Abstract
The anther-specific cDNA clone Bcp1 from Brassica campestris is expressed in both the haploid pollen and diploid tapetum, as shown by in situ hybridization. We have isolated Bgpl, a genomic clone homologous to Bcpl. The coding region and extensive 5′ flanking sequences of Bgp1 have been sequenced, and the coding region shows 88% identity with Bcp1. RNA gel blot analysis confirmed the expression of Bgp1-specific transcripts in B. campestris pollen. A 767 by 5′ DNA fragment was fused to the reporter gene β-glucuronidase (gus) and introduced into both Arabidopsis thaliana and Nicotiana tabacum by transformation. This 5′ fragment directed high-level expression in the pollen and tapetum of transgenic Arabidopsis. In transgenic tobacco however, the same construct was expressed only in pollen. A series of 5′ deletion constructs has been created and used to transform A. thaliana to analyse the 5′ region of Bgp1. The results indicate that Bgp1 expression in the tapetum and pollen of Arabidopsis requires the presence of different 5′ DNA sequences.
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Albani D, Robert LS, Donaldson PE, Altosaar I, Arnison PG, Fabijanski SF (1991a) Charactization of a pollen-specific gene family from Brassica napus which is activated during early microspore development. Plant Mol Biol 15:605–622
Albani D, Altosaar I, Arnison PG, Fabijanski SF (1991b) A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus. Sequences in its 5′ flanking region are conserved in other pollen-specific promoters. Plant Mol Biol 16:501–513
Coleman A, Goff LJ (1984) Applications of fluorochromes to pollen biology. I. Mithramycin and 4, 6-diamidino-2-phenylindole (DAPI) as a vital stain and quantitation of nuclear DNA. Stain Technol 60:145–154
Hamilton DA, Bashe DM, Stinson JR, Mascarenhas JP (1989) Characterization of a pollen-specific genomic clone from maize. Sex Plant Reprod 2:208–212
Hanson DD, Hamilton DA, Travis JL, Bashe DM, Mascarenhas JP (1989) Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1:173–179
Horsch RB, Fry JE, Hoffmann NL, Wallroth M, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes to plants. Science 227:1229–1231
Jefferson RA, Burgess SM, Hirsh D (1986) β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 83:8447–8451
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
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
Koncz C, Schell J (1986) The promoter of Ti-DNA gene 5 controls the tissue-specific expression of chimeric genes carried by a novel type of Agrobacterium binary vector. Mol Gen Genet 204:383–396
Kozak M (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 4:283–292
Lutcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA (1987) Selection of AUG initiation codons differs in plants and animals. EMBO J 6:43–48
Mascarenhas JP (1988) Anther and pollen expressed genes. In: Verma DPS, Goldberg RB (eds) Temporal and spatial regulation of plants genes. Springer-Verlag, New York, pp 97–125
McCormick S (1991) Molecular analysis of male gametogenesis in plants. Trends Genet 7:298–303
Murray HG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–11325
Sambrook J, Fritsch EF, Maniatis T (1982) Molecular cloning: A laboratory manual. Cold Spring Harbour Laboratory Press, Cold Spring Harbor New York
Sato T, Thorsness MK, Kandasamy MK, Nishio T, Hirai M, Nasrallah J, Nasrallah M (1991) Activity of a S locus gene promoter in pistils and anthers of transgenic Brassica. Plant Cell 3:867–876
Stinson JR, Eisenberg AJ, Willing RP, Enrico Pe M, Hanson DD, Mascarenhas JP (1987) Genes expressed in the male gametophyte of flowering plants and their isolation. Plant Physiol 83:442–447
Theerakulpisut P, Xu HL, Singh MB, Pettitt JM, Knox RB (1991) Isolation and developmental expression of Bepl, an antherspecific cDNA clone in Brassica campestris. Plant Cell 3:1073–1084
Twell D, Wing RA, Yamaguchi J, McCormick S (1989) Isolation and expression of an anther-specific gene from tomato. Mol Gen Genet 217:240–245
Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S (1991) Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev 5:496–507
Valvekens D, Van Montagu M, Van Lijsebettens M (1988) Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc. Natl. Acad. Sci. 85:5536–5540
van der Meer IM, Spelt CE, Mol JNM, Stuitje AR (1990) Promoter analysis of the chalcone synthase (chsA) gene of Petunia hybrida: a 67 by promoter region directs flower-specific expression. Plant Mol. Biol. 15:95–109
Willing RP, Mascarenhas JP (1984) Analysis of the complexity and diversity of mRNAs from pollen and shoots of Tradescantia. Plant Physiol 75:865–868
Willing RP, Bashe D, Mascarenhas JP (1988) An analysis of the quantity and diversity of messenger RNAs from pollen and shoots of Zea mays. Theor Appl Genet 75:751–753
Woodcock DM, Crowther M, Diver WD, Graham M, Bateman C, Baker DJ, Smith SS (1988) Rg1B facilitated cloning of highly methylated eukaryotic DNA: the human L1 transposon, plant DNA and DNA methylated in vitro with human DNA methyl-transferase. Nucleic Acids Res. 16:4465–4482
Xu H (1992) Molecular analysis of anther-specific genes from Brassica campestris and Arabidopsis thaliana. Ph.D. Thesis, University of Melbourne
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Xu, H., Davies, S.P., Kwan, B.Y.H. et al. Haploid and diploid expression of a Brassica campestris anther-specific gene promoter in Arabidopsis and tobacco. Molec. Gen. Genet. 239, 58–65 (1993). https://doi.org/10.1007/BF00281601
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DOI: https://doi.org/10.1007/BF00281601