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Isolation of rapeseed genes expressed early and specifically during development of the male gametophyte

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Abstract

A cDNA subtraction and differential hybridization strategy was used to isolate cDNAs expressed early during male gametophyte development in the important crop species Brassica napus. Three cDNAs, corresponding to genes highly and specifically expressed at the tetrad and microspore stages, are presented here. The analysis of one of them, named BnM3.4, by in situ hybridization, showed that it is expressed specifically and at a high level in the rapeseed microspore. The specificity in its profile of expression is most likely transcriptionally controlled as a similar pattern of expression was also observed in Arabidopsis thaliana plants transformed by the BnM3.4 promoter fused to the reporter GUS-coding sequence. The putative BnM3.4 promoter contains three dispersed copies of a motif described previously in the promoters of several genes expressed in the male gametophyte. The BnM3.4 gene encodes a predicted novel proline-rich protein of 23.4 kDa which may interact with cytoskeletal components or have a structural role in the cell wall.

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References

  • Aguirre, P.J. and Smith, A.G. 1993. Molecular characterization of a gene encoding a cysteine-rich protein preferentially expressed in anthers of Lycopersicon esculentum. Plant Mol. Biol. 23: 477-487.

    Google Scholar 

  • Albani, D., Robert, L.S., Donaldson, P.A., Altosaar, I., Arnison, P.G. and Fabijanski, S.F. 1990. Characterization of a pollen-specific gene family from Brassica napus which is activated during early microspore development. Plant Mol. Biol. 15: 605-622.

    Google Scholar 

  • Albani, D., Sardana, R., Robert, L.S., Altosaar, I., Arnison, P.G. and Fabijanski, S.F. 1992. A Brassica napus gene family which shows sequence similarity to ascorbate oxidase is expressed in developing pollen. Molecular characterization and analysis of promoter activity in transgenic tobacco plants. Plant J. 2: 331-342.

    Google Scholar 

  • Bechtold, N., Ellis, J. and Pelletier, G. 1993. In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C.r. Acad. Sci. (Sér. III: Sci. Vie) 316: 1194-1199.

    Google Scholar 

  • Bedinger, P.A., Hardeman, K.J. and Loukides, C.A. 1994. Travelling in style: the cell biology of pollen. Trends Cell Biol. 4: 132-138.

    Google Scholar 

  • Bevan, M., Barnes, W.M. and Chilton, M.-D. 1983. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucl. Acids Res. 11: 369-385.

    Google Scholar 

  • Bonhomme, S., Horlow, C., Vezon, D., de Laissardière, S., Guyon, A., Férault, M., Marchand, M., Bechtold, N. and Pelletier, G. 1999. T-DNA mediated disruption of essential gametophytic genes in Arabidopsis is unexpectedly rare and cannot be inferred from segregation distortion alone. Mol. Gen. Genet, in press.

  • Bowman, J. (Ed.) 1994. Arabidopsis. An Atlas of Morphology and Development, Springer-Verlag New York, pp. 133-332.

  • Brander, K.A. and Kuhlemeier, C. 1995. A pollen-specific DEADbox protein related to translation initiation factor eIF-4A from tobacco. Plant Mol. Biol. 27: 637-649.

    Google Scholar 

  • Brown, S.M. and Crouch, M.L. 1990. Characterization of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell 2: 263-274.

    Google Scholar 

  • Bucciaglia, P.A. and Smith, A.G. 1994. Cloning and characterization of Tag1, a tobacco anther _-1,3-glucanase expressed during tetrad dissolution. Plant Mol. Biol. 24: 903-914.

    Google Scholar 

  • Cartea, M.E., Migdal, M., Galle, A.M., Pelletier, G. and Guerche, P. 1998 Comparison of sense and antisense methodologies for modifying the fatty acid composition of Arabidopsis thaliana oilseed. Plant Sci 136: 181-194.

