Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 104, Issue 3, pp 283–300 | Cite as

Anther culture for haploid and doubled haploid production



Haploids are plants with a gametophytic chromosome number and doubled haploids are haploids that have undergone chromosome duplication. The production of haploids and doubled haploids (DHs) through gametic embryogenesis allows a single-step development of complete homozygous lines from heterozygous parents, shortening the time required to produce homozygous plants in comparison with the conventional breeding methods that employ several generations of selfing. The production of haploids and DHs provides a particularly attractive biotechnological tool, and the development of haploidy technology and protocols to produce homozygous plants has had a significant impact on agricultural systems. Nowadays, these biotechnologies represent an integral part of the breeding programmes of many agronomically important crops. There are several available methods to obtain haploids and DHs, of which in vitro anther or isolated microspore culture are the most effective and widely used. This review article deals with the current status of knowledge on the production of haploids and DHs through pollen embryogenesis and, in particular, anther culture.


Anther culture Doubled haploids Haploids Homozygosity Pollen embryogenesis 


  1. Achar PN (2002) A study of factors affecting embryo yields from anther culture of cabbage. Plant Cell Tissue Organ 69:183–188Google Scholar
  2. Aldemita RR, Zapata FJ (1991) Anther culture of rice: effects of radiation and media components on callus induction and plant regeneration. Cereal Res Commun 19:9–32Google Scholar
  3. Alemano L, Guiderdoni E (1994) Increased doubled haploid plant regeneration from rice (Oryza sativa L.) anther culture on colchicine-supplemented media. Plant Cell Rep 13:432–436Google Scholar
  4. Andersen SB (2005) Haploids in the improvement of woody species. In: Palmer CE, Keller WA, Kasha K (eds) Haploids in crop improvement II, vol 56. Springer, Heidelberg, pp 243–257Google Scholar
  5. Armstrong TA, Metz SG, Mascia PN (1987) Two regeneration system for the production of haploid plants from wheat anther culture. Plant Sci 51:231–237Google Scholar
  6. Ashok Kumar HG, Ravishankar BV, Murthy HN (2004) The influence of polyamines on androgenesis of Cucumis sativus L. Eur J Hortic Sci 69:201–205Google Scholar
  7. Atanassov A, Zagorska N, Boyadjiev P, Djilianov D (1995) In vitro production of haploid plants. World J Microbiol Biotechnol 11:400–408Google Scholar
  8. Bagni N, Tassoni A (2001) Biosynthesis, oxidation and conjugation of aliphatic polyamines in higher plants. Amino Acids 20:301–317PubMedGoogle Scholar
  9. Bajaj YPS (1990) In vitro production of haploids and their use in cell genetics and plant breeding. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, part I. Haploids in crop improvement, vol 12. Springer, Berlin, pp 1–44Google Scholar
  10. Bajaj YPS, Reinert J, Heberle E (1977) Factors enhancing in vitro production of haploid plants in anther and isolated microspore cultures. In: Gautheret R (ed) La culture des tissue et des cellules des végétaux. Mason, Paris, pp 47–58Google Scholar
  11. Ball ST, Zhou HP, Konzak CF (1993) Influence of 2, 4D, IAA and duration of callus induction in anther culture of spring wheat. Plant Sci 90:195–200Google Scholar
  12. Barnabas B, Phaler PL, Kovacs G (1991) Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat (Triticum aestivum L.). Theor Appl Genet 81:675–678Google Scholar
  13. Bhojwani SS, Razdan MK (1983) Plant tissue culture: theory and practice, vol 5. Elsevier, New YorkGoogle Scholar
  14. Bhojwani SS, Dunwell JM, Sunderland N (1973) Nucleic acid and protein contents of embryogenic tobacco pollen. J Exp Bot 24:863–871Google Scholar
  15. Binarovà P, Hause G, Cenklova V, Cordewener JHG, Van Lookeren Campagne MM (1997) A short-severe heat shock is required to induce embryogenesis in late bicellular pollen of Brassica napus L. Sex Plant Reprod 10:200–208Google Scholar
  16. Bjørnstad Å, Opsahl-Ferstad HG, Aasmo M (1989) Effects of donor plant environment and light during incubation of anther cultures of some spring wheat (Triticum aestivum L.) cultivars. Plant Cell Tissue Organ 17:27–37Google Scholar
  17. Blakeslee AF, Belling J, Farnham ME, Bergner AD (1922) A haploid mutant in the Jimson weed, Datura stramonium. Science 55:646–647PubMedGoogle Scholar
  18. Bohanec B (2003) Ploidy determination using flow cytometry. In: Maluszynski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 397–403Google Scholar
  19. Bonet FJ, Azhaid L, Olmedilla A (1998) Pollen embryogenesis: atavism or totipotency? Protoplasma 202:115–121Google Scholar
  20. Boutilier K, Offringa R, Sharma VK, Kieft H, Ouellet T, Zhang L, Hattori J, Liu CM, Van Lammeren AA, Miki BL, Custers JB, Van Lookeren Campagne MM (2002) Ectopic expression of BABY BOOM triggers a conversion from vegetative to embryonic growth. Plant Cell 14:1737–1749PubMedGoogle Scholar
  21. Bouvier L, Guérif P, Djulbic M, Durel CE, Chevreau E, Lespinasse Y (2002) Chromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers. Euphytica 123:255–262Google Scholar
