Plant Molecular Biology Reporter

, Volume 28, Issue 2, pp 193–203

Cloning and Characterization of Prunus serotina AGAMOUS, a Putative Flower Homeotic Gene

  • Xiaomei Liu
  • Joseph M. Anderson
  • Paula M. Pijut
Article

Abstract

Members of the AGAMOUS subfamily of MADS-box transcription factors play an important role in regulating the development of reproductive organs in flowering plants. To help understand the mechanism of floral development in black cherry (Prunus serotina), PsAG (a putative flower homeotic identity gene) was isolated, and its MIKC-type structure was shown to be a homolog of the Arabidopsis thaliana AG gene. It was a single-copy gene in black cherry. A phylogenetic tree derived from the protein sequence indicated PsAG to be a C-function flower homeotic gene with a high similarity to other AG homologs, such as those from Prunus persica and Prunus mume. PsAG met the criteria for AG subfamily gene structure with a typical MIKC structure. In situ hybridization showed that PsAG was expressed mainly in the floral meristem, such as stamen and carpel primordia during the early stage of floral development, and transcript of PsAG accumulated in the tissues of the ovary, stigma, style, and stamens. When the flowers matured, PsAG had enhanced expression in ovary, style, and stigma, with decreased expression in the stamen. PsAG continued to be expressed in the ovule at the late stage of flower development. The developmental patterns of expression were consistent with those of AG and homologs from other species. Both phylogenetic analysis and expression-pattern data suggest that PsAG was the black cherry homolog of Arabidopsis AG. An RNAi construct with a partial PsAG gene was constructed for black cherry transformation.

