Studies on breeding methodology using isolated microspore culture and genetic transformation in cruciferous vegetables
2000
Sato, T. (National Research Inst. of Vegetables, Ornamental Plants and Tea, Ano, Mie (Japan))
Isolated microspores of Chinese cabbage (Brassica campestris var. pekinensis) belonging to the Kenshin group were incubated in modified N and N medium (NITSCH and NITSCH 1967) containing 10% sucrose in darkness at 33 degrees C for one day followed by culture at 25 degrees C. After 14 days of culture, microspores developed into embryos ranging from globular to cotyledonary stages. Plants were efficiently regenerated after transfer of embryos to 1/2 MS medium (MURASHIGE and SKOOG 1962) with 0.5 to 1.0% sucrose without plant growth regulators and with 0.8 to 1.5% agar. Self-compatible Brassica napus cv. 'Westar' was transformed with SLG, the S-locus glycoprotein gene that encodes the S-locus-specific glycoprotein (SLSG). I showed that the transgenic plants synthesized SLSG with the same apparent charge, molecular weight, and antigen properties as that produced by the corresponding self-incompatible strains from which the cloned SLG genes were isolated. When fused to the beta-glucuronidase (GUS) reporter gene, the 5' upstream regulatory region of SLG induced high level expression in the papillae of transgenic Brassica plants. Histochemical and fluorometric assays revealed that, in addition to the primary site of expression in the stigmatic papillae, the SLG-GUS fusion was also expressed in the transmitting tissues of the stigma, style, and ovary, and in anther
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