Institut de la recherche interdisciplinaire de Toulouse

Association de la recherche interdisciplinaire

              

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Brassica napus Haploid and Doubled Haploid Production and Cloning via Combined Microspore Embryogenesis and Somatic embryogenesis

X XuHan, H-C Jing, X-F Cheng, A. Iwanowska, H. Keift, JHW Bergervoet, SPC Groot, RJ Bino, AAM van Lammeren

(EU FP5 programe, cooperation with PCB and PRI, Wageningen University, the Netherlands. Closed). 

 

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An experimental morphogenesis research by means of cell biology, plant physiology and biochemistry. 

  • The first insight into in situ ploidy changes following development of microspore-derived embryos, plants, and secondary somatic embryos and their derived plants. 

  • The first successful combination of pollen- and somatic embryogenesis, to achieve one-step cloning of doubled haploid (DH) genotypes. 

  • The first insight into in situ ploidy changes following development of microspore-derived embryos, plants, and secondary somatic embryos and their derived plants. 

Main conclusions obtained: 

  • Diploidization occurs at the beginning of embryogenesis

  • Diploidization occurs in all tissues/organs if available

  • Diploidization occurs in all stages, and in similar speed

  • Diploidization leads 50% microspore embryos (pEMs) to DH or H-DH (chimerical) level 

  • Ploidy doubling stops in most cases at the DH level, does not pass tetraploidy (TH)

  • Somatic embryos can be induced in most pEM-derived plantlets that enables genotype cloning

  • Ploidy doubling also occurs in somatic embryogenesis

  • Somatic embryo cells at least show nuclear DNA at 1C to 8C levels 

  • Similarity of  ploidy  between somatic embryo (sEM) and its tissue of origin

  • Somatic embryos show an H - TH ploidy range

  • DH Somatic embryos are stabilized at DH

 

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Plate 1

Morphogenesis of Brassica napus Topas microspore embryogenesis and somatic embryogenesis. 

(Photos from Protoplasma 208/240-247, 1999)

 

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Plate 2 (left) and 3 (right)

Ploidy analyssis via flow-cytometric examinations of nuclear DNA of microspore- and/or pollen-derived embryos (pEMs), pEM-derived plantlets, somatic embryos (sEM), and plants.

Abbreviations:  H = haploid       DH = doubled haploid      TH = tetraploid   pEM = microspore embryo/embryogensis   sEM = somatic embryo/embryogenesis

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(Figures from Protoplasma 208/240-247, 1999)

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Related References

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