Keywords

Black cherry Carpel Flowering MADS-box RNAi Stamen 

References

  1. Bielenberg DG, Wang Y, Fan S, Reighard GL, Scorza R, Abbott AG (2004) A deletion affecting several gene candidates is present in the Evergrowing peach mutant. J Hered 95:436–444CrossRefPubMedGoogle Scholar
  2. Boss PK, Vivier M, Matsumoto S, Dry IB, Thomas MR (2001) A cDNA from grapevine (Vitis vinifera L.) shows homology to AGAMOUS and SHATTERPROOF is not only expressed in flowers but also throughout berry development. Plant Mol Biol 45:541–553CrossRefPubMedGoogle Scholar
  3. Bowman JL, Drews GN, Meyerowitz EM (1991a) Expression of the Arabidopsis floral homeotic gene AGAMOUS is restricted to specific cell types late in flower development. Plant Cell 3:749–758CrossRefPubMedGoogle Scholar
  4. Bowman JL, Smyth DR, Meyerowitz EM (1991b) Genetic interactions among floral homeotic genes of Arabidopsis. Development 112:1–20PubMedGoogle Scholar
  5. Brunner AM, Rottmann WH, Sheppard LA, Krutovskii K, DiFazio SP, Leonardi S, Strauss SH (2000) Structure and expression of duplicate AGAMOUS orthologues in poplar. Plant Mol Biol 44:619–634CrossRefPubMedGoogle Scholar
  6. Coen ES, Meyerowitz EM (1991) The war of the whorls: genetic interactions controlling flower development. Nature 353:31–37CrossRefPubMedGoogle Scholar
  7. Davies B, Motte P, Keck E, Saedler H, Sommer H, Schwarz-Sommer Z (1999) PLENA and FARINELLI: redundancy and regulatory interactions between two Antirrhinum MADS-box factors controlling flower development. EMBO J 18:4023–4034CrossRefPubMedGoogle Scholar
  8. Deyholos MK, Sieburth LE (2000) Separable whorl-specific expression and negative regulation by enhancer elements within the AGAMOUS second intron. Plant Cell 12:1799–1810CrossRefPubMedGoogle Scholar
  9. Dower WJ, Miller JF, Ragsdale CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucl Acids Res 16:6127CrossRefPubMedGoogle Scholar
  10. Fan HY, Hu Y, Tudor M, Ma H (1997) Specific interactions between the K domains of AG and AGLs, members of the MADS domain family of DNA binding proteins. Plant J 12:999–1010CrossRefPubMedGoogle Scholar
  11. Hasebe M, We CK, Kat M, Banks JA (1998) Characterization of MADS homeotic genes in the fern Ceratopteris richardii. Proc Natl Acad Sci 95:6222–6227CrossRefPubMedGoogle Scholar
  12. Hong RL, Hamaguchi L, Busch MA, Weigel D (2003) Regulatory elements of the floral homeotic gene AGAMOUS identified by phylogenetic footprinting and shadowing. Plant Cell 15:1296–1309CrossRefPubMedGoogle Scholar
  13. Honma T, Goto K (2001) Complexes of MADS-box proteins are sufficient to convert leaves into floral organs. Nature 409:525–529CrossRefPubMedGoogle Scholar
  14. Jackson D (1991) In-situ hybridization in plants. In: Gurr SJ, McPherson MJ, Bowles DJ (eds) Molecular plant pathology: a practical approach. Oxford University Press, Oxford, pp 163–174Google Scholar
  15. Jager M, Hassanin A, Manuel M, Le Guyader H, Deutsch J (2003) MADS-box genes in Ginkgo biloba and the evolution of the AGAMOUS family. Mol Biol Evol 20:842–854CrossRefPubMedGoogle Scholar
  16. Kater MM, Colombo L, Franken J, Busscher M, Masiero S, Van Lookeren Campagne MM, Angenent GC (1998) Multiple AGAMOUS homologs from cucumber and petunia differ in their ability to induce reproductive organ fate. Plant Cell 10:171–182CrossRefPubMedGoogle Scholar
  17. Kitahara K, Matsumoto S (2000) Rose MADS-box genes ‘MASAKO C1 and D1’ homologous to class C floral identity genes. Plant Sci 151:121–134CrossRefPubMedGoogle Scholar
  18. Kotoda N, Wada M, Komori S, Kidou S, Abe K, Masuda T, Soejima J (2000) Expression pattern of homologues of floral meristem identity genes LFY and AP1 during flower development in apple. J Amer Soc Hort Sci 125:398–403Google Scholar
  19. Kotoda N, Wada M, Kusaba S, Kano-Murakami Y, Masuda T, Soejima J (2002) Overexpression of MdMADS5, an APETALA1-like gene of apple, causes early flowering in transgenic Arabidopsis. Plant Sci 162:679–687CrossRefGoogle Scholar
  20. Kramer EM, Dorit RL, Irish VF (1998) Molecular evolution of gene controlling petal and stamen development: duplicate and divergence within the APETALA3 and PISTILLATA MADS box gene lineages. Genetics 149:765–783PubMedGoogle Scholar
  21. Kramer EM, Jaramillo M, Di Stilio VS (2004) Patterns of gene duplication and functional evolution during the diversification of the AGAMOUS subfamily of MADS box genes in angiosperms. Genetics 166:1011–1023CrossRefPubMedGoogle Scholar
  22. Lemmetyinen J, Hassinen M, Elo A, Porali I, Keinonen K, Makela H, Sopanen T (2004) Functional characterization of SEPALLATA3 and AGAMOUS orthologues in silver birch. Physiol Plant 121:149–162CrossRefPubMedGoogle Scholar
  23. Liu ZR, Liu ZC (2008) The second intron of AGAMOUS drives carpel and stamen-specific expression sufficient to induce complete sterility in Arabidopsis. Plant Cell Rep 27:855–863CrossRefPubMedGoogle Scholar
  24. Liu JY, Huang YH, Ding B, Tauer CG (1999) cDNA cloning and expression of a sweetgum gene that shows homology with Arabidopsis AGAMOUS. Plant Sci 142:73–82CrossRefGoogle Scholar
  25. Martin T, Hu M, Labbé H, McHugh S, Svircev A, Miki B (2006) PpAG1, a homolog of AGAMOUS, expressed in developing peach flowers and fruit. Can J Bot 84:767–776CrossRefGoogle Scholar
  26. Martinez-Castilla LP, Alvarez-Buylla ER (2003) Adaptive evolution in the Arabidopsis MADS-box gene family inferred from its complete resolved phylogeny. Proc Natl Acad Sci USA 100:13407–13412CrossRefPubMedGoogle Scholar
