[Objective]This study aimed to establish an effective detection technology for rapidly identifying Bt transgenic sugarcane,promptly removing non-Bt transgenic plants and increasing breeding efficiency of Bt transgenic...[Objective]This study aimed to establish an effective detection technology for rapidly identifying Bt transgenic sugarcane,promptly removing non-Bt transgenic plants and increasing breeding efficiency of Bt transgenic plants,thereby improving breeding efficiency of insect-resistant sugarcane cultivars in China.[Method]Approximately 1 mg of root tips and other materials were crushed,placed in the bottom of PCR tubes,added successively with solution I and mineral oil,headed at 95 ℃ for 15 min,added with solution II,and finally added with solution III containing GBt primer for PCR. After separated by using electrophoresis on agarose gel,amplification products were observed and photographed under a gel imaging system. [Result]Various materials treated with different concentrations of solution led to different amplification results. Specifically,the appropriate concentration of NaOH in solution I was 0. 1- 0. 2 mol /L; the appropriate pH value of solution II was 2. No bands were amplified from mature leaves,tender leaves and old roots of Bt transgenic sugarcane,while root tips successfully amplified 545bp target bands. The established method was used to identify and analyze 63 hybrid seedlings of YT 91- 976 × SBR216,results showed that a total of 17 samples exhibited 545 bp bands,accounting for approximately 27% of the total number of hybrid seedlings,indicating that these 17 seedlings were Bt transgenic sugarcane plants. [Conclusion]The rapid detection method established in this study is conducive to identifying Bt transgenic sugarcane and improving breeding efficiency of insect-resistant sugarcane lines,which provides basis for screening of other transgenic materials.展开更多
Chemical reactions and fate of the toxins of Bacillus thuringiensis(Bt) in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants.In this study,the effect of io...Chemical reactions and fate of the toxins of Bacillus thuringiensis(Bt) in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants.In this study,the effect of ionic strength(0-1 000 mmol kg 1) adjusted by NaCl or CaCl 2 on adsorption of Bt toxin by a lateritic red soil,a paddy soil and these soils after chemical removal of organic-bound or free Fe and Al oxides,as well as by pure minerals(goethite,hematite and gibbsite) which are widespread in these soils,were studied.The results indicated that when the supporting electrolyte was NaCl,the adsorption of Bt toxin by the lateritic red soil and paddy soil increased rapidly until the ionic strength reached 250 mmol kg 1 and then gradually slowed down with the increase of ionic strength;while in case the supporting electrolyte was CaCl 2,the adsorption of Bt toxin enhanced significantly at low ionic strength(< 10 mmol kg 1) and then decreased as the ionic strength increased.The adsorption of Bt toxin by the tested minerals and soils after the removal of organic-bound or free Fe and Al oxides also increased with increasing ionic strength controlled by NaCl.Removing organic-bound Fe and Al oxides obviously increased the adsorption of Bt toxin in the tested soils.Differently,removing free Fe and Al oxides increased the Bt adsorption by the paddy soil,but decreased the adsorption by the lateritic red soil.The study indicated that the varieties of ionic strength and the presence of Fe and Al oxides affected the adsorption of Bt toxin by the soils,which would contribute to the further understanding of the fate of Bt toxin in the soil environment and provide references for the ecological risk assessment of transgenic Bt plants.展开更多
基金Supported by Modern Agricultural Industry Technology System (CARS-2004B)
文摘[Objective]This study aimed to establish an effective detection technology for rapidly identifying Bt transgenic sugarcane,promptly removing non-Bt transgenic plants and increasing breeding efficiency of Bt transgenic plants,thereby improving breeding efficiency of insect-resistant sugarcane cultivars in China.[Method]Approximately 1 mg of root tips and other materials were crushed,placed in the bottom of PCR tubes,added successively with solution I and mineral oil,headed at 95 ℃ for 15 min,added with solution II,and finally added with solution III containing GBt primer for PCR. After separated by using electrophoresis on agarose gel,amplification products were observed and photographed under a gel imaging system. [Result]Various materials treated with different concentrations of solution led to different amplification results. Specifically,the appropriate concentration of NaOH in solution I was 0. 1- 0. 2 mol /L; the appropriate pH value of solution II was 2. No bands were amplified from mature leaves,tender leaves and old roots of Bt transgenic sugarcane,while root tips successfully amplified 545bp target bands. The established method was used to identify and analyze 63 hybrid seedlings of YT 91- 976 × SBR216,results showed that a total of 17 samples exhibited 545 bp bands,accounting for approximately 27% of the total number of hybrid seedlings,indicating that these 17 seedlings were Bt transgenic sugarcane plants. [Conclusion]The rapid detection method established in this study is conducive to identifying Bt transgenic sugarcane and improving breeding efficiency of insect-resistant sugarcane lines,which provides basis for screening of other transgenic materials.
基金Supported by the National Natural Science Foundation of China (Nos. 41001140 and 40671087)
文摘Chemical reactions and fate of the toxins of Bacillus thuringiensis(Bt) in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants.In this study,the effect of ionic strength(0-1 000 mmol kg 1) adjusted by NaCl or CaCl 2 on adsorption of Bt toxin by a lateritic red soil,a paddy soil and these soils after chemical removal of organic-bound or free Fe and Al oxides,as well as by pure minerals(goethite,hematite and gibbsite) which are widespread in these soils,were studied.The results indicated that when the supporting electrolyte was NaCl,the adsorption of Bt toxin by the lateritic red soil and paddy soil increased rapidly until the ionic strength reached 250 mmol kg 1 and then gradually slowed down with the increase of ionic strength;while in case the supporting electrolyte was CaCl 2,the adsorption of Bt toxin enhanced significantly at low ionic strength(< 10 mmol kg 1) and then decreased as the ionic strength increased.The adsorption of Bt toxin by the tested minerals and soils after the removal of organic-bound or free Fe and Al oxides also increased with increasing ionic strength controlled by NaCl.Removing organic-bound Fe and Al oxides obviously increased the adsorption of Bt toxin in the tested soils.Differently,removing free Fe and Al oxides increased the Bt adsorption by the paddy soil,but decreased the adsorption by the lateritic red soil.The study indicated that the varieties of ionic strength and the presence of Fe and Al oxides affected the adsorption of Bt toxin by the soils,which would contribute to the further understanding of the fate of Bt toxin in the soil environment and provide references for the ecological risk assessment of transgenic Bt plants.
