转Bt基因棉对AM真菌磷转运基因表达及对菌根吸收、转运磷的影响

项目名称： 转Bt基因棉对AM真菌磷转运基因表达及对菌根吸收、转运磷的影响

项目编号： No.41271474

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 天文学、地球科学

项目作者： 陈秀华

作者单位： 华中农业大学

项目金额： 75万元

中文摘要： AM真菌与植物共生形成菌根，AM真菌磷转运基因所编码的高亲和性磷转运蛋白显著促进植物对磷的吸收，磷转运基因的表达受AM真菌与植物共生过程诱导，共生过程与特性对植物基因型和杀虫剂敏感。Bt基因导入植物不仅改变植物基因组成，还引起杀虫蛋白在土壤中积累，影响AM真菌与植物共生特性，对磷转运基因表达的影响值得关注。本项目拟采用Ri T-DNA棉花毛状根-AM真菌隔室双重培养与盆栽Bt棉方法，应用实时荧光定量RT-PCR、荧光原位RT-PCR与激光共聚焦显微镜等技术，研究Bt棉/Bt蛋白对AM真菌共生特性及对磷转运基因在菌丝与根内共生体中表达的影响；应用同位素标记与能量散射X线谱分析系统，研究磷在菌丝、（菌）根中的吸收、转运特征，探明磷转运基因表达水平与菌根体系磷的吸收、转运的关系，揭示Bt基因导入影响棉花-AM真菌共生系统磷吸收的分子机制。研究结果对阐明Bt棉对AM真菌共生效应的影响具有重要意义。

中文关键词： 转Bt基因棉；丛枝菌根真菌；共生；共生效应；基因表达

英文摘要： The high-affinity phosphate (P) transporter of arbuscular mycorrhizal (AM) fungi can significantly improve the uptake of P by the host plants. The gene expression of P transporter is induced by the symbiotic processes with plant roots. However, the symbiotic relationship is sensitive to the plant gene structures and insecides. Transgenic Bacillus thuringiensis (Bt) plants not only have the different gene structures with the wide type, but also release insecticidal proteins to the soil, which has the effect on the symbiotic relationship of AM fungi with their hosts. Thus the effect on gene expression of P transporter is highly worthy of attention. This project plans to study the effect of transgenic Bt cotton and Bt insecticidal protein on the symbiotic character and P transporter gene expression of AM fungi. During the research, Ri T-DNA-transformed cotton roots inoculated by AM fungi spores will be grown in Petri plates with two compartments. Bt insecticidal protein will be added to the culture medium. The Bt and non-Bt cotton will be planted in pots, inoculated by AM fungi or not. Quantitative real-time PCR analyses, direct fluorescent in situ RT-PCR and confocal laser scanning microscopy will be used to examine phosphate transporter gene expression profiles in extraradical myceliun and mycorrhizal roots.33P a

英文关键词： transgenic Bacillus thuringiensis (Bt) cotton；AM fungi；colonization；symbiotic effect；gene expression