  22. Broughton S (2008) Ovary co-culture improves embryo and green plant production in anther culture of Australian spring wheat (Triticum aestivum L.). Plant Cell Tissue Organ 95:185–195Google Scholar
  23. Castillo AM, Cistue L, Valles MP, Sanz L, Romagosa I, Molina-Cano JL (2001) Efficient production of androgenic doubled-haploid mutants in barley by the application of sodium azide to anther and microspore cultures. Plant Cell Rep 20:105–111Google Scholar
  24. Castillo AM, Cistue L, Valles MP, Soriano M (2009) Chromosome doubling in monocots. In: Touraev A, Forster B, Jain M (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 329–338Google Scholar
  25. Chen Z (1985) A study on induction of plants from Citrus pollen. Fruit Var J 39:44–50Google Scholar
  26. Chen CC, Kasha KJ, Marsolais A (1984) Segmentation patterns and mechanisms of genome multiplication in cultured microspores of barley. Can J Genet Cytol 26:475–483Google Scholar
  27. Chen QF, Wang CL, Lu YM, Shen M, Afza A, Duren MV, Brunner H (2001) Anther culture in connection with induced mutations for rice improvement. Euphytica 120:401–408Google Scholar
  28. Chiancone B, Tassoni A, Bagni N, Germanà MA (2006) Effect of polyamines on in vitro anther culture of Citrus clementina Hort. ex Tan. Plant Cell Tissue Organ 87:145–153Google Scholar
  29. Chu C (1978) The N6 medium and its applications to anther culture of cereal crops. In: Proc Symp Plant Tissue Culture. Science Press, Peking, pp 43–50Google Scholar
  30. Chupeau Y, Caboche M, Henry Y (1998) Androgenesis and haploid plants. INRA/Springer, BerlinGoogle Scholar
  31. Cistué L, Valles MP, Echavarri B, Sanz JM, Castillo AM (2003) Barley anther culture. In: Maluszynsky M, Kasha KJ, Forster BP, Szaejko I (eds) Doubled haploid production in crop plants. A manual. Kluwer/FAO-IAEA, Dordrecht/Vienna, pp 29–34Google Scholar
  32. Corduan G (1975) Regeneration of anther derived plants from anthers of Hyoscyamus niger. Planta (Berl) 127:27–36Google Scholar
  33. Deng XX, Deng ZA, Xiao SY, Zhang, WC (1992) Pollen derived plantlets from anther culture of Ichang papeda hybrids No. 14 and Trifoliate orange. In: Proc Int Soc Citriculture. Acireale, Italy, pp 190–192Google Scholar
  34. D’Amato F (1977) Cytogenetics of differentiation in tissue and cell cultures. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer, Berlin, pp 343–356Google Scholar
  35. Datta SK (2005) Androgenic haploids: factors controlling development and its application in crop improvement. Curr Sci 89:1870–1878Google Scholar
  36. Dunwell JM (1979) Anther culture in Nicotiana tabacum: the role of the culture vessel atmosphere in pollen embryo induction and growth. J Exp Bot 30:419–428Google Scholar
  37. Dunwell JM (1981) Stimulation of pollen embryo induction in tobacco by pretreatment of excised anthers in a water-saturated atmosphere. Plant Sci Lett 21:9–13Google Scholar
  38. Dunwell JM (1985) Embryogenesis from pollen in vitro. In: Zaitlin P, Day P, Hollaender A (eds) Biotechnology in plant science. Academic Press, Orlando, pp 49–76Google Scholar
  39. Dunwell JM (1986) Pollen, ovule and embryo culture, as tools in plant breeding. In: Withers LA, Alderson PG (eds) Plant tissue culture and its agricultural applications. Butterworths, London, pp 375–404Google Scholar
  40. Dunwell JM (2009) Patents and haploid plants. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 97–113Google Scholar
  41. Dunwell JM (2010) Haploids in flowering plants: origins and exploitation. Plant Biotechnol J 8:377–424PubMedGoogle Scholar
  42. Dunwell JM, Cornish M (1985) Influence of preculture variables on microspore embryo production in Brassica napus ssp. oleifera cv. Duplo. Ann Bot 56:281–289Google Scholar
  43. Dunwell JM, Sunderland N (1974a) Pollen ultrastructure in anther cultures of Nicotiana tabacum. I. Early stages of culture. J Exp Bot 25:352–361Google Scholar
  44. Dunwell JM, Sunderland N (1974b) Pollen ultrastructure in anther cultures of Nicotiana tabacum. II. Changes associated with embryogenesis. J Exp Bot 25:363–373Google Scholar
  45. Dunwell JM, Sunderland N (1975) Pollen ultrastructure in anther cultures of Nicotiana tabacum. III. The first sporophytic division. J Exp Bot 26:240–252Google Scholar
  46. Dunwell JM, Sunderland N (1976a) Pollen ultrastructure in anther cultures of Datura innoxia. I. Division of the presumptive vegetative cell. J Cell Sci 22:469–480PubMedGoogle Scholar
  47. Dunwell JM, Sunderland N (1976b) Pollen ultrastructure in anther cultures of Datura innoxia. II. The generative cell wall. J Cell Sci 22:481–492PubMedGoogle Scholar
  48. Dunwell JM, Sunderland N (1976c) Pollen ultrastructure in anther cultures of Datura innoxia. III. Incomplete microspore division. J Cell Sci 22:493–502PubMedGoogle Scholar
  49. Dunwell JM, Thurling N (1985) Role of sucrose in microspore embryo production in Brassica napus ssp. oleifera. J Exp Bot 36:1478–1491Google Scholar
  50. Dunwell JM, Cornish M, De Courcel AGL, Middlefell-Williams JE (1983) Induction and growth of ‘microspore-derived’ embryos of Brassica napus ssp. oleifera. J Exp Bot 34(12):1768–1778Google Scholar
  51. Evans DA, Sharp WR, Medina-Filho HP (1984) Somaclonal and gametoclonal variation. Am J Bot 71:759–774Google Scholar
  52. Ferrie AMR (2009) Current status of doubled haploids in medicinal plants. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 209–217Google Scholar
