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利用瓶培养富集技术从草甘膦工厂排污口的污泥中筛选到一株高抗600mmol/L草甘膦的菌种,命名为ND-H;通过形态学观察、18S rDNA和5.8S rDNA-ITS的序列分析确定ND-H为串珠状赤霉(Gibberella moniliformis),其高抗草甘膦的特性尚未见报道;采取称干重法测定菌株随时间变化和不同培育条件下的生长量,表明其在液体培养基中的对数生长期为24～84h,最适生长温度为28℃,最适环境pH值为7.0;克隆了该菌的EPSPS基因,该基因受草甘膦诱导上调表达。研究结果将为培育转基因抗草甘膦作物提供新的抗性基因资源。
A flask culture-enriched technology was used to screen a strain of glyphosate resistant to 600 mmol / L glyphosate from the sludge at the sewage outfall of glyphosate factory and named as ND-H. By morphological observation, 18S rDNA and 5.8S rDNA The sequence analysis of -ITS confirmed that ND-H was Gibberella moniliformis and its glyphosate-resistant characteristics had not been reported yet. The growth of the strain under different culture conditions was measured by the method of dry weight, The results showed that the logarithmic growth phase in liquid medium was 24-84 h, the optimum growth temperature was 28 ℃ and the optimum pH was 7.0. The EPSPS gene of this strain was cloned and the gene was up-regulated by glyphosate . The results of this study will provide new resistance gene resources for cultivating transgenic glyphosate-resistant crops.