Baumgarte Susanne, Tebbe Christoph C
Institut für Agrarökologie, Bundesforschungsanstalt für Landwirtschaft (FAL), Bundesallee 50, 38116 Braunschweig, Germany.
Mol Ecol. 2005 Jul;14(8):2539-51. doi: 10.1111/j.1365-294X.2005.02592.x.
Field studies were done to assess how much of the transgenic, insecticidal protein, Cry1Ab, encoded by a truncated cry1Ab gene from Bacillus thuringiensis (Bt), was released from Bt-maize MON810 into soil and whether bacterial communities inhabiting the rhizosphere of MON810 maize were different from those of the rhizosphere of nontransgenic maize cultivars. Bacterial community structure was investigated by SSCP (single-strand conformation polymorphism) of PCR-amplified 16S rRNA genes from community DNA. Using an improved extraction and detection protocol based on a commercially available ELISA, it was possible to detect Cry1Ab protein extracted from soils to a threshold concentration of 0.07 ng/g soil. From 100 ng of purified Cry1Ab protein added per gram of soil, only an average of 37% was extractable. At both field sites investigated, the amount of Cry1Ab protein in bulk soil of MON810 field plots was always lower than in the rhizosphere, the latter ranging from 0.1 to 10 ng/g soil. Immunoreactive Cry1Ab protein was also detected at 0.21 ng/g bulk soil 7 months after harvesting, i.e. in April of the following year. At this time, however, higher values were found in residues of leaves (21 ng/g) and of roots (183 ng/g), the latter corresponding to 12% of the Cry1Ab protein present in intact roots. A sampling 2 months later indicated further degradation of the protein. Despite the detection of Cry1Ab protein in the rhizosphere of MON810 maize, the bacterial community structure was less affected by the Cry1Ab protein than by other environmental factors, i.e. the age of the plants or field heterogeneities. The persistence of Cry1Ab protein emphasizes the importance of considering post-harvest effects on nontarget organisms.
开展了田间研究,以评估苏云金芽孢杆菌(Bt)截短的cry1Ab基因编码的转基因杀虫蛋白Cry1Ab从Bt玉米MON810释放到土壤中的量,以及MON810玉米根际的细菌群落与非转基因玉米品种根际的细菌群落是否存在差异。通过对群落DNA中PCR扩增的16S rRNA基因进行单链构象多态性(SSCP)分析,研究细菌群落结构。使用基于市售酶联免疫吸附测定(ELISA)的改进提取和检测方案,能够检测从土壤中提取的Cry1Ab蛋白,阈值浓度为0.07 ng/g土壤。每克土壤添加100 ng纯化的Cry1Ab蛋白后,平均只有37%可被提取。在所调查的两个田间地点,MON810田间地块的大块土壤中Cry1Ab蛋白的含量始终低于根际,后者的含量范围为0.1至10 ng/g土壤。收获7个月后,即次年4月,在大块土壤中也检测到了免疫反应性Cry1Ab蛋白,含量为0.21 ng/g。然而,此时在叶片残留物(21 ng/g)和根残留物(183 ng/g)中发现了更高的值,后者相当于完整根中Cry1Ab蛋白含量的12%。两个月后的一次采样表明该蛋白进一步降解。尽管在MON810玉米根际检测到了Cry1Ab蛋白,但细菌群落结构受Cry1Ab蛋白的影响小于受其他环境因素的影响,即植物年龄或田间异质性。Cry1Ab蛋白的持久性强调了考虑收获后对非靶标生物影响的重要性。