  53. Foroughi-Wehr B, Mix G (1976) In vitro responses of Hordeum vulgare L. anthers cultured from plants grown under different environments. Environ Exp Bot 19:303–309Google Scholar
  54. Foroughi-Wehr B, Friedt W, Wenzel G (1982) On the genetic improvement of androgenetic haploid formation in Hordeum vulgare L. Theor Appl Genet 62:246–248Google Scholar
  55. Forster BP, Thomas WTB (2005) Doubled haploids in genetics and plant breeding. Plant Breed Rev 25:57–88Google Scholar
  56. Forster BP, Herberle-Bors E, Kasha KJ, Touraev A (2007) The resurgence of haploids in higher plants. Trends Plant Sci 12(8):368–375PubMedGoogle Scholar
  57. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirement suspension cultures of soybean root cells. Exp Cell Res 50:151–158PubMedGoogle Scholar
  58. Germanà MA (1997) Haploidy in Citrus. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 5. Kluwer, Dordrecht, pp 195–217Google Scholar
  59. Germanà MA (2003) Haploids and doubled haploids in Citrus spp. In: Maluszynsky M, Kasha KJ, Forster BP, Szaejko I (eds) Doubled haploid production in crop plants. A manual. Kluwer/FAO-IAEA, Dordrecht/Vienna, pp 303–308Google Scholar
  60. Germanà MA (2005) Protocol of somatic embryogenesis from Citrus spp. anther culture. In Jain SM, Gupta PK (eds) Protocol of somatic embryogenesis-woody plants. Kluwer, Dordrecht, pp 191–207Google Scholar
  61. Germanà MA (2006) Doubled haploid production in fruit crops. Plant Cell Tissue Organ 86:131–146Google Scholar
  62. Germanà MA (2007) Haploidy. In: Khan I (ed) Citrus. Genetics, breeding and biotechnology. CABI, Wallingford, pp 167–196Google Scholar
  63. Germanà MA (2009) Haploid and doubled haploids in fruit trees. In: Touraev A, Forster B, Jain M (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 241–263Google Scholar
  64. Germanà MA, Chiancone B (2003) Improvement of the anther culture protocol in Citrus clementina Hort. ex Tan. Plant Cell Rep 22:181–187PubMedGoogle Scholar
  65. Germanà MA, Reforgiato G (1997) Haploid embryos regeneration from anther culture of ‘Mapo’ tangelo (Citrus deliciosa × C. paradisi). Adv Hortic Sci 11:147–152Google Scholar
  66. Germanà MA, Crescimanno FG, De Pasquale F, Wang YY (1991) Androgenesis in 5 cultivars of Citrus limon L. Burm. f. Acta Hortic 300:315–324Google Scholar
  67. Germanà MA, Wang YY, Barbagallo MG, Iannolino G, Crescimanno FG (1994) Recovery of haploid and diploid plantlets from anther culture of Citrus clementina Hort. ex Tan. and Citrus reticulata Blanco. J Hortic Sci 69(3):473–480Google Scholar
  68. Germanà MA, Crescimanno FG, Motisi A (2000a) Factors affecting androgenesis in Citrus clementina Hort. ex Tan. Adv Hortic Sci 14:43–51Google Scholar
  69. Germanà MA, Crescimanno FG, Reforgiato G, Russo MP (2000b) Preliminary characterization of several doubled haploids of Citrus clementina cv. Nules. In: Goren R, Goldschmidt EE (eds) Proc 1st Int Symp Citrus Biotechnology. Eilat, Israel, Acta Hortic 535:183–190Google Scholar
  70. Germanà MA, Chiancone B, Lain O, Testolin R (2005a) Anther culture in Citrus clementina: a way to regenerate tri-haploids. Aust J Agric Res 56:839–845Google Scholar
  71. Germanà MA, Chiancone B, Iaconia C, Muleo R (2005b) The effect of light quality on anther culture of Citrus clementina Hort. ex Tan. Acta Physiol Plant 27(4B):717–721Google Scholar
  72. Gervais C, Newcomb W, Simmonds DH (2000) Rearrangement of the actin filament and microtubule cytoskeleton during induction of microspore embryogenesis in Brasssica napus L. cv. Topas. Protoplasma 213:194–202Google Scholar
  73. Gonzalez-Melendi P, Ramırez 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 222:47–57PubMedGoogle Scholar
  74. Grewal RK, Lulsdorf M, Croser J, Ochatt S, Vandenberg A, Warkentin TD (2009) Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments. Plant Cell Rep 28:1289–1299PubMedGoogle Scholar
  75. Guha S, Maheshwari SC (1964) In vitro production of embryos from anthers of Datura. Nature 204:497–498Google Scholar
  76. Hause B, van Veenendaal WLH, Hause G, van Lammeren AAM (1994) Expression of polarity during early development of microsporederived and zygotic embryos of Brassica napus L. cv. Topas. Bot Acta 107:407–415Google Scholar
  77. Heberle-Bors E (1982) In vitro pollen embryogenesis in Nicotiana tabacum L. and its relation to pollen sterility, sex balance and floral induction of the pollen donor plants. Planta 156:396–401Google Scholar
  78. Heberle-Bors E (1983) Induction of embryogenic pollen grains and subsequent embryogenesis in Nicotiana tabacum L. by treatments of the pollen donor plants with feminizing agents. Physiol Plantarum 59:67–72Google Scholar
  79. Heberle-Bors E (1985) In vitro haploid formation from pollen: a critical review. Theor Appl Genet 71:361–374Google Scholar
  80. Heberle-Bors E (1989) Isolated pollen culture in tobacco: plant reproductive development in a nutshell. Sex Plant Reprod 2:1–10Google Scholar
  81. Heberle-Bors E, Reinert J (1981) Environmental control and evidence for predetermination of pollen embryogenesis in Nicotiana tabacum pollen. Protoplasma 109:249–255Google Scholar
  82. Hidaka T, Yamada Y, Shichijo T (1979) In vitro differentiation of haploid plants by anther culture in Poncirus trifoliata (L.) Raf. Jpn J Breed 29:248–254Google Scholar