    Google Scholar 

  • Chen, C.G., Mau, S.L. and Clarke, A.E. 1993. Nucleotide sequence and style-specific expression of a novel prolin-rich protein gene from Nicotiana alata. Plant Mol. Biol. 21: 391-395.

    Google Scholar 

  • Connelly, C. and Heiter, P. 1996. Budding yeast SKP1 encodes an evolutionarily conserved kinetochore protein required for cell cycle progression. Cell 86: 275-285.

    Google Scholar 

  • Chen, R. and Smith, A.G. 1993. Nucleotide sequence of a stamenand tapetum-specific gene from Lycopersicon esculentum. Plant Physiol. 101: 1413.

    Google Scholar 

  • Dean, C., van den Elzen, P., Tamaki, S., Dunsmuir, P. and Bedsbrook, J. 1985. Differential expression of eight genes of petunia ribulose bisphosphate carboxylase small subunit multi-gene family. EMBO J. 4: 3055-3061.

    Google Scholar 

  • Dellaporta, J.L., Wood, J. and Hicks, J.B. 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep. 1: 19-21.

    Google Scholar 

  • Estelle, M.A. and Sommerville, C.R. 1987. Auxin-resistant mutants of Arabidopsis thaliana with an altered morphology. Mol. Gen. Genet. 206: 200-206.

    Google Scholar 

  • Feinberg, A.P. and Vogelstein, B. 1985. A technique for radiolabelling DNA fragments to high specificity. Anal. Biochem. 137: 266-267.

    Google Scholar 

  • Feldmann, K., Coury, D. and Christianson, M. 1997. Exceptional segregation of a selectable marker (KanR) in Arabidopsis identi-fies genes important for gametophytic growth and development. Genetics 147: 1411-1422.

    Google Scholar 

  • Gourret, J.P., Delourme, R. and Renard, M. 1992. Expression of ogu cytoplasmic male sterility in cybrids of Brassica napus. Theor. Appl. Genet. 83: 549-556.

    Google Scholar 

  • Hanson, D.D., Hamilton, D.A., Travis, J.L., Bashe, D.M. and Mascarenhas, J.P. 1989. Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1: 173-179.

    Google Scholar 

  • Hebsgaard, S.M., Korning, P.G., Tolstrup, N., Engelbrecht, J., Rouze, P. and Brunak, S. 1996. Splice site prediction in Arabidopsis thaliana pre-mRNA by combining local and global sequence information. Nucl. Acids Res. 24: 3439-3452.

    Google Scholar 

  • Hihara, Y., Hara, C. and Uchimiya, H. 1996. Isolation and characterization of two cDNA clones for mRNAs that are abundantly expressed in immature anthers of rice (Oryza sativa L.). Plant Mol. Biol. 30: 1181-1193.

    Google Scholar 

  • Jackson, D.P. 1991. In situ hybridization in plants. In: D.J. Bowles, S.J. Gurr and M. McPhereson (Eds.), Molecular Plant Pathology: A Practical Approach, Oxford University Press, Oxford, pp. 163-174.

    Google Scholar 

  • Jefferson, R.A., Burgess, S.M. and Hirsh, D. 1986. ß-glucuronidase from Escherichia coli as a gene-fusion marker. Proc. Natl. Acad. Sci. USA 83: 8447-8451.

    Google Scholar 

  • Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: ß-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901-3907.

    Google Scholar 

  • Joshi, C.P. 1987a. An inspection of the domain between putative TATAbox and translation start site in 79 plant genes. Nucl. Acids Res. 15: 6643-6653.

    Google Scholar 

  • Joshi, C.P. 1987b. Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl. Acids Res. 15: 9627-9640.

    Google Scholar 

  • Joshi, C.P., Zhou, H., Huang, X. and Chiang, V.L. 1987. Context sequences of translation initiation codon in plants. Plant Mol. Biol. 35: 993-1001.

    Google Scholar 

  • Kobayashi, T., Kobayashi, E., Sato, S., Hotta, Y., Miyajima, N., Tanaka, A. and Tabata, S. 1994. Characterization of cDNAs induced in meiotic prophase in lily microsporocytes. DNA Res. 1: 15-26.

