Skip to main content
SpringerLink
Log in

PCR-generated cDNA library of transition-stage maize embryos: cloning and expression of calmodulin genes during early embryogenesis

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

One hundred maize zygotic embryos microdissected at the transition stage were used to construct a cDNA library after non-selective PCR (NS-PCR) amplification of whole cDNA populations. The library contains 2.3 × 105 recombinants and two different calmodulin cDNAs were cloned using a heterologous probe from petunia. Calmodulin expression was confirmed throughout maize embryogenesis at the mRNA, amplified cDNA and protein levels. Sequence analysis suggests a maize origin for both clones and negligible nucleotide changes linked to PCR. This library is the first described for early plant embryos and represents a breakthrough to isolate genes involved in embryo differentiation.

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

Similar content being viewed by others

References

  1. Abbe EC, Stein OL: The growth of the shoot apex in maize: embryogeny. Am J Bot 41: 285–293 (1954).

    Google Scholar 

  2. Baum G, Chen Y, Arazi T, Takatsuji H, Fromm H: A plant glutamate decarboxylase containing a calmodulin-binding domain: Cloning, sequence and functional analysis. J Bio Chem 268: 19610–19617 (1993).

    Google Scholar 

  3. Braam J, Davis RW: Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell 60: 357–364 (1990).

    Google Scholar 

  4. Breton C, Said C, Dumas C: Cytological and biochemical calendar of ovule and seed development in Populus sp. J Plant Physiol 142: 331–337 (1993).

    Google Scholar 

  5. Clark JK, Sheridan WF: Isolation and characterization of 51 embryo-specific mutations of maize. Plant Cell 3: 935–951 (1991).

    Google Scholar 

  6. de Jong AJ, Schmidt EDL, de Vries SC: Early events in higher-plant embryogenesis: mini review. Plant Mol Biol 22: 367–377 (1993).

    Google Scholar 

  7. Dresselhaus T, Lörz H, Kranz E: Representative cDNA libaries from few plant cells. Plant J 5: 605–610 (1994).

    Google Scholar 

  8. Errampalli D, Patton D, Castle L, Mickelson L, Hansen K, Schnall J, Feldmann K, Meinke D: Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis. Plant Cell 3: 149–157 (1991).

    Google Scholar 

  9. Faure JE, Digonnet C, Dumas C: An in vitro system for adhesion and fusion of maize gametes. Science 263: 1598–1600 (1994).

    Google Scholar 

  10. Fennoy SL, Bailey-Serres J: Synonymous codon usage in Zea mays L. nuclear genes is varied by level of C and G-ending codons. Nucl Acids Res 21: 5294–5300 (1993).

    Google Scholar 

  11. Fischer R, Koller M, Flura M, Mathews S, Page-Strehler M-A, Krebs J, Penniston JT, Carafoli E, Strehler EE: Multiple divergent mRNAs code for a single human calmodulin. J Biol Chem 263: 17055–17062 (1988).

    Google Scholar 

  12. Fromm H, Chua N-H: Cloning of plant cDNAs encoding calmodulin-binding proteins using S35-labeled recombinant calmodulin as a probe. Plant Mol Biol Rep 10: 199–206 (1992).

    Google Scholar 

  13. Froussard P: A random-PCR method (rPCR) to construct whole cDNA library from low amounts of RNA. Nucl Acids Res 20: 2900 (1992).

    Google Scholar 

  14. Gawienowski MC, Szymanski D, Perera IY, Zielinski RE: Calmodulin isoforms in Arabidopsis encoded by multiple divergent mRNAs. Plant Mol Biol 22: 215–225 (1993).

    Google Scholar 

  15. Goldberg RB, Barker SJ, Perez-Grau L: Regulation of gene expression during plant embryogenesis. Cell 56: 149–160 (1989).

    Google Scholar 

  16. Gurr SJ, McPherson MJ: PCR-directed cDNA libraries. In: McPherson MJ, Quirke P, Taylor GR (eds) PCR: A Practical Approach, pp. 147–170. Oxford University Press, Oxford New-York Tokyo (1991).

    Google Scholar 

  17. Heslop-Harrison J, Heslop-Harrison Y: Evaluation of pollen viability by enzymatically induced fluorescence: Intracellular hydrolysis of fluorescein diacetate. Stain Technol 45: 115–120 (1970).

