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Field studies on the environmental fate of the Cry1Ab Bt-toxin produced by transgenic maize (MON810) and its effect on bacterial communities in the maize rhizosphere

机译:转基因玉米(MON810)产生的Cry1Ab Bt毒素的环境命运及其对玉米根际细菌群落的影响的田间研究

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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.
机译:进行了田间研究,以评估由苏云金芽胞杆菌(Bt)的一个被截断的cry1Ab基因编码的转基因杀虫蛋白Cry1Ab是从Bt玉米MON810中释放到土壤中的,以及是否存在于居住在MON810玉米根际中的细菌群落与非转基因玉米品种的根际不同。通过社区DNA的PCR扩增的16S rRNA基因的SSCP(单链构象多态性)研究了细菌群落结构。使用基于市售ELISA的改进的提取和检测方案,可以检测从土壤中提取的Cry1Ab蛋白达到阈值浓度0.07 ng / g。从每克土壤中添加的100 ng纯化Cry1Ab蛋白中,平均只能提取到37%。在两个实地调查中,MON810田块大块土壤中Cry1Ab蛋白的含量始终低于根际,后者在0.1至10 ng / g土壤之间。收获后7个月(即次年4月)还以0.21 ng / g的散装土壤检测到了免疫反应性Cry1Ab蛋白。然而,此时,在叶片(21 ng / g)和根(183 ng / g)的残留物中发现了更高的值,后者对应于完整根中存在的Cry1Ab蛋白的12%。 2个月后的采样表明该蛋白质进一步降解。尽管在MON810玉米的根际中检测到Cry1Ab蛋白,但细菌群落结构受Cry1Ab蛋白的影响要小于其他环境因素(即植物的年龄或田间异质性)的影响。 Cry1Ab蛋白的持久性强调了考虑收获后对非目标生物的影响的重要性。

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