A PCR-ELISA method for detecting the glyphosate resistant transgenic soybean was established and optimized. The results showed that the key parameters of PCR-ELISA were as follows： the concentration of digoxin tag pr...A PCR-ELISA method for detecting the glyphosate resistant transgenic soybean was established and optimized. The results showed that the key parameters of PCR-ELISA were as follows： the concentration of digoxin tag probe was 0.5 μmol · L^-1, the time of hybridization reaction was 15 min and the chromogenic reaction should last for 30 min. The sensitivity and the repeatability of our PCR-ELISA method were evaluated, and the results showed that it could be detected when the concentration of DNA template from transgenic soybean samples was 0.01% or higher, and the coefficient of variation of this method was less than 5% in our research condition. These results suggested that PCR-ELISA method establishment in this study had good repeatability and high precision for detecting the transgenic soybean samples.展开更多
Competitions of light,nutrient and water between transgenic glyphosate resistant soybean and weed were evaluated with different herbicides(glyphosate,acetochlor,and the mixture of fomesafen and quizalofop-p-ethyl)in f...Competitions of light,nutrient and water between transgenic glyphosate resistant soybean and weed were evaluated with different herbicides(glyphosate,acetochlor,and the mixture of fomesafen and quizalofop-p-ethyl)in filed,which displayed that the application of the mixture of fomesafen and quizalofop-p-ethyl on soybeans had the best anti-herbicidal effect while glyphosate had the weakest effect.The half-life extended by herbicide mixture would affect the soil nutrients.Weed-soybean competitions affected the absorption of nutrients on soybean and yield.However,effective control of weeds could significantly reduce the weed density,improve the field through different layers of light transmittance and the absorption of nutrients on soybean,and enhance the comprehensive competitiveness of soybean.展开更多
第一代抗草甘膦转CP4-EPSPs基因大豆GTS40-3-2是国际上应用时间最长、种植面积最大的转基因作物。本文以6份GTS40-3-2衍生的抗草甘膦转基因大豆新品系为亲本,配制4个杂交组合,利用抗性分级法和相对株高法鉴定杂交亲本及其F_(2:3)子代对...第一代抗草甘膦转CP4-EPSPs基因大豆GTS40-3-2是国际上应用时间最长、种植面积最大的转基因作物。本文以6份GTS40-3-2衍生的抗草甘膦转基因大豆新品系为亲本,配制4个杂交组合,利用抗性分级法和相对株高法鉴定杂交亲本及其F_(2:3)子代对草甘膦的耐受性差异,分析其抗性水平与遗传背景的相关性。结果表明,以1230 g a.i.hm^(–2)草甘膦喷施处理时,转基因亲本及其F_(2:3)子代的苗期生长受草甘膦抑制不显著,而当喷施浓度提高至3690 g a.i.hm^(–2)和4920 g a.i.hm^(–2)时则抑制作用显著。供试的6个杂交亲本中以ZLHJ06-1568、ZLHJ10-713和ZLHJ06-698对草甘膦的耐受性相对较强,而4个F_(2:3)组合中以ZLHJ10-713×ZLHJ06-698后代在草甘膦喷施后株高受抑制最小,对草甘膦耐受性最强。不同组合后代对草甘膦的耐受性普遍优于其双亲,呈现出杂种优势。各组合后代与亲本之间对草甘膦的耐受性均呈正相关,但由于亲本间互作效应的不同,导致后代抗性水平产生差异。本研究表明草甘膦抗性基因CP4-EPSPs在大豆中的表达水平与其遗传背景相关联,为利用转基因大豆新种质培育转基因大豆新品种过程中目标基因的定向选择提供了参考依据。展开更多
