目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
1材料与方法3
1.1材料 3
1.2方法 3
1.2.1构建载体3
1.2.2植物材料培养及叶片预处理 4
1.2.3钨弹制备4
1.2.4基因枪轰击转化5
1.2.5接种观察 5
1.2.6统计方法 5
2结果与分析5
2.1载体构建5
2.2 瞬时表达体系效率检测6
2.3 G1和G2单倍型的抗白粉病功能检测6
3讨论 8
致谢9
参考文献9
图21 G1和G2载体构建5
图22 小麦叶片细胞中GFP和GUS的表达 6
图23 典型的抗病互作细胞和感病互作细胞 6
图24 CK、G1、G2经白粉菌侵染后的平均吸器指数 8
表21 对照CK、G1、G2经白粉病菌侵染后观察到的吸器指数7
小麦抗白粉基因类似物TmRGA3的功能单倍型鉴定
摘 要
小麦是世界范围内广泛种植的谷类作物，在我国为仅次于水稻的第二大粮食作物，但其产量受到包括白粉病在内的多种病害的威胁。培育抗性品种是防治小麦白粉病最为有效安全的方法，而发掘有效抗源、鉴定抗性功能基因并加以利用是保证抗病育种持久性和稳定性的基础。鉴定抗白粉病基因功能常用的方法有稳定转化，叶片瞬时表达和病毒诱导的基因沉默技术。瞬时表达是在靶细胞内暂时高效表达目的基因的一种技术。与稳定转化相比，瞬时表达技术不需要将靶基因整合到基因组中，通过靶基因瞬时表达来鉴定其功能，可达到快速筛选基因的目的。鉴于其具有时间短，效率高等优点，在筛选抗白粉病基因的研究中得到了广泛应用。
实验前期从一粒小麦TA2033中鉴定到一个抗病基因TmRGA3，遗传转化实验表明，该基因能够提供白粉病抗性。通过对多份一粒小麦种质进行TmRGA3同源基因的克隆，又鉴定出12种单倍型。本实验构建了其中2种单倍型G1和G2的强启动子表达载体，利用基因枪介导的瞬时转化技术，将这2 *51今日免费论文网|www.51jrft.com +Q: @351916072@ 
种单倍型表达载体质粒轰击至植物表皮细胞，通过接种白粉病菌后统计吸器指数，分析其是否为抗白粉病功能单倍型。结果表明，G1单倍型具有明显的抗白粉病功能。
IDENTIFICATION OF FUNCTIONAL HAPLOTYPES OF WHEAT POWDERY MILDEW RESISTANCE GENE ANALOG TmRGA3
ABSTRACT
Wheat is a cereal crop widely planted worldwide, and it is the second largest food crop after rice in China, but its output is threatened by various diseases including powdery mildew. Cultivating resistant varieties is the most effective and safe method to control wheat powdery mildew, and discovering effective resistance sources, identifying resistance functional genes and using them are the basis for ensuring the durability and stability of diseaseresistant breeding.Common methods for identifying the function of powdery mildew resistance genes include stable transformation, transient leaf expression and virusinduced gene silencing techniques.Transient expression is a technique for temporarily expressing target genes efficiently in target cells. In contrast to stable transformation, transient expression techniques do not require the integration of target genes into the genome, and the identification of their function through transient expression of target genes can achieve the purpose of rapid gene screening.In view of its short time and high efficiency, it has been widely used in the research of screening powdery mildew resistance genes.
In the early stage of the experiment, a disease resistance gene TmRGA3 was identified from a wheat TA2033. Genetic transformation experiments showed that the gene could provide powdery mildew resistance.Twelve haplotypes were identified by cloning TmRGA3 homologous genes in multiple wheat germplasms.In this experiment, two strong haplotype G1 and G2 promoter expression vectors were constructed. Using gene gunmediated transient transformation technology, these two haplotype expression vector plasmids were bombarded into plant epidermal cells and inoculated with powdery mildew After the statistical suction index, analyze whether it is a haplotype of powdery mildew resistance.The results show that the G1 haplotype has obvious resistance to powdery mildew.

原文链接：http://www.jxszl.com/nongxue/zwbh/607461.html