质粒DNA物理形态和其它因素对获得可育转基因小麦的影响

EFFECTS OF THE PHYSICAL CONFIGURATION OF PLASMID DNA AND OTHER FACTORS ON PRODUCTION OF FERTILE TRANSGENIC WHEAT PLANTS

本工作分析了不同形态质粒DNA和未成熟胚高渗透压处理对基因枪小麦转化体系的适用性。高渗处理对瞬间表达和转基因小麦的再生均有明显的促进作用。轰击之前对质粒DNA进行变性处理导致瞬间表达反应大幅度下降,但稳定转化频率(指从100个轰击未成熟胚得到的再生可育转基因植株数)与双链DNA相差不大。使用单链质粒DNA、线性双链质粒DNA和环状双链质粒DNA均可以得到转基因小麦植株。迄今已得到26个不同的转基因冬小麦株系和4个不同的转基因春小麦株系。这些转基因小麦大多数已产生种子,几个春小麦株系已获第二代种子。

The applicability of hyperosmotic treatment and different configurations of plasmid DNA for stable transformation of wheat mediated by particle bombardment was investigated. Hyperosmotic treatment increased the frequency of transient expression and had also a positive effect on stable transformation. Denaturation of plasmid DNA prior to bombardment led to dramatic reduction of transient expression. However, there were no marked differences between single-stranded and double-stranded DNA in stable transformation. Single-stranded plasmid DNA, double-stranded plasmid DNA in linear state and double-stranded plasmid DNA in circular state could all be used to produce transgenic wheat plants. A total of 26 independent transgenic plants of winter wheat genotype Florida and 4 independent transgenic plants of spring wheat genotype Veery were obtained. Most transgenic plants have set seeds. T2 seeds of some spring wheat transgenic plants have also been harvested.