    Google Scholar 

  • Koltunow, A.M., Truettner, J., Cox, K.H., Wallroth, M. and Goldberg, R.B. 1990. Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201-1224.

    Google Scholar 

  • Koncz, C. and Schell, J. 1986. The promoter of TL-DNA gene 50 controls the tissue specific expression of chimeric genes carried by a novel type of Agrobacterium binary vector. Mol. Gen. Genet. 204: 383-396.

    Google Scholar 

  • Krek, W. 1998. Proteolysis and the G1-S transition: the SCF connection. Curr. Opin. Genet. Devel. 8: 36-42.

    Google Scholar 

  • Kröger, N., Bergsdorf, C. and Sumper, M. 1996. Frustulins: domain conservation in a protein family associated with diatom cell walls. Eur. J. Biochem. 239: 259-264.

    Google Scholar 

  • Lo, Y.-M.D., Mehal, W.Z. and Kleming, K.A. 1988. Rapid production of free biotinylated probes using the polymerase chain reaction. Nucl. Acids Res. 16: 8719-8722.

    Google Scholar 

  • Messing, J., Geraghty, D., Heidecker, G., Hu, N.T., Kridl, J. and Rubenstein, I. 1983. Plant gene structure. In: T. Kosuge, C.P. Meredith and A. Hollaender (Eds.), Genetic Engineering of Plants, Plenum, New York, pp. 211-227.

    Google Scholar 

  • Mueller, P.R. and Wold, B. 1989. In vivo footprinting of a musclespecific enhancer by ligation mediated PCR. Science 246: 780-786.

    Google Scholar 

  • Nacken, W.K.F., Huijser, P., Beltran, J.-P., Saedler, H. and Sommer, H. 1991. Molecular characterization of two stamen-specific genes, tap1 and fil1, that are expressed in the wild type, but not in the deficiens mutant of Antirrhinum majus. Mol. Gen. Genet. 229: 129-136.

    Google Scholar 

  • Nagel, R., Masel, E.A., Birch, R.G. and Manners, J.M. 1990. Electroporation of binary Ti plasmid vector into Agrobacterium tumefaciens and Agrobacterium rhizogenes. FEMS Microbiol. Lett. 67: 325-328.

    Google Scholar 

  • Nielsen, H., Engelbrecht, J., Brunak, S., Von Heijne, G. 1997. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Prot. Engng. 10: 1-6.

    Google Scholar 

  • Oldenhof, M.T., de Groot, P.F.M., Visser, J.H., Schrauwen, J.A.M. and Wullems, G.J. 1996. Isolation and characterization of a microspore-specific gene from tobacco. Plant Mol. Biol. 31: 213-225.

    Google Scholar 

  • Paul, W., Hodge, R., Smartt, S., Draper, J. and Scott, R. 1992. The isolation and characterisation of the tapetum-specific Arabidopsis thaliana A9 gene. Plant Mol. Biol. 19: 611-622.

    Google Scholar 

  • Robert, L.S., Allard, S., Gerster, J.L., Cass, L. and Simmonds, J. 1993. Isolation and characterization of a polygalacturonase gene highly expressed in Brassica napus pollen. Plant Mol. Biol. 23: 1273-1278.

    Google Scholar 

  • Ross, J.H.E. and Murphy, D.J. 1996. Characterization of antherexpressed genes encoding a major class of extracellular oleosinlike proteins in the pollen coat of Brassicaceae. Plant J. 9: 625-637.

    Google Scholar 

  • Rubinelli, P., Hu, Y. and Ma, H. 1998. Identification, sequence analysis and expression studies of novel anther-specific genes of Arabidopsis thaliana. Plant Mol. Biol. 37: 607-619.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Scott, R., Dagless, E., Hodge, R., Paul, W., Soufleri, I. and Draper, J. 1991. Patterns of gene expression in developing anthers of Brassica napus. Plant Mol. Biol. 17: 195-207.

