Embryogenic Pollen Culture: A Promising Target for Genetic Transformation

  • J. Kumlehn

As the immediate product of meiosis, the microspore represents the first stage of pollen development. The haploid chromosome set in addition to embryogenic competence renders microspores one of the most attractive cellular targets for the transfer and stable integration of recombinant DNA into plant genomes. However, the development of methods of plant genetic transformation through gene transfer to embryogenic pollen has turned out to be extremely difficult. In this chapter, diverse gene transfer approaches based upon embryogenic pollen cultures are critically reviewed with regard to target cell type, cell cycle stage, method of gene transfer as well as experimental evidence of transgene integration and homozygosity. Typical constraints and potential pitfalls are discussed and possible strategies to resolve these problems are suggested. Recent demonstrations of the successful generation of instantly true-breeding transgenic plants via gene transfer to embryogenic pollen cultures in both dicotyledonous and monocotyledonous species may encourage further development and broad applications of such methods in research and biotechnology.

Keywords Chimera genetic engineering homozygous gene transfer microspore, transgenic 

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References

  1. Aionesei T, Hosp J,Voronin V, Heberle-Bors E, Touraev A (2006) Methotrexate is a new selectable marker for tobacco immature pollen transformation. Plant Cell Rep 25: 410–416PubMedCrossRefGoogle Scholar
  2. Barakat A, Gallois P, Raynal M, Mestre-Ortega D, Salland C, Guiderdoni E, Delseny M, Bernardi G (2000) The distribution of T-DNA in the genomes of transgenic Arabidopsis and rice. FEBS Lett 471: 161–164PubMedCrossRefGoogle Scholar
  3. Bolik M, Koop HU (1991) Identification of embryogenic microspores of barley (Hordeum vulgare L.) by individual selection and culture and their potential for transformation by microinjection. Protoplasma 162: 61–68CrossRefGoogle Scholar
  4. Carlson AR, Letarte J, Chen J, Kasha KJ (2001) Visual screening of microspore-derived transgenic barley (Hordeum vulgare L.) with green-fluorescent protein. Plant Cell Rep 20: 331–337CrossRefGoogle Scholar
  5. Cheng M, Fry JE, Pang SZ, Zhou HP, Hironaka CM, Duncan DR, Conner TW, Wan YC (1997) Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol 115: 971–980PubMedGoogle Scholar
  6. Dormann M, Wang HM, Oelck M (2001) Transformed embryogenic microspores for the generation of fertile homozygous plants. US Patent 6,316,694 B1Google Scholar
  7. Fennell A, Hauptmann R (1992) Electroporation and PEG delivery of DNA into maize microspores. Plant Cell Rep 11: 567–570CrossRefGoogle Scholar
  8. Ferrie AMR, Keller WA (2007) Optimization of methods for using polyethylene glycol as a non-permeating osmoticum for the induction of microspore embryogenesis in the Brassicaceae. In Vitro Cell Dev Biol — Plant 43: 348–355CrossRefGoogle Scholar
  9. Folling L, Olesen A (2001) Transformation of wheat (Triticum aestivum L.) microspore-derived callus and microspores by particle bombardment. Plant Cell Rep 20: 629–636CrossRefGoogle Scholar
  10. Fukuoka H, Ogawa T, Matsuoka M, Ohkawa Y, Yano H (1998) Direct gene delivery into isolated microspores of rapeseed (Brassica napus L.) and the production of fertile transgenic plants. Plant Cell Rep 17: 323–328CrossRefGoogle Scholar
  11. Gaillard A, Matthys-Rochon E, Dumas C (1992) Selection of microspore derived embryogenic structures in maize related to transformation potential by microinjection. Bot Acta 105: 313–318Google Scholar