  83. Hidaka T, Yamada Y, Shichijo T (1981) Plantlet formation from anthers of Citrus aurantium L.. Proc Int Soc Citriculture 1:153–155Google Scholar
  84. Ho KM, Jones GE (1980) Mingo barley. Can J Plant Sci 60:279–280Google Scholar
  85. Hochholdinger F, Hoecker N (2007) Towards the molecular basis of heterosis. Trends Plant Sci 12:427–432PubMedGoogle Scholar
  86. Hoekstra S, van Zijderveld MH, Louwerse JD, Heidekamp F, van der Mark F (1992) Anther and microspore culture of Hordeum vulgare L. cv Igri. Plant Sci 86:89–96Google Scholar
  87. Höfer M (1994) In vitro Androgenesis in apple: induction, regeneration and ploidy level. In: Schmidt H, Kellerhals M (eds) Progress in temperate fruit breeding. Kluwer, Dordrecht, pp 193–197Google Scholar
  88. Höfer M, Grafe C (2000) Preliminary evaluation of doubled haploid-material in apple. Acta Hortic 538:587–592Google Scholar
  89. Höfer M, Gomez A, Aguiriano E, Manzanera JA, Bueno MA (2002) Analysis of simple sequence repeat markers in homozygous lines in apple. Plant Breed 121:159–162Google Scholar
  90. Hosp J, Maraschin SF, Touraev A, Boutilier K (2007) Functional genomics of microspore embryogenesis. Euphytica 158:275–285Google Scholar
  91. Hu H, Yang HY (eds) (1986) Haploids in higher plants in vitro. China Academic Publishers/Springer, Beijing/BerlinGoogle Scholar
  92. Hu G, Liang GH, Wassom CE (1991) Chemical induction of apomictic seed formation in maize. Euphytica 56:97–105Google Scholar
  93. Huang B (1992) Genetic manipulation of microspores and microspore-derived embryos. In Vitro Cell Dev Biol 28:53–58Google Scholar
  94. Huang B (1996) Gametoclonal variation in crop improvement. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 2. Kluwer, Dordrecht, pp 73–91Google Scholar
  95. Immonen S, Anttila H (1996) Success in rye anther culture. Votr Pflanzenzuchtg 35:237–244Google Scholar
  96. Immonen S, Robinson J (2000) Stress treatments and ficoll for improving green plant regeneration in triticale anther culture. Plant Sci 150:77–84Google Scholar
  97. Jacquard C, Wojnarowiez G, Clément C (2003) Anther culture in barley. In: Maluszynsky M, Kasha KJ, Forster BP, Szaejko I (eds) Doubled haploid production in crop plants. A manual. Kluwer/FAO-IAEA, Dordrecht/Vienna, pp 21–27Google Scholar
  98. Jain SM, Sopory SK, Veilleux RE (eds) (1996–1997) In vitro haploid production in higher plants, vol 1–5. Kluwer, DordrechtGoogle Scholar
  99. Kadota M, Niimi Y (2004) Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai) by anther culture. Euphytica 138:141–147Google Scholar
  100. Kasha KJ (ed) (1974) Haploids in higher plants: advances and potential. The Office of Continuing Education, University of Guelph Press, GuelphGoogle Scholar
  101. Kasha KJ (2005) Chromosome doubling and recovery of doubled haploid plants. In: Palmer CE, Keller WA, Kasha KJ (eds) Haploids in crop improvement II, vol 56. Springer, Heidelberg, pp 123–152Google Scholar
  102. Kasha KJ, Kao KN (1970) High frequency haploid production in barley (Hordeum vulgare L.). Nature 225:874–876PubMedGoogle Scholar
  103. Kasha KJ, Maluszynsky M (2003) Production of doubled haploids in crop plants. An introduction. In: Maluszynsky M, Kasha KJ, Forster BP, Szaejko I (eds) Doubled haploid production in crop plants. A manual. Kluwer/FAO-IAEA, Dordrecht/Vienna, pp 1–4Google Scholar
  104. Kasha KJ, Hu TC, Oro R, Simion E, Shim YS (2001) Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley (Hordeum vulgare L.) microspores. J Exp Bot 52(359):1227–1238PubMedGoogle Scholar
  105. Keller WA, Stringham GR (1978) Production and utilization of microspore derived plants. In: Thorpe TA (ed) Frontiers of plant tissue culture. Calgary University Press, Calgary, pp 113–122Google Scholar
  106. Keller WA, Rajhathy T, Lacapra J (1975) In vitro production of plants from pollen in Brassica campestris. Can J Genet Cytol 17:655–666Google Scholar
  107. Keller WA, Armstrong KC, De La Roche AI (1983) The production and utilization of microspore-derived haploids in Brassica crops. In: Sen SK, Giles KL (eds) Plant cell culture in crop improvement. Plenum Press, New York, pp 169–183Google Scholar
  108. Kenis K, Keulemans J (2000) The use of microsatellites to investigate the homozygous status of apple plants obtained by anther culture and parthenogenesis in situ. Acta Hortic 538:581–585Google Scholar
  109. Khush GS, Virmani SS (1996) Haploids in plant breeding. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 1. Kluwer, Dordrecht, pp 11–33Google Scholar
  110. Kopecky D, Vagera J (2005) The use of mutagens to increase the efficiency of the androgenic progeny production in Solanum nigrum. Biol Plant 49:181–186Google Scholar
  111. Kott L (1998) Application of doubled haploid technology in breeding of oilseed Brassica napus. AgBiotech News Inf 10:69N–74NGoogle Scholar
  112. Krämer U, Pickering IJ, Prince RG, Raskin I, Salt DE (2000) Subcellular localization and speciation of Nickel in Hyperaccumulator and non-accumulator Thlaspi species. Plant Physiol 122:1343–1353PubMedGoogle Scholar
  113. Kumar A, Altabella T, Taylor MA, Tiburcio AF (1997) Recent advances in polyamine research. Trends in Plant Sci 2(4):124–130Google Scholar
  114. Kunzel G, Korzuna L, Meistera A (2000) Cytologically integrated physical restriction fragment length polymorphism maps for the barley genome based on translocation breakpoints. Genetics 154:397–412PubMedGoogle Scholar
  115. Kupper H, Lombi E, Zhao FJ, McGrath SP (2000) Cellular compartmentation of cadmium and zinc in relation to other elements in the hyperaccumulator Arabidopsis halleri. Planta 212:75–84PubMedGoogle Scholar
  116. Kyo M, Harada H (1986) Control of the developmental pathway of tobacco pollen in vitro. Planta 168:427–432Google Scholar
  117. Lazar MD, Schaeffer GW, Baenziger PS (1984) Cultivar and cultivar x environment effects on the development of callus and polyhaploid plants from anther cultures of wheat. Theor Appl Genet 67:273–277Google Scholar
  118. Lee LP, Hecht A (1975) Chloroplasts of monoploid and diploid Oenothera hookeri. Am J Bot 62:268–272Google Scholar
  119. Lee JH, Lee SY (2002) Selection of stable mutants from cultured rice anthers treated with ethyl methane sulfonic acid. Plant Cell Tissue Organ 71:165–171Google Scholar
  120. Letarte J, Simion E, Miner M, Kasha KJ (2006) Arabinogalactans and arabinogalactan-proteins induce embryogenesis in wheat (Triticum aestivum L.) microspore culture. Plant Cell Rep 24:691–698PubMedGoogle Scholar
  121. Levan A (1945) A haploid sugar beet after colchicine treatment. Hereditas 31:399–410PubMedGoogle Scholar
  122. Liang GH, Xu A, Tang H (1987) Direct generation of wheat haploids via anther culture. Crop Sci 27:336–339Google Scholar
  123. Magoon ML, Khanna KR (1963) Haploids. Caryologia 16:191–234Google Scholar
  124. Maheshwari SC, Rashid A, Tyagy AK (1982) Haploid from pollen grains-retrospect and prospect. Am J Bot 69:865–879Google Scholar
  125. Malik MR, Wang F, Dirpaul JM, Zhou N, Polowick PL, Ferrie AMR, Krochko JE (2007) Transcript profiling and identification of molecular markers for early microspore embryogenesis in Brassica napus. Plant Physiol 144:134–154PubMedGoogle Scholar
  126. Maluszynski M, Szarejko I, Barriga P, Balcerzyk A (2001) Heterosis in crop mutant crosses and production of high yielding lines using doubled haploid systems. Euphytica 120:387–398Google Scholar
  127. Maluszynski M, Kasha KJ, Forster BP, Szarejko I (eds) (2003a) Doubled haploid production in crop plants: a manual. Kluwer, DordrechtGoogle Scholar
  128. Maluszynski M, Kasha KJ, Szarejko I (2003b) Published double haploid protocols in plant species. In: Maluszynski M, Kasha KJ, Forster BP, Szarejko I (eds) Haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 309–335Google Scholar
  129. Maraschin SD, Lamers GEM, de Pater BS, Spaink HP, Wang M (2003) 14-3-3 isoforms and pattern formation during barley microspore embryogenesis. J Exp Bot 54:1033–1104Google Scholar
  130. Maraschin SF, de Priester W, Spaink HP, Wang M (2005a) Androgenic switch: an example of plant embryogenesis from the male gametophyte perspective. J Exp Bot 56(417):1711–1726PubMedGoogle Scholar
  131. Maraschin SF, Gaussand G, Olmedilla A, Pulido A, Lamers GEM, Korthout H, Spaink HP, Wang M (2005b) Programmed cell death during the transition from multicellular structures into globular embryos in barley androgenesis. Planta 221:459–470Google Scholar
  132. Martin B, Widholm JM (1996) Ploidy of small individual embryo-like structures from maize anther cultures treated with chromosome doubling agents and calli derived from them. Plant Cell Rep 15(10):781–785Google Scholar
  133. McCallum CM, Comai L, Greene EA, Henikoff S (2000) Targeted screening for induced mutations. Nat Biotechnol 18:455–457PubMedGoogle Scholar
  134. Misoo S, Yokota F, Matsubayashi M (1981) Effects of inoculation-ways of anthers on the pollen mitosis and plantlet formation in tobacco anther culture. Rep Soc Crop Sci Breed Kinki 26:44–48Google Scholar
  135. Mollers C, Iqbal MCM, Roblen G (1994) Efficient production of doubled haploid Brassica napus plants by colchicine treatment of microspores. Euphytica 75:95–104Google Scholar
  136. Morrison RA, Evans DA (1987) Gametoclonal variation. Plant Breed Rev 5:359–391Google Scholar
  137. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:473–497Google Scholar
  138. Narayanaswamy S, George L (1982) Anther culture. In: Johri BM (ed) Experimental embryology of vascular plants. Springer, Berlin, pp 79–103Google Scholar
  139. Nitsch C (1977) Culture of isolate microspore. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue and organ culture. Springer, Berlin, pp 268–278Google Scholar
  140. Nitsch C (1981) Production of isogenic lines: basic technical aspects of androgenesis. In: Thorpe TA (ed) Plant tissue culture methods and applications in agriculture. Academic Press, New York, pp 241–252Google Scholar
  141. Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163:85–87PubMedGoogle Scholar
  142. Nitsch C, Norreel B (1973) Effet d’un choc thermique sur le pouvoir embryogène du pollen de Datura innoxia cultivé dans l’anthère ou isolé de l’anthère. C R Acad Sci Paris 276D:303–306Google Scholar
  143. Nitzche W (1970) Herstellung haploider Pflanzen aus Festuca-Lolium Bastarden. Naturwissenschaft 57:199–200Google Scholar
  144. Obert B, Barnabás B (2004) Colchicine induced embryogenesis in maize. Plant Cell Tissue Organ 77:283–285Google Scholar
