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  • 1
    Electronic Resource
    Electronic Resource
    Application of trifluralin to embryogenic microspore cultures to generate doubled haploid plants in Brassica napus (1995)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 95 (1995), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Microspore or anther culture has been used to produce desirable meiotic recombinants in numerous species. However, the utilization of these recombinants relies on inefficient genome doubling procedures to obtain fertile doubled haploid plants. This study presents a simple and rapid procedure to generate fertile doubled haploids in Brassica napus cv. Topas using trifluralin (α,α,α-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine), a plant specific microtubule inhibitor. The effects of trifluralin on microtubule depolymerization and chromosome doubling in embryogenic microspore cultures of B. napus were examined and compared with those of colchicine. Indirect immunofluorescence labeling of isolated microspores indicated that microtubules were depolymerized within 30 min of trifluralin treatment and after 3–8 h of colchicine treatment. The direct application of these microtubule inhibitors to microspore cultures resulted in the recovery of fertile doubled haploid plants. Continuous culture in the presence of colchicine, was more effective than 18-h treatments for fertile plant production but resulted in abnormal embryo formation and recalcitrant plant regeneration. The application of 1 or 10 μM trifluralin during the first 18 h of microspore culture was found to be the superior method for doubled haploid production. The embryos generated after trifluralin treatment developed normally, germinated readily and of the plants produced, close to 60% were fertile. The use of trifluralin to double chromosomes very early in microspore cultures is a simple process requiring minimal manipulation and should be very useful for genetic studies and breeding programs of B. napus and possibly other species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1995.tb00842.x
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  • 2
    Electronic Resource
    Electronic Resource
    Significance of preprophase bands of microtubules in the induction of microspore embryogenesis of Brassica napus (1999)
    Springer
    Planta 208 (1999), S. 383-391 
    Details
    ISSN: 1432-2048
    Keywords: Key words: Brassica (microspore embryogenesis) ; Cell wall ; Embryogenesis ; Microspore (division symmetry) ; Microtubule ; Preprophase band
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Microspores of Brassica napus L. cv. Topas, undergo embryogenesis when cultured at 32.5 °C for the first 18–24 h and then at 25 °C. The first division in heat-treated microspores is a symmetric division in contrast to the asymmetric division found after the first pollen mitosis in-planta or in microspores cultured continuously at 25 °C. This asymmetric division is unique in higher plants as it results in daughter cells separated by a non-consolidated wall. The cytoskeleton has an important role in such morphological changes. We examined microtubule (MT) organization during the first 24 h of heat induction in the embryogenic B. napus cv. Topas and the non-embryogenic B. napus breeding line 0025. Preprophase bands (PPBs) of MTs appeared in cv. Topas microspores in late uninucleate microspores and in prophase figures after 4–8 h of heat treatment. However, more than 60% of the PPBs were not continuous bands. In contrast, PPBs were never observed in pollen mitosis; MT strands radiated from the surface of the nuclear envelope throughout microspore maturation to the end of prophase of pollen mitosis I, during in-planta development and in microspores cultured at 25 °C. Following 24 h of heat treatment, over 95% of the microspores appeared to have divided symmetrically as indicated by the similar size of the daughter nuclei, but only 7–16% of the microspores eventually formed embryos. Discontinuous walls were observed in more than 50% of the divisions and it is probable that the discontinuous PPBs gave rise to such wall abnormalities which may then obstruct embryo development. Preprophase bands were not formed in heat-treated microspores of the non-embryogenic line 0025 and the ensuing divisions showed discontinuous walls. It is concluded that the appearance of PPBs in heat-induced microspores marks sporophytic development and that continuous PPBs are required for cell wall consolidation and embryogenesis. It follows that induced structures with two equally condensed nuclei, do not necessarily denote symmetric divisions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004250050573
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  • 3
    Electronic Resource
    Electronic Resource
    High frequency production of doubled haploid plants of Brassica napus cv. Topas derived from colchicine-induced microspore embryogenesis without heat shock (1996)
    Springer
    Plant cell reports 15 (1996), S. 668-671 
    Details
    ISSN: 1432-203X
    Keywords: Brassica napus ; Chromosome doubling ; Colchicine ; Doubled haploids ; Microspore embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This report describes a very high genome doubling efficiency of Brassica napus cv. Topas plants, derived from microspores induced to undergo embryogenesis with a colchicine treatment, without the use of a heat treatment. The plants showed normal growth and development, and 90% were fertile. In contrast, only 6% of the plants derived from heat-induced embryos were fertile diploids. All cytological analysis of the progeny of fertile plants showed 2n=38 chromosomes. These results show that colchicine can simultaneously induce microspore embryogenesis and double the ploidy level to produce doubled haploid plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231921