    Google Scholar 

  • Shen, J.B. and Hsu, F.C. 1992. Brassica anther-specific genes: characterization and in situ localization of expression. Mol. Gen. Genet. 234: 379-389.

    Google Scholar 

  • Singh, A., Kao, T. and Lin, J.J. 1993. Transformation of Agrobacterium tumefaciens with T-DNA vectors using high-voltage electroporation. Focus 15: 84-87.

    Google Scholar 

  • Sive, H.L. and St John, T. 1988. A simple subtractive hybridization technique employing photoactivatable biotin and phenol extraction. Nucl. Acids Res. 16: 10937.

    Google Scholar 

  • Stanchev, B.S., Doughty, J., Scutt, C.P., Dickinson, H. and Croy R.R.D. 1996. Cloning of PCP1, a member of a family of pollen coat protein (PCP) genes from Brassica oleracea encoding novel cysteine-rich proteins involved in pollen-stigma interactions. Plant J. 10: 303-313.

    Google Scholar 

  • Theerakulpisut, P., Xu, H., Singh, M.B., Pettitt, J.M. and Knox, R.B. 1991. Isolation and developmental expression of Bcp1, an anther-specific cDNA clone in Brassica campestris. Plant Cell 3: 1073-1084.

    Google Scholar 

  • Twell, D. 1994. The diversity and regulation of gene expression in the pathway of male gametophyte development. In: R.J. Scott and A.D. Stead (Eds.), Molecular and Cellular Aspects of Plant Reproduction, Society for Experimental Biology, Cambridge University, Press, Cambridge, UK, pp. 83-135.

    Google Scholar 

  • Ursin, V.M., Yamaguchi, J. and McCormick, S. 1989. Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1: 727-736.

    Google Scholar 

  • Wang, Z. and Brown, D.B. 1991. A gene expression screen. Proc. Natl. Acad. Sci. USA 88: 11505-11509.

    Google Scholar 

  • Weterings, K., Reijnen, W., van Aarssen, R., Kortstee, A., Spijkers, J., van Herpen, M., Schrauwen, J. and Wullems, G. 1992. Characterization of a pollen-specific cDNA clone fromNicotiana 872 tabacum expressed during microgametogenesis and germination. Plant Mol. Biol. 18: 1101-1111.

    Google Scholar 

  • Weterings, K., Schrauwen, J., Wullems, G. and Twell, D. 1995. Functional dissection of the promoter of the pollen-specific gene NTP303 reveals a novel pollen-specific, and conserved cis-regulatory element. Plant J. 8: 55-63.

    Google Scholar 

  • Woessner, J.P., Molendijk, A.J., van Egmond, P., Klis, F.M., Goodenough, U.W. and Haring, M.A. 1994. Domain conservation in several volvocalean cell wall proteins. Plant Mol. Biol. 26: 947-960.

    Google Scholar 

  • Yu, L.-X., Nasrallah, J., Valenta, R. and Parthasarathy, M.V. 1998. Molecular cloning and mRNA localization of tomato pollen profilin. Plant Mol. Biol. 36: 699-707.

    Google Scholar 

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  1. Station de Génétique et d'Amélioration des Plantes, Institut National de la Recherche Agronomique, Route de Saint Cyr, 78026 , Versailles cedex, France

    Agnès Fourgoux-Nicol, Jan Drouaud, Najat Haouazine, Georges Pelletier & Philippe Guerche

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  1. Agnès Fourgoux-Nicol
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  2. Jan Drouaud
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  3. Najat Haouazine
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  4. Georges Pelletier
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  5. Philippe Guerche
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Fourgoux-Nicol, A., Drouaud, J., Haouazine, N. et al. Isolation of rapeseed genes expressed early and specifically during development of the male gametophyte. Plant Mol Biol 40, 857–872 (1999). https://doi.org/10.1023/A:1006282507095

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