Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers.

In this work, some of the different in vitro developmental pathways into which tomato microspores or microsporocytes can be deviated experimentally were explored. The two principal ones are direct embryogenesis from isolated microspores and callus formation from meiocyte-containing anthers. By means of light and electron microscopy, the process of early embryogenesis from isolated microspores and the disruption of normal meiotic development and change of developmental fate towards callus proliferation, morphogenesis, and plant regeneration have been shown. From microspores isolated at the vacuolate stage, embryos can be directly induced, thus avoiding non-androgenic products. In contrast, several different morphogenic events can be triggered in cultures of microsporocyte-containing anthers under adequate conditions, including indirect embryogenesis, adventitious organogenesis, and plant regeneration. Both callus and regenerated plants may be haploid, diploid, and mostly mixoploid. The results demonstrate that both gametophytic and sporophytic calli occur in cultured tomato anthers, and point to an in vitro-induced disturbance of cytokinesis and subsequent fusion of daughter nuclei as a putative cause for mixoploidy and genome doubling during both tetrad compartmentalization and callus proliferation. The potential implications of the different alternative pathways are discussed in the context of their application to the production of doubled-haploid plants in tomato, which is still very poorly developed.

[1]  T. Varghese,et al.  Production of embryoids and calli from isolated microspores of tomato (Lycopersicon esculentumMill.) in liquid media , 1986, Biologia Plantarum.

[2]  Gulshan,et al.  Studies on anther cultures of tomato —Lycopersicon esculentumMill , 1981, Biologia Plantarum.

[3]  D. Verma,et al.  Cell division control in plants , 2008 .

[4]  E. Simion,et al.  Isolated microspore culture of wheat (Triticum aestivum L.) in a defined media I. Effects of pretreatment, isolation methods, and hormones , 1995, In Vitro Cellular & Developmental Biology - Plant.

[5]  J. M. Seguí-Simarro,et al.  Plant Cytokinesis - Insights Gained from Electron Tomography Studies , 2007 .

[6]  K. Mishiba,et al.  Ploidy distribution in the explant tissue and the calluses induced during the initial stage of internode segment culture of Asparagus officinalis L. , 2007, In Vitro Cellular & Developmental Biology - Plant.

[7]  E. Heberle‐Bors,et al.  Stresses applied for the re‐programming of plant microspores towards in vitro embryogenesis , 2006 .

[8]  J. M. Seguí-Simarro,et al.  Mechanisms of cytokinesis in flowering plants: new pieces for an old puzzle. , 2006 .

[9]  A. Jaime,et al.  Floriculture, ornamental and plant biotechnology : advances and topical issues , 2006 .

[10]  A. Meister,et al.  Colchicine-induced polyploidization depends on tubulin polymerization in c-metaphase cells , 2006, Protoplasma.

[11]  C. Ramírez,et al.  Three dimensional confocal and electron microscopy imaging define the dynamics and mechanisms of diploidisation at early stages of barley microspore-derived embryogenesis , 2005, Planta.

[12]  J. Reinert,et al.  Induction of haploid tobacco plants from isolated pollen , 1975, Protoplasma.

[13]  V. Mathur,et al.  Chromosomal Instability in Callus Culture of Pisum sativum , 2004, Plant Cell, Tissue and Organ Culture.

[14]  C. Dumas,et al.  Spontaneous chromosome doubling results from nuclear fusion during in vitro maize induced microspore embryogenesis , 2004, Chromosoma.

[15]  J. M. Seguí-Simarro,et al.  Electron Tomographic Analysis of Somatic Cell Plate Formation in Meristematic Cells of Arabidopsis Preserved by High-Pressure Freezing , 2004, The Plant Cell Online.

[16]  W. Zhou,et al.  Efficient production of doubled haploid plants by immediate colchicine treatment of isolated microspores in winter Brassica napus , 2002, Plant Growth Regulation.

[17]  V. Sotirova,et al.  Induced androgenesis in tomato (Lycopersicon esculentum Mill.). III. Characterization of the regenerants , 1998, Plant Cell Reports.

[18]  W. Newcomb,et al.  Induction of embryogenesis with colchicine instead of heat in microspores ofBrassica napus L. cv. Topas , 1996, Planta.

[19]  N. Batty,et al.  Ethylene precursors and antagonists increase embryogenesis of Hordeum vulgare L. anther culture , 1994, Plant Cell Reports.

[20]  E. Guiderdoni,et al.  Increased doubled haploid plant regeneration from rice (Oryza sativa L.) anthers cultured on colchicine-supplemented media , 1994, Plant Cell Reports.

[21]  J. Dunwell,et al.  Barley anther culture: Pretreatment on mannitol stimulates production of microspore-derived embryos , 1990, Plant Cell, Tissue and Organ Culture.

[22]  G. Ladizinsky,et al.  Lycopersicon esculentum: Trifoliate plants recovered from anther cultures of heterozygous tftf plants , 1984, Plant Cell Reports.

[23]  J. Dunwell,et al.  The role of nuclear fusion in pollen embryogenesis of Datura innoxia Mill. , 1974, Planta.

