Resistance identification of bivalent fungi-resistant genes transformed soybean to Phytophthora sojae

Soybean is one of the most important sources of edible oil and proteins in the world. However, it suffers from many kinds of fungal diseases which is a major limiting factor in soybean production. The fungal disease can be effectively controlled by breeding plant cultivars with genetic transformation. In this study, the resistance to Phytophthora sojae of five bivalent transgenic soybean lines was identified using the hypocotyls inoculation technique. The lines were the T2 of the transgenic soybean which were transformed with kidney bean chitinase gene and barley ribosome inactivating protein gene, and were positive by Southern Blot analysis. The resistance difference was studied through comparing the death percentage of transgenic soybean with the control. The results showed that four lines were more resistant to P sojae, whereas other one had no significant difference in comparison with the control. These transgenic soybean lines with enhanced resistance to P sojae will be useful in soybean resistance breeding.

转几丁质酶和核糖体失活蛋白双价基因大豆的获得与抗病性鉴定

以东北地区4个主栽大豆品种为试材,通过农杆菌介导法,将2个抗真菌病基因,即几丁质酶基因和核糖体失活蛋白基因构建在同一植物表达载体上,对受体材料进行遗传转化,获得了经Southern检测同时整合双价基因的T0代转基因大豆植株,转化频率为0.5%。将T0代转基因大豆植株扩繁至T2代,并进行了大豆疫霉根腐病和大豆灰斑病的抗性检测,4个株系的抗性均比对照有明显提高。分子生物学验证及RT-PCR检测表明,2个外源基因在4个株系中均进行了转录表达。

大豆抗疫霉根腐病的蛋白组研究

利用双向电泳及MALDI-TOF-MS技术分析绥农10号真叶接种疫霉菌1号生理小种后的蛋白质组变化。在抗病品种绥农10号叶片中共获得19个差异表达蛋白点,其中有12个上调表达,6个下调表达,1个特异表达(仅在接种后出现)。利用生物质谱分析8个上调表达点、1个下调表达点和1个特异表达点,最终鉴定得到8个有注释功能的蛋白,根据功能可分为4类,第1类为参与新陈代谢的蛋白,包括二磷酸核酮糖羧化酶的大亚基及前体、琥珀酰-辅酶A;第2类为参与信号传导的蛋白,包括激酶受体类蛋白、氧化还原酶和半胱氨酸氧化还原酶;第3类为参与细胞内物质运输的蛋白,包括衣壳蛋白的zeta-3亚基;第4类为转录因子,是参与茉莉酸介导的F-box蛋白。这些蛋白可为进一步研究大豆抗病机制奠定基础。