  27. Meilan R, Brunner AM, Skinner JS, Strauss SH (2001) Modification of flowering in transgenic trees. In: Morohoshi N, Komamine A (eds) Molecular Breeding of Woody Plants, pp. 247–256Google Scholar
  28. Parenicová L, de Floter S, Kieffer M, Horner DS, Favalli C, Busscher J, Cook HE, Ingram RM, Kater MM, Davies B, Angenent GC, Colombo L (2003) Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis: new openings to the MADS world. Plant Cell 15:1538–1551CrossRefPubMedGoogle Scholar
  29. Purugganan MD, Rounsley SD, Schmidt RJ, Yanofsky MF (1995) Molecular evolution of flower development: diversification of the plant MADS-box regulatory gene family. Genetics 140:345–356PubMedGoogle Scholar
  30. Rigola D, Pè ME, Fabrizio C, Me G, Sari-Gorla M (1998) CaMADS1, a MADS box gene expressed in the carpel of hazelnut. Plant Mol Biol 38:1147–1160CrossRefPubMedGoogle Scholar
  31. Rutledge R, Regan S, Nicolas O, Fobert P, Cote C, Bosnich W, Kauffeldt C, Sunohara G, Seguin A, Stewart D (1998) Characterization of an AGAMOUS homologue from the conifer black spruce (Picea mariana) that produces floral homeotic conversions when expressed in Arabidopsis. Plant J 15:625–634CrossRefPubMedGoogle Scholar
  32. Salzman RA, Fujita T, Zhu-Salzman K, Hasegawa PM, Bressan RA (1999) An improved RNA isolation method for plant tissues containing high levels of phenolic compounds or carbohydrates. Plant Mol Biol Rep 17:11–17CrossRefGoogle Scholar
  33. Shore P, Sharrocks AD (1995) The MADS-box family of transcription factors. Eur J Biochem 229:1–13CrossRefPubMedGoogle Scholar
  34. Shu GP, Baum DA, Mets LJ (1999) Detection of gene expression patterns in various plant tissues using non-radioactive mRNA in situ hybridization. The World Wide Web J Biology (http://epress.com/w3jbio/vol4/shu/paper.htm)
  35. Sieburth LE, Meyerowitz EM (1997) Molecular dissection of the AGAMOUS control region shows that cis elements for spatial regulation are located intragenically. Plant Cell 9(3):355–365CrossRefPubMedGoogle Scholar
  36. Sieburth LE, Running MP, Meyerowitz EM (1995) Genetic separation of third and fourth whorl functions of AGAMOUS. Plant Cell 7:1249–1258CrossRefPubMedGoogle Scholar
  37. Stairs G, Hauck W (1968) Reproductive cytology of black cherry (Prunus serotina Ehrh.) Proc 15th NE For Tree Improvement Conf, Morgantown, WV, pp. 42-53Google Scholar
  38. Strauss SH, Rottmann WH, Brunner AM, Sheppard LA (1995) Genetic engineering of reproductive sterility in forest trees. Molec Breed 1:5–26CrossRefGoogle Scholar
  39. Sung SK, Yu GH, An G (1999) Characterization of MdMADS2, a member of the SQUAMOSA subfamily of genes, in apple. Plant Physiol 120:969–978CrossRefPubMedGoogle Scholar
  40. Sung SK, Yu GH, Nam J, Jeong DH, An G (2000) Developmentally regulated expression of two MADS-box genes, MdMADS3 and MdMADS4, in the morphogenesis of flower buds and fruits in apple. Planta 210:519–528CrossRefPubMedGoogle Scholar
  41. Swofford DL (2003) PAUP*: phylogenetic analysis using parsimony (* and other methods). Version 4.0b10. Sinauer Associates, SunderlandGoogle Scholar
  42. Tandre K, Svenson M, Svensson ME, Engstrom P (1998) Conservation of gene structure and activity in the regulation of reproductive organ development of conifers and angiosperms. Plant J 15:615–623CrossRefPubMedGoogle Scholar
  43. Theissen G, Saedler H (2001) Plant biology: floral quartets. Nature 409:469–471CrossRefPubMedGoogle Scholar
  44. Vandenbussche M, Theissen G, Van de Peer Y, Gerats T (2003) Structural diversification and neo-functionalization during floral MADS-box gene evolution by C-terminal frameshift mutation. Nucl Acids Res 31:4401–4409CrossRefPubMedGoogle Scholar
  45. van der Linden CG, Vosman B, Smulders MJM (2002) Cloning and characterization of four apple MADS box genes isolated from vegetative tissues. J Exp Bot 53:1025–1036CrossRefPubMedGoogle Scholar
  46. Wada M, Cao Q, Kotoda N, Soejima J, Masuda T (2002) Apple has two orthologues of FLORICAULA/LEAFY involved in flowering. Plant Mol Biol 49:567–577CrossRefPubMedGoogle Scholar
  47. Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, Yin QC, Liu JJ (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27:251–259CrossRefPubMedGoogle Scholar
  48. Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, Green AG, Waterhouse PM (2001) Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J 27:581–590CrossRefPubMedGoogle Scholar
  49. Yanofsky MF, Ma H, Bowman JL, Drews GN, Feldmann KA, Meyerowitz EM (1990) The protein encoded by the Arabidopsis homeotic gene AGAMOUS resembles transcription factors. Nature 346:35–39CrossRefPubMedGoogle Scholar
  50. Yao JL, Dong YH, Kvarnheden A, Morris B (1999) Seven MADS-box genes in apple are expressed in different parts of the fruit. J Amer Soc Hort Sci 124:8–13Google Scholar
  51. Zhang P, Tan HTW, Pwee KH, Kumar PP (2004) Conservation of class C function of floral organ development during 300 million years of evolution from gymnosperms to angiosperms. Plant J 37:566–577CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Xiaomei Liu
    • 1
  • Joseph M. Anderson
    • 2
  • Paula M. Pijut
    • 3
  1. 1.Dept. of Forestry and Natural Resources, Hardwood Tree Improvement and Regeneration Center (HTIRC)Purdue UniversityWest LafayetteUSA
  2. 2.Department of AgronomyUSDA Agricultural Research Service, Purdue UniversityWest LafayetteUSA
  3. 3.Northern Research Station, HTIRCUSDA Forest ServiceWest LafayetteUSA

Personalised recommendations