Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The characterisation of tapetum-specific cDNAs isolated from a Lilium henryi L. meiocyte subtractive cDNA library

  • 49 Accesses

  • 9 Citations


Differential screening of a meiocyte subtractive cDNA library from Lilium henryi L. has identified a group of 16 anther-specific partial cDNAs. Three of these sequences, LHM2, LHM6 and LHM7 have been further characterised. Hybridisation in situ with antisense riboprobes of LHM2, LHM6, and LHM7 gives a strong, clear signal which, contrary to expectations, is localised to the tapetal layer surrounding the meiocytes and not the meiocytes themselves. Developmental slot blots demonstrate that mRNAs corresponding to the three LHM cDNAs are transcribed from prophase of meiosis I to the uninucleate microspore stage, while Northern analysis reveals these tapetally expressed cDNAs to correspond with transcripts of some 500 bp. Although LHM2 is less abundant than LHM6 and LHM7, the pattern of developmental expression, and the size range of the transcripts suggests that all three cDNAs may be related. The deduced polypeptide products of LHM6 and LHM7 share 71% identity over a conserved region of 38 residues. Inverse polymerse chain reaction was used to obtain the full sequence for LHM7. Its deduced protein sequence has a signal peptide indicating it may be secreted; the cleaved protein has a molecular weight of 8.9 kDa. Furthermore, the LHM7 protein has significant levels of homology with tapetally expressed proteins from Arabidopsis thaliana, Antirrhinum majus and Lycopersicon esculentum. All contain a highly conserved pattern of cysteine residues present in seed and non-specific lipid transfer proteins. The function of this gene product is discussed in the perspective of current models of anther development.

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



inverse polymerase chain reaction


non-specific lipid transfer protein


polymerase chain reaction


  1. Albani, D., Altosaar, I., Arnison, P.G., Fabijanski, S.F. (1991) 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

  2. Bennett, M.D., Smith, J.B. (1976) Nuclear DNA amounts in angiosperms. Proc. R. Soc. London Ser. B 274, 227–274

  3. Brown, S.M., Crouch, M.L. (1991) Characterisation of a gene family abundantly expressed in Oenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell 2, 263–274

  4. Chen, R., Smith, A.G. (1993) Nucleotide sequence of a stamen- and tapetum-specific gene from Lycopersicon esculentum. Plant Physiol. 101, 1413

  5. Coen, E.S., Romero, J.M., Doyle, S., Elliott, R., Murphy, G., Carpenter, R. (1990) Floricaula; a homeotic gene required for flower development in Antirrhinum majus. Cell 63, 1311–1322

  6. Dickinson, H.G. (1968) Ultrastructural and physiological aspects of pollen development. Ph.D. thesis, University of Birmingham

  7. Dickinson, H.G. (1981) The structure and chemistry of plastid differentiation during male meiosis in Lilium henryi. J. Cell Sci. 52, 223–241

  8. Dickinson, H.G. (1986) The physiology and biochemistry of meiosis in the anther. Int. Rev. Cytol. 107, 79–109

  9. Dickinson H.G., Bell P.R. (1976) Development of the tapetum in Pinus banksiana preceding sporogenesis. Ann. Bot. 40, 103–113

  10. Dickinson, H.G., Lewis, D. (1973) The formation of the tryphine coating the pollen grains of Raphanus, and its proteins relating to the self-incompatibility system. Proc. R. Soc. London. Ser. B 184, 149–165

  11. Erickson, R.O. (1948) Cytological and growth correlations in the flower bud and anther of Lilium longiflorum. Am. J. Bot. 35, 729–739

  12. Evrard, J-L., Jako, C., Saint-Guily, A., Weil, J-H., Kuntz, M. (1991) Anther-specific, developmentally regulated expression of genes encoding a new class of proline-rich proteins in sunflower. Plant Mol. Biol. 16, 271–281

  13. Greenland, A.J., Thomas, M.V., Walden, R.M. (1987) Expression of two nuclear genes encoding chloroplast proteins during early development of cucumber seedlings. Planta 170, 99–110

  14. Guha, S., Maheshwari, S.C. (1964) In vitro production of embryos from anthers of Datura. Nature 204 497–500

  15. Hanson, D.D., Hamilton, D.A., Travis, J.L., Bashe, D.M., Mascarenhas, J.P. (1989) Characterisation of a pollen-specific genomic clone from maize. Sex. Plant Rep. 2, 208–212

  16. Hass, G.M., Nau, H., Biemann, K., Grahn, D.T., Ericsson, L.H., Neurath, H. (1975) The amino acid sequence of a carboxypeptidase inhibitor from potatoes. Biochemistry 14, 1334–1342

  17. Hass, G.M., Hermodson, M.A. (1981) Amino acid sequence of a carboxypeptidase inhibitor from tomato fruit. Biochemistry 20, 2256–2260

  18. Heslop-Harrison, J. (1968) Tapetal origin of pollen coat substances in Lilium. New Phytol. 67, 779–786

  19. Howley, P.M. (1993) The cloning of microsporogenesis and meiocyte specific sequences from the meiocyte, microspore and pollen cells of Lilium henryi. Ph.D. thesis, University of Reading

