Plant Molecular Biology

, Volume 30, Issue 6, pp 1181–1193 | Cite as

Isolation and characterization of two cDNA clones for mRNAs that are abundantly expressed in immature anthers of rice (Oryza sativa L.)

  • Yukako Hihara
  • Chikage Hara
  • Hirofumi Uchimiya
Research Article


The relationship between the length of anthers and the stage of development of microspores was examined in rice (Oryza sativa L. cv. Hayayuki). Anthers of ≤2 mm and 2.1–2.2 mm in length and those ready to dehiscence were determined to be at the uninucleate, binucleate and trinucleate microspore stage, respectively.

Two cDNAs (YY1 and YY2), representing genes that are specifically expressed in anthers at the uninucleate microspore stage, were isolated and characterized. YY1 cDNA encoded an open reading frame of 95 amino acids. Eight cysteine residues with the potential to form disulfide bridges were present in the amino acid sequence. There was a hydrophobic region at the N-terminus of the putative protein, suggesting that the YY1 protein might be secreted. This cysteine motif and the hydrophobic N-terminus are conserved among products of several anther-specific genes or cDNAs isolated from various plant species. These proteins are thought to form a superfamily of proteins that are confined to anthers. The YY1 transcript was localized in the tapetal cells and the peripheral cells of the vascular bundle. YY2 cDNA encoded an open reading frame of 389 amino acids and the deduced amino acid sequence exhibited substantial homology to that of chalcone synthase. Expression of YY2 mRNA was confined to the tapetal cells. The genes correspond to YY1 and YY2 cDNAs were shown to exist as single copies in the rice genome.