    Google Scholar 

  18. Jena PK, Reddy ASN, Poovaiah BW: Molecular cloning and sequencing of a cDNA for plant calmodulin: Signal-induced changes in the expression of calmodulin. Proc Natl Acad Sci USA 86: 3644–3648 (1989).

    Google Scholar 

  19. Jepson I, Bray J, Jenkins G, Schuch W, Edwards K: A rapid procedure for the construction of PCR cDNA libraries from small amounts of plant tissue. Plant Mol Biol Rep 9: 131–138 (1991).

    Google Scholar 

  20. Jürgens G, Mayer U, Torres Ruiz RA, Berleth T, Misera S: Genetic analysis of pattern formation in the Arabidopsis embryo. Development S1: 27–38 (1991).

    Google Scholar 

  21. Kranz E, Lörz H: In vitro fertilization with isolated single gametes results in zygotic embryogenesis and fertile maize plants. Plant Cell 5: 739–746 (1993).

    Google Scholar 

  22. Leshem YY: The role of calcium in the membrane. In: Leshem YY (ed) Plant Membranes: A Biophysical approach to Structure, Development and Senescence, pp. 157–173. Kluwer Academic Publishers. Dordrecht (1992).

    Google Scholar 

  23. Luehrsen KR, Walbot V: The impact of AUG start codon context on maize gene expression in vivo. Plant Cell Rep 13: 454–458 (1994).

    Google Scholar 

  24. Mol R, Matthys-Rochon E, Dumas C: In-vitro culture of fertilized embryo sacs of maize: zygotes and two-celled proembryos can develop into plants. Planta 189: 213–217 (1993).

    Google Scholar 

  25. Oh SH, Steiner HY, Dougall DK, Roberts DM: Modulation of calmodulin levels, calmodulin methylation, and calmodulin binding proteins during carrot cell growth and embryogenesis. Arch Biochem Biophys 297: 28–34 (1992).

    Google Scholar 

  26. Rogers SO, Bendich AJ: Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant Molecular Biology Manual., pp. A6/1-A6/11. Kluwer Academic Publishers, Dordrecht (1988).

    Google Scholar 

  27. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  28. Smith DE, Gridley T: Differential screening of a PCR-generated mouse embryo cDNA library: glucose transporters are differentially expressed in early postimplantation mouse embryos. Development 116: 555–561 (1992).

    Google Scholar 

  29. Tam AW, Smith MM, Fry KE, Larrick JW: Construction of cDNA libraries from small numbers of cells using independent primers. Nucl Acids Res 16: 1269 (1989).

    Google Scholar 

  30. Timmers ACJ, de Vries SC, Schel JHN: Distribution of membrane-bound calcium and activated calmodulin during somatic embryogenesis of carrot (Daucus carota L.). Protoplasma 153: 24–29 (1989).

    Google Scholar 

  31. Wada K, Wada Y, Ishibashi F, Gojobori T, Ikemura T: Codon usage tabulated from the GenBank genetic sequence data. Nucl Acids Res 20: 2111–2118 (1992).

    Google Scholar 

  32. Watillon B, Kettmann R, Boxus P, Burny A: A calcium/calmodulin-binding serine/threonine protein kinase homologous to the mammalian type II calcium/calmodulin-dependent protein kinase is expressed in plant cells. Plant Physiol 101: 1381–1384 (1993).

    Google Scholar 

Download references

Author information

Author notes

  1. Christian Breton

    Present address: Centre de Recherches Forestières, I.N.R.A. Orléans, 45160, Ardon, France

Authors and Affiliations

  1. Reconnaissance Cellulaire et Amélioration des Plantes-CNRS-INRA, E.N.S. Lyon, 46 allée d'Italie, 69364, Lyon Cedex 07, France

    Christian Breton, Annie Chaboud, Elisabeth Matthys-Rochon, Elizabeth E. M. Bates, J. Mark Cock & Christian Dumas

  2. Department of Plant Genetics, Weizmann Institute of Science, 76100, Rehovot, Israël

    Hillel Fromm

Authors
  1. Christian Breton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Annie Chaboud
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elisabeth Matthys-Rochon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elizabeth E. M. Bates
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Mark Cock
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hillel Fromm
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christian Dumas
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Breton, C., Chaboud, A., Matthys-Rochon, E. et al. PCR-generated cDNA library of transition-stage maize embryos: cloning and expression of calmodulin genes during early embryogenesis. Plant Mol Biol 27, 105–113 (1995). https://doi.org/10.1007/BF00019182

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00019182

Key words

Search

Navigation