摘要：丛枝菌根能提高小麦对磷的吸收和利用，为培育磷高效的小麦新品种，本研究对丛枝菌根诱导的TaPT16基因进行功能研究。本试验以周麦26为实验材料，采用不同磷浓度及菌根处理周麦26，利用实时定量PCR（real-time quantitative PCR，qRT-PCR）技术了新的磷转运蛋白基因（命名为TaPT16）。并用几丁质和病原菌处理小麦，对不同处理的TaPT16的表达量进了分析。发现TaPT16基因可能参与病原菌与植物的互作。同时克隆了TaPT16基因，通过Gateway技术构建TaPT16基因的过表达载体，用农杆菌介导的方法将重组质粒转化磷利用低效的小麦品种京411，经PCR验证后获得了5个阳性植株。该研究为TaPT16基因的后期功能研究奠定了基础。

关键词：丛枝菌根；小麦磷转运蛋白TaPT16；实时定量PCR；Gateway技术

The research of a arbuscular mycorrhiza induced wheat phosphate transport protein gene TaPT16 

Abstract: Arbuscular mycorrhiza(AM) can enhance the ability of absorbtion and utilization of phosphorous in wheat .To cultivate novel wheat species of high phosphorous efficiently, , this research focused on the function of TaPT16,  of which the expression pattern was increased dramatically after inducing by arbuscular mycorrhiza. Zhoumai 26 was selected as material, which was treated with  different concerntatin of phosphorous and various AM fungi to analysis the expression pattern of TaPT16, a novel phosphate transporter gene.  Meanwhile, Zhoumai 26 was treated with chitin and pathogens to analysis TaPT16, the results revealed that TaPT16 might be involved in the   interactions between plant and pathogens. The complete CDS sequence of TaTP16 was cloned, and the recombinant overexpression vector of TaTP16 was established with the gateway technique. By agrobacterium mediated, we transformed the recombinant plasmid into the wheat–Jing411, which is inefficient in utilizing phosphorous. Five positive transgenic plants were acquired in the study, which laid the foundation of the function research of TaPT16. 

key words: arbuscular mycorrhiza; wheat phosphorous transport protein TaPT16; real-time quantitative PCR; Gateway technique

目    录

摘要 1

引言 1

1.材料与方法 3

1.1实验材料与仪器 3

1.1.1实验材料 3

1.1.2实验仪器 3

1.1.3实验药品及试剂 4

1.2材料培养和处理 4

1.2.1菌根及低磷处理 4

1.2.2几丁质处理 4

1.2.3禾顶囊壳小麦变种处理 5

1.2.4麦根腐平脐蠕孢处理 5

1.3 TaPT16序列的获得 5

1.4不同条件处理TaPT16基因表达量分析 5

1.5 TaPT16的克隆 5

1.6 TaPT16的过表达载体构建 6

1.6.1 BP反应 6

1.6.2 LR反应 6

1.7过表达TaPT16 的阳性植株的验证 6

2.结果与讨论 6

2.1 TaPT16的获得 6

2.2不同条件处理的小麦TaPT16表达量分析 7

2.2.1菌根及低磷处理对TaPT16表达量的影响