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  • 4
    Electronic Resource
    Electronic Resource
    Induction of embryogenesis with colchicine instead of heat in microspores ofBrassica napus L. cv. Topas (1996)
    Springer
    Planta 198 (1996), S. 433-439 
    Details
    ISSN: 1432-2048
    Keywords: Brassica napus ; Colchicine ; Heat shock ; Microspore embryogenesis ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Prior to this report, heat treatment (32.5°C, 24 h) was the method used to induce embryogenesis fromBrassica napus microspores. Continuous culture at 25°C results in pollen development. This study shows that colchicine alone, at the non-inductive temperature of 25°C, can induce embryogenesis, thus demonstrating that heat shock is not required for embryogenic induction inB. napus cv. Topas. Embryogenic frequencies of over 15% were obtained by culturing isolated microspores with 25 μM colchicine for 42 h at 25°C. The microspore developmental stages responsive to colchicine were unicellular vacuolate and late unicellular, somewhat earlier stages than the population responsive to heat induction. Other groups have reported that heat-shock proteins are essential to the induction of embryogenesis. The present study offers a method of embryogenic induction without the use of heat which will allow discrimination between the factors associated with response to heat shock and those involved with changing cell development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00620060
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  • 5
    Electronic Resource
    Electronic Resource
    High frequency production of doubled haploid plants of Brassica napus cv. Topas derived from colchicine-induced microspore embryogenesis without heat shock (1996)
    Springer
    Plant cell reports 15 (1996), S. 668-671 
    Details
    ISSN: 1432-203X
    Keywords: Keywords:Brassica napus– Chromosome doubling – Colchicine – Doubled haploids – Microspore embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. This report describes a very high genome doubling efficiency of Brassica napus cv. Topas plants, derived from microspores induced to undergo embryogenesis with a colchicine treatment, without the use of a heat treatment. The plants showed normal growth and development, and 90% were fertile. In contrast, only 6% of the plants derived from heat-induced embryos were fertile diploids. All cytological analysis of the progeny of fertile plants showed 2n=38 chromosomes. These results show that colchicine can simultaneously induce microspore embryogenesis and double the ploidy level to produce doubled haploid plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050095
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  • 6
    Electronic Resource
    Electronic Resource
    Cellular changes during heat shock induction and embryo development of cultured microspores ofBrassica napus cv. Topas (1995)
    Springer
    Protoplasma 185 (1995), S. 106-112 
    Details
    ISSN: 1615-6102
    Keywords: Brassica napus ; Embryogenesis ; Heat shock ; Induction ; Microspore embryogenesis ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Brassica napus cv. Topas microspores, isolated and cultured near the time of the first pollen mitosis and subjected to a heat treatment of 24 h, can be induced to develop into haploid embryos. This is a study of microspore structure during induction and embryo determination. Early during the 32.5 °C incubation period the nucleus moved away from the edge of the cell, and granules, 30 to 60 nm in diameter, appeared in the mitochondria and as a cluster in the cytoplasm. Cells divided symmetrically and at the end of the heat treatment, acquired the features of induced bicellular structures described previously. The features persisted as the cells divided randomly within the exine for 4–7 days following heat induction. Multicellular structures released from the exine underwent periclinal divisions resulting in protoderm differentiation of the globular embryo, thus determining embryo development. The cytoplasm of early heart-stage embryos contains abundant polyribosomes. Non-embryogenic development was indicated by large accumulations of starch and/or lipid and thickened cell walls or an unorganized pattern of cell division following release of the multicellular structures from the exine. Embryogenesis is discussed in terms of induction, embryo determination and development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01272758
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  • 7
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    Significance of preprophase bands of microtubules in the induction of microspore embryogenesis of Brassica napus (1999)
    Springer
    Details
    Publication Date: 1999-05-17
    Print ISSN: 0032-0935
    Electronic ISSN: 1432-2048
    Topics: Biology
    Published by Springer
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  • 8
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    Induction of embryogenesis with colchicine instead of heat in microspores ofBrassica napus L. cv. Topas (1996)
    Springer
    Details
    Publication Date: 1996-01-01
    Print ISSN: 0032-0935
    Electronic ISSN: 1432-2048
    Topics: Biology
    Published by Springer
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