  145. Osolnik B, Bohanec B, Jelaska S (1993) Stimulation of androgenesis in white cabbage (Brassica oleracea var. capitata) anthers by low temperature and anther dissection. Plant Cell Tissue Organ 32:241–246Google Scholar
  146. Palmer CE, Keller WA, Kasha KJ (eds) (2005) Haploids in crop improvement II. Springer, Heidelberg, Germany, vol 56Google Scholar
  147. Pelletier G, Ilami M (1972) Les facteurs de l’androgenese in vitro chez Nicotiana tabacum. Z Pflanzenphysiol 68:97–114Google Scholar
  148. Perry JA, Wang TL, Welham TJ, Gardner S, Pike JM, Yoshida S, Parniske M (2003) A TILLING reverse genetics tool and a Web-accessible collection of mutants of the legume Lotus japonicus. Plant Physiol 131:866–871PubMedGoogle Scholar
  149. Petolino JF, Thompson SA (1987) Genetic analysis of anther culture response in maize. Theor Appl Genet 74:284–286Google Scholar
  150. Pintos B, Manzanera JA, Bueno MA (2007) Antimitotic agents increase the production of doubled-haploid embryos from cork oak anther culture. J Plant Physiol 164:1595–1604PubMedGoogle Scholar
  151. Powell W (1988) The influence of genotype and temperature pre-treatment on anther culture response in barley (Hordeum vulgare L.). Plant Cell Tissue Organ 12:291–297Google Scholar
  152. Powell W (1990) Environmental and genetic aspects of pollen embryogenesis. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, part I. Haploids in crop improvement, vol 12. Springer, Berlin, pp 44–65Google Scholar
  153. Pratap A, Gupta SK, Takahata Y (2009) Microsporogenesis and haploidy breeding. In: Gupta SK (ed) Biology and breeding of crucifers. CRC Press, Boca Raton, pp 293–307Google Scholar
  154. Pulido A, Bakos F, Castillo A, Vallés MP, Barnabas B, Olmedilla A (2005) Cytological and ultrastructural changes induced in anther and isolated-microspore cultures in barley: Fe deposits in isolated-microspore cultures. J Struct Biol 149:170–181PubMedGoogle Scholar
  155. Qin X, Rotino GL (1995) Chloroplast number in guard cells as ploidy indicator of in vitro-grown androgenic pepper plantlets. Plant Cell Tissue Organ 41:145–149Google Scholar
  156. Raghavan V (1990) From microspore to embryo: faces of the angiosperm pollen grain. In: Nijkamp HJJ, van der Plas LH, van Hartrigik J (eds) Progress in plant cellular and molecular biology. I.A.P.T.C. Kluwer, Dordrecht, pp 213–221Google Scholar
  157. Raghavan V (1997) Molecular embryology of flowering plants. Cambridge University Press, New YorkGoogle Scholar
  158. Rahman MH, Krishnaraj S, Thorpe TA (1995) Selection for salt tolerance in vitro using microspore-derived embryos of Brassica napus cv Topas, and the characterization of putative tolerant plants. In Vitro Cell Dev Biol Plant 31:116–121Google Scholar
  159. Rajyalakshmi K, Chowdhry CN, Maheshwari N, Maheshwari SC (1995) Anther culture response in some Indian wheat cultivars and the role of polyamines in induction of haploids. Phytomorphology 45:139–145Google Scholar
  160. Raquin C (1983) Utilization of different sugars of carbon sources for in vitro anther culture of Petunia. Z Pflanzenzucht 453–457Google Scholar
  161. Raquin C, Pilet V (1972) Production de plantes a partir d’antheres de Petuniaa cultiveés in vitro. Compt Rend D 274:1019–1022Google Scholar
  162. Reinert J, Bajaj YPS (1977) Anther culture: haploid production and its significance. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue and organ culture. Springer, Berlin, pp 251–267Google Scholar
  163. Reynolds TL (1997) Pollen embryogenesis. Plant Mol Biol 33:1–10PubMedGoogle Scholar
  164. Reynolds TL, Crawford RL (1997) Effects of light on the accumulation of abscisic acid and expression of an early cysteine-labeled metallothionein gene in microspores of Triticum aestivum during induced embryogenic development. Plant Cell Rep 16(7):458–463Google Scholar
  165. Rihova L, Tupy J (1999) Manipulation of division symmetry and developmental fate in cultures of potato microspores. Plant Cell Tissue Organ 59:135–145Google Scholar
  166. Rudolf K, Bohanec B, Hansen M (1999) Microspore culture of white cabbage, Brassica oleracea var. capitata L.: genetic improvement of non-responsive cultivars and effect of genome doubling agents. Plant Breed 118:237–241Google Scholar
  167. Sanders ME, Franzke CJ (1962) Somatic reduction of tetraploid sorghum to diploid mutants following colchicine treatment. Nature 196:696–698Google Scholar
  168. Sangwan RS, Sangwan-Norreel BS (1990) Anther and pollen culture. In: Bhajwari SS (ed) Plant tissue culture. Applications and limitations. Elsevier, Amsterdam, pp 220–241Google Scholar
  169. Sangwan-Norreel BS (1983) Male gametophyte nuclear DNA content evolution during androgenic induction in Datura innoxia. Z Pflanzenphysiol 111:47–54Google Scholar
  170. Sangwan-Norreel BS, Sangwan RS, Pare J (1986) Haploïdie et embryogenèse provoquée in vitro. Bull Society of Botany Fr 133. Actual Bot 4:7–39Google Scholar
  171. Seguí-Simarro JM (2010) Androgenesis revisited. Bot Rev 76:377–404Google Scholar
  172. Seguì-Simarro JM, Nuez F (2008a) How microspores transform into haploid embryos: changes associated with embryogenesis induction and microsporederived embryogenesis. Physiol Plant 134:1–12PubMedGoogle Scholar
  173. Seguì-Simarro JM, Nuez F (2008b) Pathways to doubled haploidy: chromosome doubling during androgenesis. Cytogenet Genome Res 120:358–369PubMedGoogle Scholar
