[Objective] The study aimed at evaluating the uncertainty in measuring the construct-specific fragments of genetically modified maize MON863 by real time quantitative PCR.[Method] The content of construct-specific fra...[Objective] The study aimed at evaluating the uncertainty in measuring the construct-specific fragments of genetically modified maize MON863 by real time quantitative PCR.[Method] The content of construct-specific fragments in MON863 samples was determined by real time quantitative PCR,and then the uncertainty of measurement result was evaluated according to the sources of uncertainty like the PCR system,the data processing and the micropipette.[Result] Type A evaluation of uncertainty（uA） in the measurement was 1.7×10^-2;Type B evaluation of uncertainty（uB） was 9.0×10^-4;the combined standard uncertainty（uC） was 1.7×10^-2;the expanded uncertainty（U95） was 0.036 and the finally measured result was 1.08%±0.036.[Conclusion] The main uncertainty of the result measured by real time quantitative PCR came from the randomizing effect in the experimental process.展开更多
[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quant...[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quantitative PCR detection method for genetically modified maize event NK603 was developed using primers and Taqman probe designed according to the flanking sequence of event NK603, which was then adopted to detect the samples containing 2% NK603 stand- ard （with uncertain quantity of 10% ）. [ Results ] The slope of standard curve ranged between -3.6 and -3.1, and the correlation coefficient was higher than 0. 99. The amplification efficiency of this method reached 100.2%, fallen between 90% and 110%. The detected quantity of the experimental sample was 1.9%, closer to the true quantity （2%）. [ Conclusion] This quantitative PCR detection method for genetically modified maize event NK603 is very precise and can be a- dopted in routine testing analysis.展开更多
The main advantage of digital PCR(dPCR) is that it facilitates absolute quantification of the target without reference to the standard/calibration curve.Crystal droplet dPCR has a three-color staining detection functi...The main advantage of digital PCR(dPCR) is that it facilitates absolute quantification of the target without reference to the standard/calibration curve.Crystal droplet dPCR has a three-color staining detection function,which enables multiplex PCR reaction.In this study,this technique was used to establish triple dPCR detection for the genetically modified soybean MON87708 × MON89788 with stacked traits.Specific absolute quantitative detection was accomplished for the genomic DNA extracted from the homogenized seeds of GM stack MON87708× MON89788 soybean.Our results can serve as a reference for the absolute quantitative detection of stacked events of genetically modified crops.展开更多
Unintended effects of genetic modification on chemical composition of Bt maize leaf litter may have impacts on its decomposition. In most agricultural systems in South Africa, maize litter is either left on the soil s...Unintended effects of genetic modification on chemical composition of Bt maize leaf litter may have impacts on its decomposition. In most agricultural systems in South Africa, maize litter is either left on the soil surface or incorporated into the soil during tillage. A litterbag experiment, using leaf litter of three maize hybrids （DKC80-12B, DKC80-10 and DKC6-125）, was carried out at the University of Fort Hare Research Farm, South Africa, to determine the effects of genetic modification on decomposition of maize leaf litter when left on the soil surface under field conditions between July and November, the normal fallow period, in 2008. Another litterbag experiment was conducted at the University of Fort Hare Research Farm and Zanyokwe Irrigation Scheme, South Africa, using leaf ~itter of two maize hybrids genetically modified with the erylAb gene （MONS10）, DKC75-15B and PAN6Q-3OSB, and their corresponding near isolines, CRN3505 and PAN6Q-121. The degradation of CrylAb protein in the litter, both surface-applied and soil-incorporated, was also investigated. Decomposition of Bt maize litter was similar to that of non-Bt maize litter both when applied on the surface and when incorporated into soil. Soil-incorporated litter, as well as its CrylAb protein, decomposed faster than that applied on the surface. The leaf litter C：N ratios of PAN6Q-308B and PAN6Q-121 were similar throughout the study, whereas those of DKC75-15B and CRN3505 declined by similar amounts during a 12-week period. These findings suggested that decomposition of leaf litter of Bt maize, with the MON810 event, was not affected by maize genetic modification, and that the CrylAb protein broke down together with plant leaf litter during the winter fallow regardless of whether the litter was applied on the soil surface or incorporated into soil.展开更多
基金Supported by the Fund from Sichuan Academy of Agricultural Sciences for Distinguished Young Scholars (2009QNJJ-037)a Grantfor Adventive Species Monitoring from Ministry of Agriculture~~
文摘[Objective] The study aimed at evaluating the uncertainty in measuring the construct-specific fragments of genetically modified maize MON863 by real time quantitative PCR.[Method] The content of construct-specific fragments in MON863 samples was determined by real time quantitative PCR,and then the uncertainty of measurement result was evaluated according to the sources of uncertainty like the PCR system,the data processing and the micropipette.[Result] Type A evaluation of uncertainty（uA） in the measurement was 1.7×10^-2;Type B evaluation of uncertainty（uB） was 9.0×10^-4;the combined standard uncertainty（uC） was 1.7×10^-2;the expanded uncertainty（U95） was 0.036 and the finally measured result was 1.08%±0.036.[Conclusion] The main uncertainty of the result measured by real time quantitative PCR came from the randomizing effect in the experimental process.
