promoter fragments from sour cherry (Prunus cerasus L.) cultivars. Acta Physiol Plant 32:1073–1084

    Article  CAS  Google Scholar 

  • Kneissl ML, Deikman J (1996) The tomato E8 gene influences ethylene biosynthesis in fruit but not in flowers. Plant Physiol 112:537–547

    PubMed  CAS  Google Scholar 

  • Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB (1990) Different temporal and spatial gene-expression patterns occur during anther development. Plant Cell 2:1201–1224

    Article  PubMed  CAS  Google Scholar 

  • Kosugi S, Ohashi Y, Nakajima K, Arai Y (1990) An improved assay for beta-glucuronidase in transformed-cells—methanol almost completely suppresses a putative endogenous beta-glucuronidase activity. Plant Sci 70:133–140

    Article  CAS  Google Scholar 

  • Kuntz M, Chen HC, Simkin AJ, Romer S, Shipton CA, Drake R, Schuch W, Bramley PM (1998) Upregulation of two ripening-related genes from a nonclimacteric plant (pepper) in a transgenic climacteric plant (tomato). Plant J 13:351–361

    Article  CAS  Google Scholar 

  • Lau JM, Cooper NG, Robinson DL, Korban SS (2009) Sequence and in silico characterization of the tomato polygalacturonase (PG) promoter and terminator regions. Plant Mol Biol Rep 27:250–256

    Article  CAS  Google Scholar 

  • Lin JY, Fan R, Wan XR, Charng YY, Wang NN (2007) Structural analysis of the promoter of tomato 1-aminocyclopropane-1-carboxylate synthase 6 gene (Le-ACS6). Chin Sci Bull 52:1217–1222

    Article  CAS  Google Scholar 

  • McCormick S (1991) Transformation of tomato with Agrobacterium tumefaciens. Plant Tiss Cult Man B6:1–9

    Google Scholar 

  • Nain V, Verma A, Kumar N, Sharma P, Ramesh B, Kumar PA (2008) Cloning of an ovule specific promoter from Arabidopsis thaliana and expression of beta-glucuronidase. Indian J Experim Biol 46:207–211

    CAS  Google Scholar 

  • Omidvar V, Abdullah SNA, Izadfard A, Ho CL, Mahmood M (2010) The oil palm metallothionein promoter contains a novel AGTTAGG motif conferring its fruit-specific expression and is inducible by abiotic factors. Planta 232:925–936

    Article  PubMed  CAS  Google Scholar 

  • Potenza C, Aleman L, Sengupta-Gopalan C (2004) Targeting transgene expression in research, agricultural, and environmental applications: Promoters used in plant transformation. In Vitro Cell Dev Biol Plant 40:1–22

    CAS  Google Scholar 

  • Rasori A, Bertolasi B, Furini A, Bonghi C, Tonutti P, Ramina A (2003) Functional analysis of peach ACC oxidase promoters in transgenic tomato and in ripening peach fruit. Plant Sci 165:523–530

    Article  CAS  Google Scholar 

  • Rombauts S, Florquin K, Lescot M, Marchal K, Rouze P, Van de Peer Y (2003) Computational approaches to identify promoters and cis-regulatory elements in plant genomes. Plant Physiol 132:1162–1176

    Article  PubMed  CAS  Google Scholar 

  • Roth I (1977) Fruits of angiosperms. Gebruder Borntraeger, Berlin

    Google Scholar 

  • Sadka A, Dahan E, Cohen L, Marsh KB (2000) Aconitase activity and expression during the development of lemon fruit. Physiol Plant 108:255–262

    Article  CAS  Google Scholar 

  • Sadka A, Dahan E, Or E, Roose ML, Cohen L (2001) A comparative analysis of mitochondrial citrate synthase gene structure, transcript level and enzymatic activity in acidless and acid-containing Citrus varieties. Aust J Plant Physiol 28:383–390

