利用反义RNA技术分析春化相关基因VER17在冬小麦花发育过程中的功能

Functional Analysis of Vernalization-related Gene VER17 in Flower Development Using Antisense RNA Strategy in Winter Wheat

为了研究小麦春化相关基因VER17的功能,应 用反义RNA技术,将VER17基因的反义片段构建到载 体pBI121上,通过花粉管通道法获取转基因小麦。对 T0代转基因植株GUS染色以及PCR等分子鉴定,得到 14株含反义VER17基因片段的阳性转基因植株。对T0 代和T1代的表型观察结果显示,VER17反义转基因植 株开花时间延迟,并且穗的顶部和基部小花出现明显 的退化。表明春化相关基因VER17在小麦发育过程中 可能起到促进植物开花以及穗顶端和基部花发育的作 用,减少小花退化,同时对雄蕊的发育也有影响。

To understand the function of vernalization-related gene VER17 in winter wheat (Triticum aestivum L. cv. Jingdong No.1), an antisense RNA strategy was used. The antisense VER17 with a vector pBI121 was constructed (Fig.1) and transformed into winter wheat by using the pollen-tube-pathway method. Fourteen independent transgenic plants transformed with antisense VER17 and five control transformants transformed with pBI121 blank vector were obtained (Table 1) and confirmed by GUS his-tochemical assay (Fig.2) and PCR-Southern blot analysis (Fig.3). Phenotypes of T0 and T1 transgenic plants showed that the plants of the antisense VER17 transgenic lines degenerated top or basal spikelets (Fig.4, Fig.5) and had delayed flowering time (Fig.6), which suggested that the VER17 gene functions in accelerating flowering and the development of the top or basal spikelets and the stamen which were impacted by vernalization treatment.