  12. González-Melendi P, Ramírez C, Testillano PS, Kumlehn J, Risueno MC (2005) Three dimensional confocal and electron microscopy imaging define the dynamics and mechanisms of diploidisation at early stages of barley microspore-derived embryogenesis. Planta 222: 47–57PubMedCrossRefGoogle Scholar
  13. Hu T, Kasha KJ (1999) A cytological study of pretreatments used to improve isolated microspore cultures of wheat (Triticum aestivum L.) cv. Chris. Genome 3: 432–441CrossRefGoogle Scholar
  14. Huang B (1992) Genetic manipulation of microspores and microspore-derived embryos. In Vitro Cell Dev Biol — Plant 28P: 53–58CrossRefGoogle Scholar
  15. Jaehne A, Becker D, Brettschneider R, Loerz H (1994) Regeneration of transgenic, microspore-derived, fertile barley. Theor Appl Genet 89: 525–533Google Scholar
  16. Jardinaud M-F, Souvré A, Alibert G (1993) Transient GUS gene expression in Brassica napus electroporated microspores. Plant Sci 93: 177–184CrossRefGoogle Scholar
  17. JonesVilleneuve E, Huang B, Prudhomme I, Bird S, Kemble R, Hattori J, Miki B (1995) Assessment of microinjection for introducing DNA into uninuclear microspores of rapeseed. Plant Cell Tiss Org Cult 40: 97–100CrossRefGoogle Scholar
  18. Kuhlmann U, Foroughi-Wehr B, Graner A, Wenzel G (1991) Improved culture system for microspores of barley to become a target for DNA uptake. Plant Breed 107: 165–168CrossRefGoogle Scholar
  19. Kumlehn J, Broeders S, Valkov V (2004) Exclusive generation of true-breeding transgenic plants via Agrobacterium-mediated transformation of barley pollen cultures. Czech J Genet Plant Breed 40: 83Google Scholar
  20. Kumlehn J, Serazetdinova L, Hensel G, Becker D, Loerz H (2006) Genetic transformation of barley (Hordeum vulgare L.) via infection of androgenetic pollen cultures with Agrobacterium tumefaciens. Plant Biotechnol J 4: 251–261PubMedCrossRefGoogle Scholar
  21. Langridge P, Brettschneider R, Lazzeri P, Loerz H (1992) Transformation of cereals via Agrobacterium and the pollen pathway: a critical assessment. Plant J 2: 631–638CrossRefGoogle Scholar
  22. Maluszynski M, Kasha KJ, Szarejko I (2003) Published protocols for other crop plant species. In: Maluszynski M (ed) Doubled Haploid Production in Crop Plants: A Manual. Kluwer Academic Publishers, Dordrecht, Boston, London pp. 309–336Google Scholar
  23. Miki BL, Huang B, Bird S, Kemble R, Simmonds D, Keller W (1989) A procedure for the microinjection of plant cells and protoplasts. J Tiss Cult Methods 12: 139–144CrossRefGoogle Scholar
  24. Nagata Z, Okada K, Takebe I (1986) Strong dependency of the transformation of plant protoplast on cell cycle. In: Kado C (ed) Fallen Leaf Lake conference on Agrobacterium and crown gall. University of California, Davis, p. 9Google Scholar
  25. Neuhaus G, Spangenberg G, Mittelsten G, Schmid O, Schweiger HG (1987) Transgenic rape seed plants obtained by the microinjection of DNA into microspore-derived embryoids. Theor Appl Genet 75: 30–36CrossRefGoogle Scholar
  26. Nishihara M, Seki M, Kyo M, Irifune K, Morikawa H (1995) Transgenic haploid plants of Nicotiana rustica produced by bombardment-mediated transformation of pollen. Transgenic Res 4: 341–348CrossRefGoogle Scholar
  27. Olsen FL (1991) Isolation and cultivation of embryogenic microspores from barley (Hordeum vulgare L.). Hereditas 115: 255–266PubMedCrossRefGoogle Scholar
  28. Pechan PM (1989) Successful cocultivation of Brassica napus microspores and proembryos with Agrobacterium. Plant Cell Rep 8: 387–398CrossRefGoogle Scholar