    CAS  Google Scholar 

  • Schneider H (1968) The anatomy of citrus. In: Reuther R, Batchelor LD, Webber HJ (eds) The citrus industry, vol 2. Anatomy, physiology, genetics and reproduction. University of California, Riverside, pp 2–85

    Google Scholar 

  • Shlizerman L, Marsh K, Blumwald E, Sadka A (2007) Iron-shortage-induced increase in citric acid content and reduction of cytosolic aconitase activity in Citrus fruit vesicles and calli. Physiol Plant 131:72–79

    Article  PubMed  CAS  Google Scholar 

  • Twell D, Yamaguchi J, McCormick S (1990) Pollen-specific gene-expression in transgenic plants—coordinate regulation of 2 different tomato gene promoters during microsporogenesis. Development 109:705

    Google Scholar 

  • Vanhaaren MJJ, Houck CM (1991) Strong negative and positive regulatory elements contribute to the high-level fruit-specific expression of the tomato 2A11 gene. Plant Mol Biol 17:615–630

    Article  CAS  Google Scholar 

  • Vanhaaren MJJ, Houck CM (1993) A functional map of the fruit-specific promoter of the tomato 2A11-gene. Plant Mol Biol 21:625–640

    Article  CAS  Google Scholar 

  • Wakeley PR, Rogers HJ, Rozycka M, Greenland AJ, Hussey PJ (1998) A maize pectin methylesterase-like gene, ZmC5, specifically expressed in pollen. Plant Mol Biol 37:187–192

    Article  PubMed  CAS  Google Scholar 

  • Xu RL, Goldman S, Coupe S, Deikman J (1996) Ethylene control of E4 transcription during tomato fruit ripening involves two cooperative cis elements. Plant Mol Biol 31:1117–1127

    Article  PubMed  CAS  Google Scholar 

  • Yamagata H, Yonesu K, Hirata A, Aizono Y (2002) TGTCACA motif is a novel cis-regulatory enhancer element involved in fruit-specific expression of the cucumisin gene. J Biol Chem 277:11582–11590

    Article  PubMed  CAS  Google Scholar 

  • Yin T, Wu HY, Zhang SL, Liu JM, Lu HY, Zhang LX, Xu Y, Chen DM (2009) Two negative cis-regulatory regions involved in fruit-specific promoter activity from watermelon (Citrullus vulgaris S.). J Experim Bot 60:169–185

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by fund number 203-462 of the Chief Scientist of the Ministry of Agriculture and Rural Development. We would like to acknowledge Dr. Einat Sadot (The Volcani Center, ARO, Israel) for the donation of control plasmid (CaMV 35S-GFP). Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, No. 107/11.

Author information

Authors and Affiliations

  1. Department of Fruit Tree Sciences, ARO, The Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel

    Alina Sorkina, Gabriel Bardosh, Ifat Fridman, Ludmila Schlizerman, Naftali Zur, Etti Or & Avi Sadka

  2. Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, People’s Republic of China

    Yong-Zhong Liu

  3. The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, 76100, Israel

    Eliezer E. Goldschmidt

  4. Department of Plant Sciences, University of California, One Shields Ave, Davis, CA, 95616, USA

    Eduardo Blumwald

Authors
  1. Alina Sorkina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gabriel Bardosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yong-Zhong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ifat Fridman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ludmila Schlizerman
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Naftali Zur
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Etti Or
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Eliezer E. Goldschmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Eduardo Blumwald
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Avi Sadka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Avi Sadka.

Additional information

Communicated by L. Peña.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PPTX 8190 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sorkina, A., Bardosh, G., Liu, YZ. et al. Isolation of a citrus promoter specific for reproductive organs and its functional analysis in isolated juice sacs and tomato. Plant Cell Rep 30, 1627–1640 (2011). https://doi.org/10.1007/s00299-011-1073-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-011-1073-3

Keywords

Search

Navigation