  174. Shannon PRM, Nicholson AE, Dunwell JM, Davies DR (1985) Effect of anther orientation on microspore-callus production in barley (Hordeum vulgare). Plant Cell Tissue Organ 4:271–280Google Scholar
  175. Shariatpanahi ME, Bal U, Heberle-Bors E, Touraev A (2006) Stresses applied for the re-programming of plant microspores towards in vitro embryogenesis. Physiol Plant 127:519–534Google Scholar
  176. Sharma AK, Sharma A (1972) Chromosome techniques. Butterworths/University Park Press, London/BaltimoreGoogle Scholar
  177. Shim YS, Kasha KJ, Simion E, Letarte J (2006) The relationship between induction of embryogenesis and chromosome doubling in microspore cultures. Protoplasma 228:79–86PubMedGoogle Scholar
  178. Shull JK, Menzel MY (1977) A study of the reliability of synchrony in the development of pollen mother cells of Lilium longiflorum at the first meiotic prophase. Am J Bot 6 4:670–679Google Scholar
  179. Simantel GM, Ross JG (1964) Colchicine-induced somatic chromosome reduction in sorghum: IV. An induced haploid mutant. J Hered 55:3–5PubMedGoogle Scholar
  180. Skrzypek E, Czyczyło-Mysza I, Marcinska I, Wedzony M (2008) Prospects of androgenetic induction in Lupinus spp. Plant Cell Tissue Organ 94:131–137Google Scholar
  181. Smykal P (2000) Pollen embryogenesis—the stress mediated switch from gametophytic to sporophytic development. Current status and future prospects. Biol Plant 43:481–489Google Scholar
  182. Sopory SK, Maheshwari SC (1976) Development of pollen embryoids in anther culture of Datura innoxia. I. General observations and effects of physical factors. J Exp Bot 27:49–57Google Scholar
  183. Sopory S, Munshi M (1996) Anther culture. In: Mohan JM et al (eds) In vitro haploid production in higher plants, vol 1. Kluwer, Dordrecht, pp 145–176Google Scholar
  184. Srivastava P, Chaturvedi R (2008) In vitro androgenesis in tree species: an update and prospect for further research. Biotechnol Adv 26:482–491PubMedGoogle Scholar
  185. Stuart DA, Strickland SG, Walker KA (1987) Bioreactor production of alfalfa somatic embryos. Hortscience 22:800–809Google Scholar
  186. Sunderland N (1971) Anther culture: a progress report. Sci Prog (Oxford) 59:527–549Google Scholar
  187. Sunderland N (1974) Anther culture as means of haploid induction. In: Kasha KJ (ed) Haploids in higher plants. Advances and potential. University of Guelph Press, Guelph, pp 91–122Google Scholar
  188. Sunderland N (1978) Strategies in the improvement of yields in anther culture. In: Proc Symp Plant Tissue Culture. Science Press, Peking, pp 65–86Google Scholar
  189. Sunderland N, Dunwell JM (1972) Conditions for embryoid formation in Nicotiana pollen. Ann Rep John Innes Inst 62:60–61Google Scholar
  190. Sunderland N, Dunwell JM (1977) Anther and pollen culture. In: Street HE (ed) Plant tissue and cell culture. Oxford, Blackwell, pp 223–265Google Scholar
  191. Supena EDJ, Suharsono S, Jacobsen E, Custers JBM (2006) Successful development of a shed-microspore culture protocol for doubled haploid production in Indonesian hot pepper (Capsicum annuum L.). Plant Cell Rep 25:1–10PubMedGoogle Scholar
  192. Szarejko I, Forster BP (2006) Doubled haploidy and induced mutation. Euphytica 158:359–370Google Scholar
  193. Szarejko I, Forster BP (2007) Doubled haploidy and induced mutation. Euphytica 158(3):359–370Google Scholar
  194. Telmer CA, Newcom W, Simmonds DH (1992) Determination of developmental stage to obtain high frequencies of embryogenic microspores in Brassica napus. Physiol Plant 84:417–424Google Scholar
  195. Telmer CA, Newcomb W, Simmonds DH (1995) Cellular changes during heat shock induction and embryo development of cultured microspores of Brassica napus cv. Topas. Protoplasma 185:106–112Google Scholar
  196. Testillano PS, Coronado MJ, Seguì-Simarro JM, Domenech J, Gonzalez-Melendi P, Raska I, Risueno MC (2000) Defined nuclear changes accompany the reprogramming of the microspore to embryogenesis. J Struct Biol 129:223–232PubMedGoogle Scholar
  197. Thompson KF (1972) Oil-seed rape. In: Reports of the Plant Breeding Institute. Cambridge University Press, Cambridge, pp 94–96Google Scholar
  198. Tiainen T (1992) The role of ethylene and reducing agents on anther culture response of tetraploid potato (Solanum tuberosum L.). Plant Cell Rep 10:604–607Google Scholar
  199. Toojinda T, Baird E, Booth A, Broers L, Hayes P, Powell W, Thomas W, Vivar H, Young G (1998) Introgression of quantitative trait loci (QTLs) determining stripe rust resistance in barley: an example of marker-assisted development. Theor Appl Genet 96:123–131Google Scholar
  200. Touraev A, Ilham A, Vicente O, Heberle-Bors E (1996a) Stress induced microspore embryogenesis from tobacco microspores: an optimized system for molecular studies. Plant Cell Rep 15:561–565Google Scholar
  201. Touraev A, Indrianto A, Wratschko I, Vicente O, Heberle-Bors E (1996b) Efficient microspore embryogenesis in wheat (Triticum aestivum L.) induced by starvation at high temperatures. Sex Plant Reprod 9:209–215Google Scholar
  202. Touraev A, Vicente O, Heberle-Bors E (1997) Initiation of microspore embryogenesis by stress. Trends Plant Sci 2:297–302Google Scholar