[24]  P. Gresshoff,et al.  Development and differentiation of haploid Lycopersicon esculentum (tomato) , 1972, Planta.

[25]  W. Zhou,et al.  Increasing embryogenesis and doubling efficiency by immediate colchicine treatment of isolated microspores in spring Brassica napus , 2004, Euphytica.

[26]  S. Jha,et al.  Induction of mitosis in polytene nuclei and hormonal effect on nuclear changes during callus initiation in diploid Urginea indica Kunth. (liliaceae) , 2004, Genetica.

[27]  H. Dickinson,et al.  Microspore-derived embryos in Brassica: the significance of division symmetry in pollen mitosis I to embryogenic development , 2004, Sexual Plant Reproduction.

[28]  L. Staehelin,et al.  Electron tomographic analysis of post-meiotic cytokinesis during pollen development in Arabidopsis thaliana , 2004, Planta.

[29]  M. Maluszynski,et al.  Doubled haploid production in crop plants : a manual , 2003 .

[30]  W. Thomas,et al.  Doubled haploids in breeding , 2003 .

[31]  I. Szarejko,et al.  Doubled Haploid Production in Crop Plants , 2003, Springer Netherlands.

[32]  Y. Shim,et al.  Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley (Hordeum vulgare L.) microspores. , 2001, Journal of experimental botany.

[33]  K. Mishiba,et al.  Increasing ploidy level in cell suspension cultures of Doritaenopsis by exogenous application of 2,4-dichlorophenoxyacetic acid. , 2001, Physiologia plantarum.

[34]  L. Shtereva,et al.  Callus induction and plant regeneration via anther culture in mutant tomato (Lycopersicon esculentum Mill.) lines with anther abnormalities , 2001 .

[35]  E. Heberle‐Bors,et al.  The microspore: A haploid multipurpose cell , 2001 .

[36]  G. Karlov,et al.  [Morphometric and cytogenetic characteristics of haploid tomato plants]. , 2000, Genetika.

[37]  Kenji Kato,et al.  Efficient callus induction and shoot regeneration by anther culture in male sterile mutants of tomato (Lycopersicon esculentum Mill. cv. First) , 1999 .

[38]  K. Kasha,et al.  A cytological study of pretreatments used to improve isolated microspore cultures of wheat (Triticum aestivum L.) cv. Chris , 1999 .

[39]  M. Guerra,et al.  Callus induction and plant regeneration of tomato (Lycopersicon esculentum cv. IPA 5) via anther culture , 1999 .

[40]  L. Shtereva,et al.  Induced androgenesis in tomato (Lycopersicon esculentum Mill). II. Factors affecting induction of androgenesis , 1998, Plant Cell Reports.

[41]  B. Dimitrov,et al.  Induced androgenesis in tomato (Lycopersicon esculentum Mill.) I. Influence of genotype on androgenetic ability , 1998, Plant Cell Reports.

[42]  B. Campion,et al.  An improved protocol for androgenesis in cauliflowers (Brassica oleracea var. botrytis) , 1997 .

[43]  M. Guedira,et al.  Role of temperature as a triggering signal for organogenesis or somatic embryogenesis in wounded leaves of chicory cultured in vitro , 1994 .

[44]  P. Gahan,et al.  Plant growth regulator-induced xylogenesis in cotyledons of Solanum aviculare , 1994 .

[45]  T. Bailey,et al.  Microspore developmental stage and anther length influence the induction of tomato anther callus , 1992 .

[46]  W. Summers,et al.  Dark-light treatments influence induction of tomato anther callus , 1991 .

[47]  S. Mantell,et al.  Early Cellular Events During Direct Somatic Embryogenesis in Cotyledon Explants of Solanum aviculare Forst. , 1991 .

[48]  W. Summers,et al.  Tomato Anther Callus Production: Solidifing Agent and Concentration Influence Induction of Callus , 1990 .

[49]  F. J. Zapata,et al.  Callus formation and plant regeneration in isolated pollen culture of rice (Oryza sativa L. cv. Taipei 309) , 1988 .

[50]  V. Sawhney,et al.  Ultrastructure of the shoot apex of tomato (Lycopersicon esculentum) , 1985 .

[51]  B. Juniper,et al.  The Ultrastructure of Pollen Grain Development in Lycopersicum Peruvianum , 1984 .

[52]  R. Lichter.  Induction of Haploid Plants From Isolated Pollen of Brassica napus , 1982 .

[53]  Z. Xu,et al.  Culture of Barley Anthers in Conditioned Media , 1981 .

[54]  M. Cappadocia,et al.  Plant regeneration from in vitro cultures of anthers and stem internodes in an interspecific hybrid, lycopersicon esculentum L. X L. peruvianum mill. and cytogenetic analysis of the regenerated plants , 1980 .

[55]  D. Zamir,et al.  Anther culture of male-sterile tomato (Lycopersicon esculentum mill.) mutants , 1980 .

[56]  W. R. Sharp,et al.  Haploid Plantlets and Callus from Immature Pollen Grains of Nicotiana and Lycopersicon , 1971 .

[57]  S. Maheshwari,et al.  In vitro Production of Embryos from Anthers of Datura , 1964, Nature.

[58]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .