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为明确腊梅硫酯酶基因(Cp FATB)在苜蓿中的表达特点及生理功能,获得苜蓿抗寒育种的基因资源。本实验通过根癌农杆菌介导将Cp FATB转入‘龙牧806’、‘WL319HQ’和‘肇东’3个苜蓿品种中,对其后代进行抗寒性鉴定。结果表明,3个苜蓿品种转Cp FATB基因SP3代(第三代基因修饰材料)种子发芽率、简化活力指数均高于各自对照,其中T‘龙牧806’最高,达到87.79%、208.19;转基因植株叶绿素a、叶绿素b及叶绿素总量显著低于其各自对照苜蓿品种,根系可溶性糖、可溶性蛋白、游离脯氨酸含量显著高于其各自对照苜蓿品种;转Cp FATB基因植株高度和分枝数与其各自对照差异不显著,而越冬率显著高于其各自对照品种,T‘龙牧806’越冬率最高,达到96.60%。转Cp FATB基因促进‘龙牧806’、‘WL319HQ’、‘肇东’苜蓿SP3代种子萌发;抑制苜蓿叶片叶绿素合成。越冬前促进根系可溶性糖、可溶性蛋白、游离脯氨酸含量积累,增强POD、SOD活性,提高了翌年越冬率,说明外源Cp FATB基因有助于苜蓿抗寒性的提高。
In order to clarify the expression characteristics and physiological functions of Cp FATB gene in alfalfa, the genetic resources of alfalfa cold-resistant breeding were obtained. In this experiment, Cp FATB mediated by Agrobacterium tumefaciens was transferred into three alfalfa varieties ’Longmu 806’, ’WL319HQ’ and ’Zhaodong’, and their cold resistance was identified. The results showed that the seed germination rate and the simplified viability index of three transgenic alfalfa varieties with CpGYB gene SP3 (third-generation genetically modified material) were higher than their respective controls, of which T’Longmu 806 ’was the highest (87.79%, 208.19; The total chlorophyll a, chlorophyll b and total chlorophyll of the plants were significantly lower than those of their respective control alfalfa cultivars. The content of soluble sugar, soluble protein and free proline in roots were significantly higher than that of their respective control alfalfa cultivars. There was no significant difference with their respective control, while the overwintering rate was significantly higher than their respective control varieties, T ’Longmu 806’ the highest overwintering rate, reaching 96.60%. Transfection with CpG FATB gene promoted seed germination of SP3 seedlings of ’Longmu 806’, ’WL319HQ’ and ’Zhaodong’ alfalfa and inhibited chlorophyll synthesis of alfalfa leaves. Before winter, the contents of soluble sugar, soluble protein and free proline in roots were increased, POD and SOD activities were increased, and the overwintering rate in the following year was increased, indicating that the exogenous Cp FATB gene was helpful for the improvement of cold resistance of alfalfa.