  203. Touraev A, Pfosser M, Heberle-Bors E (2001) The microspore: a haploid multipurpose cell. Adv Bot Res 35:53–109Google Scholar
  204. Touraev A, Forster BP, Jain SM (eds) (2009) Advances in haploid production in higher plants. Springer, BerlinGoogle Scholar
  205. Tsay H-S (1981) Effects of nitrogen supply to donor plants on pollen embryogenesis in cultured tobacco anthers. J Agric Res China 30:5–13Google Scholar
  206. Turner J, Facciotti D (1990) High oleic acid Brassica napus from mutagenized microspores. In: McFerson JR, Kresovich S, Dwyer SG (eds) Proc 6th Crucifer Genet Workshop. Geneva, pp 24Google Scholar
  207. Tuvesson S, Dayteg C, Hagberg P, Manninen O, Tanhuanpa P, Tenhola-Roininen T, Kiviharju E, Weyen J, Forster J, Schondelmaier J, Lafferty J, Marn M, Fleck A (2006) Molecular markers and doubled haploids in European plant breeding programmes. Euphytica 158:305–312Google Scholar
  208. Vagera J, Novotny J, Ohnoutkova L (2004) Induced androgenesis in vitro in mutated populations of barley, Hordeum vulgare. Plant Cell Tissue Organ 77:55–61Google Scholar
  209. Vasil IK (1967) Physiology and cytology of anther development. Biol Rev 42:327–373PubMedGoogle Scholar
  210. Vasil IK (1973) Plants: haploid tissue cultures. In: Kruse PF Jr, Patterson MK Jr (eds) Tissue cultures methods and application. Academic Press, New York, pp 157–161Google Scholar
  211. Vasil IK (1980) Androgenic haploids. Int Rev Cytol Suppl 11A:195–223Google Scholar
  212. Veilleux RE (1994) Development of new cultivars via anther culture. Hortscience 29(11):1238–1241Google Scholar
  213. Verdoodt L, Van Haute A, Goderis IJ, De Witte K, Keulemans J, Broothaerts W (1998) Use of the multi-allelic self-incompatibility gene in apple to assess homozygosity in shoots obtained through haploid induction. Theor Appl Genet 96:294–300Google Scholar
  214. Wang M, van Bergen S, Van Duijn B (2000) Insights into a key developmental switch and its importance for efficient plant breeding. Plant Physiol 124:523–530PubMedGoogle Scholar
  215. Wang Z, Taramino G, Yong D, Liu G, Tingey SV, Miao G-L, Wang G-L (2001) Rice ESTs with disease-resistance gene or defense-response gene-like sequences mapped to regions containing major resistance genes or QTLs. Mol Gen Genome 265:302–310Google Scholar
  216. Wedzony M, Forster BP, Zur I, Golemiec E, Szechynska-Hebda M, Dubas, Gotebiowska G (2009) Progress in doubled haploid technology in higher plants. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Berlin, pp 1–34Google Scholar
  217. Wenzel G, Foroughi-Wehr B (1984) Anther culture of cereals and grasses. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants I. Academic Press, New York, pp 311–327Google Scholar
  218. Wenzel G, Hoffmann F, Thomas E (1977) Increased induction and chromosome doubling of androgenetic haploid rye. Theor Appl Genet 51:81–86Google Scholar
  219. Wenzel G, Frei U, Jahoor A, Graner A, Foroughghi-Wehr B (1995) Haploids—an integral part of applied and basic research. In: Terzi M et al (eds) Current issues in plant molecular and cellular biology. Kluwer, Dordrecht, pp 127–135Google Scholar
  220. Werner K, Friedt W, Ordon F (2007) Localisation and combination of resistance genes against soil-borne viruses of barley using doubled haploidy and molecular markers. Euphytica 158:323–329Google Scholar
  221. Wojnarowiez G, Jacquard C, Devaux P, Sagwan RS, Clement C (2002) Influence of copper sulphate on anther culture in barley (Hordeum vulgare L.). Plant Sci 162:843–847Google Scholar
  222. Wong RSC, Swanson E (1991) Genetic modification of canola oil: high oleic acid canola. In: Haberstroh C, Morris CE (eds) Fat and cholesterol reduced food. Gulf, Houston, pp 154–164Google Scholar
  223. Xu L, Najeeb U, Tang GX, Gu HH, Zhang GQ, He Y, Zhou WJ (2007) Haploid and doubled haploid technology. Adv Bot Res 45:181–216Google Scholar
  224. Yangn HY, Zhou C (1979) Experimental researches on the two pathways of pollen development in Oryza sativa. Acta Bot Sin 21:345–351Google Scholar
  225. Yeung EC, Rahman MH, Thorpe TA (1996) Comparative development of zygotic and microspore-derived embryos in Brassica napus L. cv. Topas. I. Histodifferentiation. Int J Plant Sci 157:27–39Google Scholar
  226. Yuan S, Liu Y, Fang Z, Yang L, Zhuang M, Zhang Y, Sun P (2009) Study on the relationship between the ploidy level of microspore-derived plants and the number of chloroplast in stomatal guard cells in Brassica oleracea. Agric Sci China 8:939–946Google Scholar
  227. Zaki MAM, Dickinson HG (1995) Modification of cell development in vitro: the effect of colchicine on anther and isolated microspore culture in Brassica napus. Plant Cell Tissue Organ 40:255–270Google Scholar
  228. Zhang YX, Lespinasse Y, Chevreau E (1990) Induction of haploidy in fruit trees. Acta Hort 280:293–304Google Scholar
  229. Zhao JP, Simmonds DH, Newcomb W (1996) Induction of embryogenesis with colchicine instead of heat in microspores of Brassica napus L. cv. Topas. Planta 198:433–439Google Scholar
  230. Ziauddin A, Ziauddin A, Simion EK, Kasha J (1990) Improved plant regeneration from shed microspore culture in barley (Hordeum vulgare L.). Plant Cell Rep 9:69–72Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Dipartimento S.EN.FI.MI.ZO, Facoltà di AgrariaUniversità degli Studi di PalermoPalermoItaly

Personalised recommendations