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作者	单位
杜娟，吴季荣，俞明正，王燕霞，徐剑宏，史建荣	（1.南京农业大学植物保护学院植物病理学系，农业部作物病虫害监测与防控重点开放实验室，江苏南京 210095；2.江苏省农业科学院食品质量安全与检测研究所/江苏省转基因安全评价公共服务中心，江苏南京 210014）

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中文摘要:

荧光假单胞菌（Psdeuomnoda fluoerncnet）是一种土壤中十分常见的细菌，广泛应用于生物防治，其数量变化能较准确地反映土壤环境的变化，因此在转基因小麦环境安全评价研究中可以将荧光假单胞菌的数量变化作为检测转基因小麦种植对整个根际土壤环境影响的指标之一。本研究选取荧光假单胞菌的蛋白编码基因gyrB作为靶标基因，设计特异性较好的引物，利用该引物建立了基于实时荧光定量PCR（RFQ PCR）方法的转基因小麦根际土壤荧光假单胞菌数量的检测技术体系。结果显示，定量标准曲线相关系数达0.99以上，且呈现单一熔解曲线峰值。应用该体系对种植于河南新乡及江苏六合的转 TaDREB4基因抗旱小麦（简称TB4）及其受体小麦济麦19根际土壤荧光假单胞菌进行检测，结果表明，各时期转基因小麦和非转基因小麦各生育期根际土壤中荧光假单胞菌的拷贝数无显著差异。说明TB4种植对根际土壤中的荧光假单胞菌数量无显著影响。

英文摘要:

Psdeuomnoda fluoerncnet is one of the most popular bacterial in soil, and applied for biological pest control. The change of its amount and structure can also reflect the environmental change of soil. So it is often being applied as a biological indicator of the soil ecological security when performing risk assessment for transgenic plant. In this study, a method that based on real time flororescent quantitative PCR (RFQ PCR) was established for detecting the amount of P. fluorescens and was used for risk assessment of transgenic wheat. The gene gyrB were chosen as the target and the primers were designed for detecting the amount of P.fluorescens with RFQ PCR. With this method, the amount of the P.fluorescens in rhizospheric soil of TaDREB4 transgenic wheat (TB4) and its wild type Jimai 19 (JM19) was detected at different growth stage. The results showed that the correlation coefficient of the quantitative standard curve was above 0.99 and the melting peak was single. The testing result showed that there was no significant difference for quality of P.fluorescens in rhizospheric soils between TB4 and JM19 all over the growth stages. The result indicated that TB4 had no significant impact on the quality of P. fluorescens and this method could be used for risk assessment of transgenic plant.

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