DowElanco Biotechnology Laboratory, 9410 Zionsville Rd., P.O. Box 68955, 46268-1053, Indianapolis, IN, USA.
Plant Cell Rep. 1993 Dec;13(2):63-8. doi: 10.1007/BF00235291.
Transgenic haploid maize (Zea mays L.) plants were obtained from protoplasts isolated from microspore-derived cell suspension cultures. Protoplasts were electroporated in the presence of plasmid DNA containing the gus A and npt II genes encoding ß-glucuronidase (GUS) and neomycin phosphotransferase II (NPT II), respectively. Transformed calli were selected and continuously maintained on kanamycin containing medium. Stable transformation was confirmed by enzyme assays and DNA. analysis. Stably transformed tissue was transferred to regeneration medium and several plants were obtained. Most plants showed NPT II activity, and some also showed GUS activity. Chromosome examinations performed on representative plants showed that they were haploid. As expected, these plants were infertile.
通过微孢子体衍生细胞悬浮培养物中分离的原生质体，获得了转基因单倍体玉米（Zea mays L.）植株。在含有分别编码β-葡萄糖醛酸酶（GUS）和新霉素磷酸转移酶 II（NPT II）的 gus A 和 npt II 基因的质粒 DNA 的存在下，对原生质体进行电穿孔。转化的愈伤组织被选择并在含有卡那霉素的培养基上连续维持。通过酶分析和 DNA 分析证实了稳定的转化。将稳定转化的组织转移到再生培养基中，并获得了几个植株。大多数植株表现出 NPT II 活性，有些植株也表现出 GUS 活性。对代表性植株进行的染色体检查表明，它们是单倍体。正如预期的那样，这些植株不育。
