Skip to main content
Log in

Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.)

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. 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).

    Google Scholar 

  2. 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).

    Google Scholar 

  3. Chishlom D: A convenient moderate-scale procedure for obtaining DNA from bacterophage lambda. Biotechniques 7: 21–23 (1989).

    Google Scholar 

  4. 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).

    Google Scholar 

  5. 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).

    Google Scholar 

  6. 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).

    Google Scholar 

  7. 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).

    Google Scholar 

  8. Esau K: Anatomy of Seed Plants, 2nd ed. John Wiley, New York (1977).

    Google Scholar 

  9. 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).

    Google Scholar 

  10. 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).

    Google Scholar 

  11. Goldberg RB, Beals TP, Sanders PM: Anther development: basic principles and practical applications. Plant Cell 5: 1217–1229 (1993).

    Google Scholar 

  12. 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).

    Google Scholar 

  13. 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).

    Google Scholar 

  14. 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).

    Google Scholar 

  15. 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).

    Google Scholar 

  16. 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).

    Google Scholar 

  17. Joshi CP, Zhou H, Huang X, Chiang VL: Context sequences of translation initiation codon in plants. Plant Mol Biol 35: 993–1001 (1997).

    Google Scholar 

  18. Kamalay JC, Goldberg RB: Regulation of structural gene expression in tobacco. Cell 19: 934–46 (1980).

    Google Scholar 

  19. 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).

    Google Scholar 

  20. 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).

    Google Scholar 

  21. 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).

    Google Scholar 

  22. 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).

    Google Scholar 

  23. 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).

    Google Scholar 

  24. Mascarenhas JP: Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338 (1990).

    Google Scholar 

  25. 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).

    Google Scholar 

  26. McCormick S: Molecular analysis of male gametogenesis in plants. Trends Genet 7: 298–303 (1991).

    Google Scholar 

  27. McCormick S: Male gametophyte development. Plant Cell 5: 1265–1275 (1993).

    Google Scholar 

  28. 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).

    Google Scholar 

  29. 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).

    Google Scholar 

  30. 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).

    Google Scholar 

  31. 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).

    Google Scholar 

  32. Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  33. Sanger F, Nicklen S, Coulson AR: DNA sequence with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  34. 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).

    Google Scholar 

  35. Seurinck J, Truettner J, Goldberg RB: The nucleotide sequence of an anther-specific gene. Nucl Acids Res 18: 3403 (1990).

    Google Scholar 

  36. Shen JB, Hsu FC: Brassica anther-specific genes: characterization and in situ localization of expression. Mol Gen Genet 234: 379–389 (1992).

    Google Scholar 

  37. 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).

    Google Scholar 

  38. 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).

    Google Scholar 

  39. 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).

    Google Scholar 

  40. 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).

    Google Scholar 

  41. 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).

    Google Scholar 

  42. Ursin VM, Yamaguchi J, McCormick S: Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1: 727–736 (1989).

    Google Scholar 

  43. 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).

    Google Scholar 

  44. Willing RP, Mascarenhas JP: Analysis of the complexity and diversity of mRNAs from pollen and shoots of Tradescantia. Plant Physiol 75: 865–868 (1984).

    Google Scholar 

  45. 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).

    Google Scholar 

  46. 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).

    Google Scholar 

  47. 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).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006157603096

Navigation