  20. Kaul, M.L.H. (1988) Male sterility in higher plants. Monographs on theoretical and applied genetics 10. Springer-Verlag, Berlin

  21. Knox, R.B., Heslop-Harrison, J. (1971) Pollen-wall proteins: electron microscopic localization of acid phosphatase in the intine of Crocus vernus. J. Cell Sci. 8, 727–733

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

  23. Kreis, M., Forde, B.G., Rahman, S., Miflin, B.J., Shewry, P.R. (1985) Molecular evolution of the seed storage proteins of barley, rye and wheat. J. Mol. Biol. 183, 499–502

  24. Maeda, K., Wakabayashi, S., Matsubara, H. (1983) Disulfide bridges in an a-amylase inhibitor from wheat kernel. J. Biochem. 94, 865–870

  25. Mariani, C., De Beuckeleer, M., Truettner, J., Leemans, J., Goldberg, R.B. (1990) Induction of male sterility in plants by a chimeric ribonuclease gene. Nature 347, 737–741

  26. Mascarenhas, J.P. (1990) Gene activity during pollen development. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41, 317–338

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

  28. Murgia, M., Charzynska, M., Cresti, M. (1991) Secretory tapetum of Brassica oleracea L.: polarity and ultrastructural features. Sex. Plant Rep. 4, 28–35

  29. Nacken, W.K.F., Huijser, P., Beltran, J-P., Saedler, H., Sommer, H. (1991a) 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

  30. Nacken, W.K.F., Huijser, P., Saedler, H., Sommer, H. (1991b) Molecular analysis of tap2 an anther-specific gene from Antirrhinum majus. FEBS Lett. 280, 155–158

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

  32. Reznickova, S.A., Dickinson, H.G. (1982) Ultrastructural aspects of storage lipid metabolism in the tapetum of Lilium hybridum var. enchantment. Planta 155, 400–408

  33. Reznickova, S.A., Willemse, T.M. (1980) Formation of pollen in the anther of Lilium II. The function of the surrounding tissues in the formation of pollen and pollen wall. Acta Bot. Neer. 29, 141–156

  34. Sambrook, J., Fritsch, E.F., Maniatis, T. (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press

  35. Scott, R., Dagless, E., Hodge, R., Paul, W., Soufleri, I., Draper, J. (1991) Patterns of gene expression in developing anthers of Brassica napus. Plant Mol. Biol. 17, 195–207

  36. Stern, H., Hotta, Y. (1987) The biochemistry of meiosis. In: Meiosis, pp. 303–331, Moens, P.B., ed. Academic Press, London

  37. Tchang, F.R.G, This, P., Stiefel, V., Arondel, V., Morch, M-D., Pages, M., Puigdomenech, P., Grellet, F., Delseny, M., Bouillon, P., Huet, J-C, Guerbette, F., Beauvais-Cante, F., Duranton, H., Pernollet, J-C., Kader, J-C. (1988) Phospholipid transfer protein: full length cDNA and amino acid sequence in maize. Amino acid sequence homologies between plant phospholipid transfer proteins. J. Biol. Chem. 267, 16849–16855

  38. Terras, F.R.G., Goderis, I.J., Van Leuven, F., Vanderleyden, J., Cammue, B.P.A., Broekaert, W.F. (1992) In vitro antifungal activity of a radish (Raphanus sativus L.) seed protein homologous to nonspecific lipid transfer proteins. Plant Physiol. 100, 1055–1058.

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

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

  41. von Heijne, G. (1983) Patterns of amino acids near signal-sequence cleavage sites. Eur. J. Biochem. 133, 17–21

  42. Wagh, P.V., Bahl, O.P. (1981) Sugar residues on proteins. CRC Crit. Rev. Biochem. 10, 307–377

  43. Willemse, M.T.M., Reznickova, S.A. (1980) Formation of pollen in the anther of Lilium I. Development of the pollen wall. Acta Bot. Neer. 29, 127–140

  44. Wing, R.A., Yamaguchi, J., Larabell, S.K., Ursin, V.M., McCormick, S. (1989) Molecular and genetic characterisation of two pollenexpressed genes that have sequence similarity to pectate lyases of the plant pathogen Erwinia. Plant Mol. Biol. 14, 17–28

  45. Wright, S.Y., Suner, M-M., Bell, P.J., Vaudin, M., Greenland, A.J. (1993) Isolation and characterization of male flower cDNAs from maize. Plant J. 3, 41–49

Download references

Author information

Additional information

This research was supported by a Science and Engineering Research Council Cooperative Awards in Science and Engineering grant in collaboration with ZENECA Seeds, Jealott's Hill. The authors wish to thank Susan Wright, Phil Bell and Ian Jepson from Zeneca Seeds for their help in teaching S.J.C. many of the techniques used.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Crossley, S.J., Greenland, A.J. & Dickinson, H.G. The characterisation of tapetum-specific cDNAs isolated from a Lilium henryi L. meiocyte subtractive cDNA library. Planta 196, 523–529 (1995). https://doi.org/10.1007/BF00203652

Download citation

Key words

  • Anther
  • Anther proteins
  • Cysteine-rich proteins
  • Lilium
  • Tapetum