基金Supported by Youth Science and Technology Program of Sichuan Academy of Agricultural Science(2009QNJJ-037)Program for Monitoring Invasive Species of Ministry of Agriculture
文摘[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quantitative PCR detection method for genetically modified maize event NK603 was developed using primers and Taqman probe designed according to the flanking sequence of event NK603, which was then adopted to detect the samples containing 2% NK603 stand- ard （with uncertain quantity of 10% ）. [ Results ] The slope of standard curve ranged between -3.6 and -3.1, and the correlation coefficient was higher than 0. 99. The amplification efficiency of this method reached 100.2%, fallen between 90% and 110%. The detected quantity of the experimental sample was 1.9%, closer to the true quantity （2%）. [ Conclusion] This quantitative PCR detection method for genetically modified maize event NK603 is very precise and can be a- dopted in routine testing analysis.
文摘The main advantage of digital PCR(dPCR) is that it facilitates absolute quantification of the target without reference to the standard/calibration curve.Crystal droplet dPCR has a three-color staining detection function,which enables multiplex PCR reaction.In this study,this technique was used to establish triple dPCR detection for the genetically modified soybean MON87708 × MON89788 with stacked traits.Specific absolute quantitative detection was accomplished for the genomic DNA extracted from the homogenized seeds of GM stack MON87708× MON89788 soybean.Our results can serve as a reference for the absolute quantitative detection of stacked events of genetically modified crops.
基金Supported by the National Research Foundation of South Africa(NRF)the Govan Mbeki Research and Development Center(GMRDC)of the University of Fort Hare(No.GUN62299)
文摘Unintended effects of genetic modification on chemical composition of Bt maize leaf litter may have impacts on its decomposition. In most agricultural systems in South Africa, maize litter is either left on the soil surface or incorporated into the soil during tillage. A litterbag experiment, using leaf litter of three maize hybrids （DKC80-12B, DKC80-10 and DKC6-125）, was carried out at the University of Fort Hare Research Farm, South Africa, to determine the effects of genetic modification on decomposition of maize leaf litter when left on the soil surface under field conditions between July and November, the normal fallow period, in 2008. Another litterbag experiment was conducted at the University of Fort Hare Research Farm and Zanyokwe Irrigation Scheme, South Africa, using leaf ~itter of two maize hybrids genetically modified with the erylAb gene （MONS10）, DKC75-15B and PAN6Q-3OSB, and their corresponding near isolines, CRN3505 and PAN6Q-121. The degradation of CrylAb protein in the litter, both surface-applied and soil-incorporated, was also investigated. Decomposition of Bt maize litter was similar to that of non-Bt maize litter both when applied on the surface and when incorporated into soil. Soil-incorporated litter, as well as its CrylAb protein, decomposed faster than that applied on the surface. The leaf litter C：N ratios of PAN6Q-308B and PAN6Q-121 were similar throughout the study, whereas those of DKC75-15B and CRN3505 declined by similar amounts during a 12-week period. These findings suggested that decomposition of leaf litter of Bt maize, with the MON810 event, was not affected by maize genetic modification, and that the CrylAb protein broke down together with plant leaf litter during the winter fallow regardless of whether the litter was applied on the soil surface or incorporated into soil.