Key words

anther development cDNA chalcone synthase tapetum rice 


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  1. 1.
    Brown SM, Crouch ML: Characterization of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell 2: 263–274 (1990).CrossRefPubMedGoogle Scholar
  2. 2.
    Cox KH, Goldberg RB: Analysis of plant gene expression. In: Shaw CH (ed) Plant Molecular Biology: A Practical Approach, pp. 1–34. IRL Press, Oxford (1988).Google Scholar
  3. 3.
    Dixon RA: The phytoalexin response: eliciting, signalling and control of host gene expression. Biol Rev 61: 239–291 (1986).CrossRefGoogle Scholar
  4. 4.
    Drews GN, Bowman JL, Meyerowits EM: Negative regulation of the Arabidopsis homeotic gene AGAMOUS by the APETALA2 product. Cell 65: 991–1002 (1991).CrossRefPubMedGoogle Scholar
  5. 5.
    von Heijne G: A new method for predicting signal sequence cleavage sites. Nucl Acids Res 14: 4683–4690 (1986).PubMedGoogle Scholar
  6. 6.
    Heller W, Hahlbrock K: Highly purified ‘flavanone synthase’ from parsley catalyzes the formation of naringenin chalcone. Arch Biochem Biophys 200: 617–619 (1980).PubMedGoogle Scholar
  7. 7.
    Kehrel B, Wiermann R: Immunochemical localization of phenylalanine ammonia-lyase and chalcone synthase in anthers. Planta 163: 183–190 (1985).Google Scholar
  8. 8.
    Kobayashi T, Kobayashi E, Sato S, Hotta Y, Miyajima N, Tanaka A, Tabata S: Characterization of cDNAs induced in meiotic prophase in lily microsporocytes. DNA Res 1: 15–26 (1994).PubMedGoogle Scholar
  9. 9.
    Koes RE, van Blokland R, Quattrocchio F, van Tunen AJ, Mol JNM: Chalcone synthase promoters in petunia are active in pigmented and unpigmented cell types. Plant Cell 2: 379–392 (1990).CrossRefPubMedGoogle Scholar
  10. 10.
    Koltunow AW, 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).CrossRefPubMedGoogle Scholar
  11. 11.
    Kreis M, Forde BG, Rahman S, Miflin BJ, Shewry PR: Molecular evolution of the seed storage proteins of barley, rye and wheat. J Mol Biol 183: 499–502 (1985).CrossRefPubMedGoogle Scholar
  12. 12.
    Lilley GG, Inglis A: Amino acid sequence of conglutin δ, a sulphur-rich seed protein of Lupinus angustifolius L. FEBS Lett 195: 235–241 (1986).CrossRefGoogle Scholar
  13. 13.
    Maeda K, Wakabayashi S, Matsubara H: Disulphide bridges in an α-amylase inhibitor from wheat kernel. J Biochem 94: 865–870 (1983).PubMedGoogle Scholar
  14. 14.
    Mascarenhas JP: The biochemistry of angiosperm pollen development. Bot Rev 41: 259–311 (1975).Google Scholar
  15. 15.
    Mascarenhas JF: Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338 (1990).CrossRefGoogle Scholar
  16. 16.
    Mascarenhas JP: Pollen gene expression: molecular evidence. Int Rev Cytol 140: 3–18 (1992).PubMedGoogle Scholar
  17. 17.
    McCormick SM: Molecular analysis of male gametogenesis in plants. Trends Genet 7: 298–303 (1991).PubMedGoogle Scholar
  18. 18.
    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).CrossRefPubMedGoogle Scholar
  19. 19.
    Nacken WKF, Huijser P, Beltran JP, Saedler H, Sommer H: 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 (1991).PubMedGoogle Scholar
  20. 20.
    Niesbach-Klösgen U, Barzen E, Bernhardt J, Rohde W: Chalcone synthase genes in plants: A tool to study evolutionary relationships. J Mol Evol 26: 213–225 (1987).Google Scholar
  21. 21.
    Palmiter RD: Magnesium precipitation of ribonucleoprotein complexes: Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry 13: 3606–3615 (1974).PubMedGoogle Scholar
  22. 22.
    Paul W, Hodge R, Smartt S, Draper J, Scott R: The isolation and characterization of the tapetum-specific Arabidopsis thaliana A9 gene. Plant Mol Biol 19: 611–622 (1992).PubMedGoogle Scholar
  23. 23.
    Raghavan V: Anther and pollen development in rice (Oryza sativa). Am J Bot 75: 183–196 (1988).Google Scholar
  24. 24.
    Reimold U, Kröger M, Kreuzaler F, Hahlbrock K: Coding and 3′ non-coding nucleotide sequence of chalcone synthase mRNA and assignment of amino acid sequence of the enzyme. EMBO J 2: 1801–1805 (1983).Google Scholar
  25. 25.
    Satake T: Male sterility caused by cooling treatment at the young microspore stage in rice plants. IX. Revision of the classification and terminology of pollen developmental stages. Proc Crop Sci Soc Japan 43: 31–35 (1974).Google Scholar
  26. 26.
    Satake T, Hayase H: Male sterility caused by cooling treatment at the young microspore stage in rice plants. V. Estimations of pollen developmental stage and the most sensitive stage to coolness. Japan J Crop Sci 50: 495–501 (1981).Google Scholar
  27. 27.
    Schröder G, Brown JWS, Schröder J: Molecular analysis of resveratrol synthase: cDNA genemic clones and relationship with chalcone synthase. Eur J Biochem 172: 161–169 (1988).PubMedGoogle Scholar
  28. 28.
    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).PubMedGoogle Scholar
  29. 29.
    Shen JB, Hsu FC: Brassica anther-specific genes: characterization and in situ localization of expression. Mol Gen Genet 234: 379–389 (1992).PubMedGoogle Scholar
  30. 30.
    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).PubMedGoogle Scholar
  31. 31.
    Staiger D, Kappeler S, Müller M, Apel K: The proteins encoded by two tapetum-specific transcripts, Sa tap 35 and Sa tap 44, from Sinapis alba L. are localized in the exine cell wall layer of developing microspores. Planta 192: 221–231 (1994).CrossRefPubMedGoogle Scholar
  32. 32.
    Tsuchiya T, Toriyama K, Nasrallah ME, Ejiri S: Isolation of genes abundantly expressed in rice anthers at the microspore stage. Plant Mol Biol 20: 1189–1193 (1992).CrossRefPubMedGoogle Scholar
  33. 33.
    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).PubMedGoogle Scholar
  34. 34.
    van der Meer IM, Stam ME, van Tunen AJ, Mol JNM: Antisense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell 4: 253–262 (1992).PubMedGoogle Scholar
  35. 35.
    Wada T, Ito T, Ito M, Takeoka Y: Light microscopic observation on pollen and anther in rice (Oryza sativa L.). II. Stages from early microspore to mature pollen. Jpn J Crop Sci 61: 136–144 (1992).Google Scholar
  36. 36.
    Wiermann R, Vieth K: Outer pollen wall, an important accumulation site for flavonoids. Protoplasma 118: 230–233 (1983).Google Scholar
  37. 37.
    Wing RA, Yamaguchi J, Larabell SK, Ursin VM, McCormick S: Molecular and genetic characterization of two pollen-expressed genes that gave sequence similarity to pectate lyases of the plant pathogen Erwinia. Plant Mol Biol 14: 17–28 (1990).CrossRefPubMedGoogle Scholar
  38. 38.
    Wright SY, Suner MM, Bell PJ, Vaudin M, Greenland AJ: Isolation and characterization of male flower cDNAs from maize. Plant J 3: 41–49 (1993).CrossRefPubMedGoogle Scholar
  39. 39.
    Ylstra B, Busscher J, Franken J, Hollmen PCH, Mol JNM, van Tunen AJ: Flavonols and fertilization in Petunia hybrida: localization and mode of action during pollen tube growth. Plant J 6: 201–212 (1994).CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Yukako Hihara
    • 1
  • Chikage Hara
    • 1
  • Hirofumi Uchimiya
    • 1
  1. 1.Institute of Molecular and Cellular BiosciencesUniversity of TokyoBunkyo-ku, TokyoJapan

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