Abstract
An anther-specific cDNA clone of rice, RA8, was isolated from an anther cDNA library by differential screening. RNA blot analysis indicated that the RA8 transcript is present specifically in anthers and the transcript level increased as flowers matured, reaching the highest level in mature flowers. The RA8 clone contains an open reading frame of 264 amino acid residues with a hydrophobic N-terminal region. The deduced amino acid sequences did not show significant homology to any known sequences. Genomic DNA blot analysis showed that RA8 is a single-copy gene. A genomic clone corresponding to the RA8 cDNA was isolated and its promoter region was fused to the β-glucuronidase (GUS) gene. Transgenic rice plants exhibited anther-specific expression of the GUS reporter gene. Histochemical GUS analysis showed that the RA8 promoter was active in the tapetum, endothecium, and connective tissues of anthers. Experiments showed that expression of the gene starts when microspores are released from tetrads, and it reaches to the maximum level at the late vacuolated-pollen stage. The RA8 promoter may be useful for controlling gene expression in anthers of cereal plants and for generating male-sterile plants.
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Aarts MGM, Hodge R, Kalantidis K, Florack D, Wilson ZA, Mulligan BJ, Stiekema WJ, Scott R, Pereira A: The Arabidopsis male sterility 2 protein shares similarity with reductases in elongation/condensation complexes. Plant J 12: 615–623 (1997).
An G, Ebert PR, Mitra A, Ha SB: Binary vectors. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, pp. A3/1–A3/19. Kluwer Academic Publishers, Dordrecht, Netherlands (1988).
Chishlom D: A convenient moderate-scale procedure for obtaining DNA from bacterophage lambda. Biotechniques 7: 21–23 (1989).
Chung Y-Y, Kim S-R, Finkel D, Yanofsky MF, An G: Early flowering and reduced apical dominance result from ectopic expression of a rice MADS box gene. Plant Mol Biol 26: 657–665 (1994).
Custers JBM, Oldenhof MT, Schrauwen JAM, Cordewener JHG, Wullems GJ, van Lookeren Campagne MM: Analysis of microspore-specific promoters in transgenic tobacco. Plant Mol Biol 35: 689–699 (1997).
Dai Z, Gao J, An K, Lee JM, Edwards GE, An G: Promoter elements controlling developmental and environmental regulation of a tobacco ribosomal protein gene L34. Plant Mol Biol 32: 1055–1065 (1996).
Domon C, Evrard J-L, Herdenberger F, Pillay DTN, Steinmetz A: Nucleotide sequence of two anther-specific cDNAs from sunflower (Helianthus annuus L.). PlantMol Biol 15: 643–646 (1990).
Esau K: Anatomy of Seed Plants, 2nd ed. John Wiley, New York (1977).
Evrard J-L, Jako C, Saint-Guily A, Weil J-H, Kuntz M: Anther-specific, developmentally regulated expression of genes encoding a new class of proline-rich proteins in sun-flower. Plant Mol Biol 16: 271–281 (1991).
Foster GD, Robinson SW, Blumdell RP, Roberts MR, Hodge R, Draper J, Scott RJ: A Brassica napus mRNA encoding a protein homologous to phospholipid transfer proteins, is expressed specifically in the tapetum and developing microspores. Plant Sci 84:187–192 (1992).
Goldberg RB, Beals TP, Sanders PM: Anther development: basic principles and practical applications. Plant Cell 5: 1217–1229 (1993).
Hamilton DA, Roy M, Rueda J, Sindhu RK, Sanford J, Mascarenhas JP: Dissection of a pollen-specific promoter from maize by transient transformation assays. Plant Mol Biol 18: 211–218 (1992).
Hanson DD, Hamilton DA, Travis JL, Bashe DM, Mascarenhas JP: Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1: 173–179 (1989).
Hiei Y, Ohta S, Komari T, Kumashiro T: Efficient transformation of rice (Oryza sativa L) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6: 271–282 (1994).
Hihara Y, Hara C, Uchimiya H: 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 (1996).
Hueros G, Varotto S, Salamini F, Thompson RD: Molecular characterization of BET1, a gene expressed in the endosperm transfer cells of maize. Plant Cell 7: 747–757 (1995).
Joshi CP, Zhou H, Huang X, Chiang VL: Context sequences of translation initiation codon in plants. Plant Mol Biol 35: 993–1001 (1997).
Kamalay JC, Goldberg RB: Regulation of structural gene expression in tobacco. Cell 19: 934–46 (1980).
Kim S-R, Kim Y, An G: Molecular cloning and characterization of anther-preferential cDNA encoding a putative actin-depolymerizing factor. Plant Mol Biol 21: 39–45 (1993).
Kim Y, An G: Pollen-specific expression of the Arabidopsis thaliana α1-tubulin promoter assayed by α-glucuronidase, chloramphenicol acetyltransferase and diphtheria toxin reporter genes. Transgen Res 1: 188–194 (1992).
Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB: Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201–1224 (1990).
Ludwig SR, Oppenheimer DG, Silflow CD, Snudstad DP: the α1-tubulin gene in Arabidopsis thaliana: primary structure and preferential expression in flowers. Plant Mol Biol 10: 311–321 (1988).
Mariani C, de Beuckeleer M, Truettner J, Leemans J, Goldberg RB: Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737–741 (1990).
Mascarenhas JP: Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338 (1990).
Matsunaga S, Kawano S, Takano H, Uchida H, Sakai A, Kuroiwa T: Isolation and developmental expression of male reproductive organ-specific genes in a dioecious campion, Melandrium album (Silene latifolia). Plant J 10: 679–689 (1996).
McCormick S: Molecular analysis of male gametogenesis in plants. Trends Genet 7: 298–303 (1991).
McCormick S: Male gametophyte development. Plant Cell 5: 1265–1275 (1993).
Muschietti J, Dircks L, Vancanneyt G, McCormick S: LAT52 protein is essential for tomato pollen development: pollen expressing antisense LAT52 RNA hydrates and germinates abnormally and cannot achieve fertilization. Plant J 6: 321–338 (1994).
Oldenhof MT, de Groot PFM, Visser JH, Schrauwen JAM, Wullems GJ: Isolation and characterization of a microsporespecific gene from tobacco. Plant Mol Biol 31: 213–225 (1996).
Roberts MR, Bobson F, Foster GD, Draper J, Scott RJ: A Brassica napus mRNA expressed specifically in developing microspores. Plant Mol Biol 17: 295–299 (1991).
Rogers SO, Bendich AJ: Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, pp. A6/1-A6/10. Kluwer Academic Publishers, Dordrecht, Netherlands (1988).
Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequence with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Scott R, Dagless E, Hodge R, Paul W, Soufleri I, Draper J: Patterns of gene expression in developing anthers of Brassica napus. Plant Mol Biol 17: 195–207 (1991).
Seurinck J, Truettner J, Goldberg RB: The nucleotide sequence of an anther-specific gene. Nucl Acids Res 18: 3403 (1990).
Shen JB, Hsu FC: Brassica anther-specific genes: characterization and in situ localization of expression. Mol Gen Genet 234: 379–389 (1992).
Smith AG, Gasser CS, Budelier KA, Fraley RT: Identification and characterization of stamen-and tapetum-specific genes from tomato. Mol Gen Genet 222: 9–16 (1990).
Steinert PM, Mack JW, Korge BP, Gan SO, Haynes SR, Steven AC: Glycine loops in proteins: their occurrence in certain intermediate filament chains, loricrines and single-stranded RNA-binding proteins. Int J Biol Macromol 13: 130–139 (1991).
Theerakulpisut P, Xu H, Singh MB, Pettitt JM, Knox RB: Isolation and developmental expression of Bcp1, an antherspecific cDNA clone in Brassica campestris. Plant Cell 3: 1073–1084 (1991).
Tsuchiya T, Toriyama K, Ejiri S, Hinata K: Molecular characterization of rice genes specifically expressed in the anther tapetum. Plant Mol Biol 26: 1737–1746 (1994).
Twell D, Yamaguchi J, McCormick S: Pollen-specific gene expression in transgenic plants: coordinate regulation of two different tomato gene promoters during microsporogenesis. Development 109: 705–713 (1990).
Ursin VM, Yamaguchi J, McCormick S: Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1: 727–736 (1989).
Willing RP, Bashe D, Mascarenhas JP: An analysis of the quantity and diversity of messenger RNAs from pollen and shoots of Zea mays. Theor Appl Genet 75: 751–753 (1988).
Willing RP, Mascarenhas JP: Analysis of the complexity and diversity of mRNAs from pollen and shoots of Tradescantia. Plant Physiol 75: 865–868 (1984).
Xu H, Davies SP, Kwan BYH, O'Brien AP, Singh M, Knox RB: Haploid and diploid expression of a Brassica campestris anther-specific gene promoter in Arabidopsis and tobacco. Mol Gen Genet 239: 58–65 (1993).
Xu H, Knox RB, Taylor PE, Singh MB: Bcp1, a gene required for male fertility in Arabidopsis. Proc Natl Acad Sci USA 92: 2106–2110 (1995).
Yokoi S, Tsuchiya T, Toriyama K, Hinata K: Tapetum-specific expression of the Osg6B promoter-α-glucuronidase gene in transgenic rice. Plant Cell Rep 16: 363–367 (1997).
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Jeon, JS., Chung, YY., Lee, S. et al. Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.). Plant Mol Biol 39, 35–44 (1999). https://doi.org/10.1023/A:1006157603096
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DOI: https://doi.org/10.1023/A:1006157603096