  29. Potrykus I (1990) Gene transfer to cereals: an assessment. Biotechnology 8: 535–542CrossRefGoogle Scholar
  30. Radchuk V, Borisjuk L, Radchuk R, Steinbiss HH, Rolletschek H, Broeders S, Wobus U (2006) Jekyll encodes a novel protein involved in the sexual reproduction of barley. Plant Cell 18: 1652–1666PubMedCrossRefGoogle Scholar
  31. SalmenkallioMarttila M, Aspegren K, Akerman S, Kurten U, Mannonen L, Ritala A, Teeri TH, Kauppinen J (1995) Transgenic barley (Hordeum vulgare L.) by electroporation of protoplasts. Plant Cell Rep 15: 301–304CrossRefGoogle Scholar
  32. Sangwan RS, Ducrocq C, Sangwan-Norreel B (1993) Agrobacterium-mediated transformation of pollen embryos in Datura innoxia and Nicotiana tabacum — production of transgenic haploid and fertile homozygous dihaploid plants. Plant Sci 95: 99–115CrossRefGoogle Scholar
  33. Shanjun T, Sangwan RS, Raghavan V, Verma DPS, Sangwan-Norreel BS (2005) Transformation of pollen embryo-derived explants by Agrobacterium tumefaciens in Hyoscyamus niger. Plant Cell Tiss Org Cult 81: 139–148CrossRefGoogle Scholar
  34. Shim YS, Kasha KJ (2003) The influence of pretreatment on cell stage progression and the time of DNA synthesis in barley (L.) uninucleate microspores. Plant Cell Rep 21: 1065–1071PubMedCrossRefGoogle Scholar
  35. Stein N, Perovic D, Kumlehn J, Pellio B, Stracke S, Streng S, Ordon F, Graner A (2005) The eukaryotic translation initiation factor 4E confers multiallelic recessive Bymovirus resistance in Hordeum vulgare (L.). Plant J 42: 912–922PubMedCrossRefGoogle Scholar
  36. Stoeger E, Fink C, Pfosser M, Heberle-Bors E (1995) Plant transformation by particle bombardment of embryogenic pollen. Plant Cell Rep 14: 273–278Google Scholar
  37. Swanson EB, Erickson LR (1989) Haploid transformation in Brassica napus using an octopine-producing strain of Agrobacterium tumefaciens. Theor Appl Genet 78: 831–835Google Scholar
  38. Tinland B (1996) The integration of T-DNA into plant genomes. Trends Plant Sci 1: 178–184CrossRefGoogle Scholar
  39. Touraev A, Pfosser M, Heberle-Bors E (2001) The microspore: a haploid multipurpose cell. Adv Bot Res 35: 53–109CrossRefGoogle Scholar
  40. Travella S, Ross SM, Harden J, Everett C, Snape JW, Harwood WA (2005) A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques. Plant Cell Rep 23: 780–789PubMedCrossRefGoogle Scholar
  41. Tzfira T, Citovsky V (2006) Agrobacterium-mediated genetic transformation of plants: biology and biotechnology. Curr Opin Biotechnol 17: 147–154PubMedGoogle Scholar
  42. Villemont E, Dubois F, Sangwan RS, Vasseur G, Bourgeois Y, Sangwan-Norreel BS (1997) Role of the host cell cycle in the Agrobacterium-mediated genetic transformation of Petunia: evidence of an S-phase control mechanism for T-DNA transfer. Planta 201: 160–172CrossRefGoogle Scholar
  43. Vischi M, Marchetti S (1997) Strong extracellular nuclease activity displayed by barley (Hordeum vulgare L.) uninucleate microspores. Theor Appl Genet 95: 185–190CrossRefGoogle Scholar
  44. Wan Y, Lemaux PG (1994) Generation of large numbers of independently transformed fertile barley plants. Plant Physiol 104: 37–48PubMedGoogle Scholar
  45. Yao QA, Simion E, William M, Krochko J, Kasha KJ (1997) Biolistic transformation of haploid isolated microspores of barley (Hordeum vulgare L.). Genome 40: 570–581.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • J. Kumlehn
    • 1
  1. 1.Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)GaterslebenGermany

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