为了评估转基因大豆向野大豆(Glycine soja Sieb.et Zucc.)基因漂移后可能产生的生态风险,选取抗草甘膦转基因大豆(携带外源基因cp4-epsps)与野大豆(来自内蒙古包头)杂交F_(2)代种皮颜色为黑色、褐绿色和褐黄色(与野大豆种皮颜色相近)...为了评估转基因大豆向野大豆(Glycine soja Sieb.et Zucc.)基因漂移后可能产生的生态风险,选取抗草甘膦转基因大豆(携带外源基因cp4-epsps)与野大豆(来自内蒙古包头)杂交F_(2)代种皮颜色为黑色、褐绿色和褐黄色(与野大豆种皮颜色相近)的种子,对这些种子萌发产生的F_(3)代植株的19个定量性状和5个定性性状进行分析,在此基础上,将F_(3)代植株及其亲本进行聚类分析,并对不同组F_(3)代的定量和定性性状进行分析。结果显示:F_(3)代幼苗中抗性植株与非抗性植株的分离比符合“5∶1”的孟德尔遗传定律。定量性状中,结实率的变异系数最小(4.65%),其余18个性状的变异系数高于14%,其中,单株种子数的变异系数最大(55.68%)。不同种皮颜色F_(2)代种子萌发产生的F_(3)代植株均以强缠绕型占比最高;F_(3)代植株中,90.7%的果荚为弯镰形或弓形,且80.0%的果荚颜色为黑褐色;黑色种皮F_(2)代种子萌发产生的F_(3)代种皮颜色无性状分离,褐绿色和褐黄色种皮F_(2)代种子萌发产生的F_(3)代种皮颜色均出现性状分离;F_(3)代种子中,82.7%的种子有泥膜。聚类结果显示F_(3)代植株被分为3组。不同组F_(3)代中,Ⅰ组的多数性状最低,且株高等5个性状显著低于野大豆;Ⅱ组的9个性状最高,且单株结荚数等15个性状显著高于野大豆,仅株高显著低于野大豆;Ⅲ组的部分性状最高,且单株结荚数等12个性状显著高于野大豆,株高等3个性状显著低于野大豆。综合比较认为,Ⅱ组的竞争能力最强,Ⅰ组的竞争能力最弱。Ⅰ组的不缠绕型植株占比最高,而Ⅱ和Ⅲ组的强缠绕型植株占比最高。3组F_(3)代的种皮颜色均以黑色占比最高。综上所述,供试的不同组F_(3)代植株均能完成生活史并产生后代。若抗草甘膦转基因大豆的基因通过花粉漂移到野大豆上并成功杂交,则会产生不同适应性杂交后代,增加生态风险。展开更多
文摘A PCR-ELISA method for detecting the glyphosate resistant transgenic soybean was established and optimized. The results showed that the key parameters of PCR-ELISA were as follows： the concentration of digoxin tag probe was 0.5 μmol · L^-1, the time of hybridization reaction was 15 min and the chromogenic reaction should last for 30 min. The sensitivity and the repeatability of our PCR-ELISA method were evaluated, and the results showed that it could be detected when the concentration of DNA template from transgenic soybean samples was 0.01% or higher, and the coefficient of variation of this method was less than 5% in our research condition. These results suggested that PCR-ELISA method establishment in this study had good repeatability and high precision for detecting the transgenic soybean samples.
基金Supported by Environmental Safety Assessment of Key Research Projects of Ministry of Agriculture(2016ZX08011-003)
文摘Competitions of light,nutrient and water between transgenic glyphosate resistant soybean and weed were evaluated with different herbicides(glyphosate,acetochlor,and the mixture of fomesafen and quizalofop-p-ethyl)in filed,which displayed that the application of the mixture of fomesafen and quizalofop-p-ethyl on soybeans had the best anti-herbicidal effect while glyphosate had the weakest effect.The half-life extended by herbicide mixture would affect the soil nutrients.Weed-soybean competitions affected the absorption of nutrients on soybean and yield.However,effective control of weeds could significantly reduce the weed density,improve the field through different layers of light transmittance and the absorption of nutrients on soybean,and enhance the comprehensive competitiveness of soybean.
基金国家转基因生物新品种培育重大专项(ZX2008004-001ZX2011004-001+5 种基金ZX2013004-001ZX2014004-001)资助supported by the Major Project of China on New Varieties of GMO Cultivation(ZX2008004-001ZX2011004-001ZX2013004-001ZX2014004-001)
文摘第一代抗草甘膦转CP4-EPSPs基因大豆GTS40-3-2是国际上应用时间最长、种植面积最大的转基因作物。本文以6份GTS40-3-2衍生的抗草甘膦转基因大豆新品系为亲本,配制4个杂交组合,利用抗性分级法和相对株高法鉴定杂交亲本及其F_(2:3)子代对草甘膦的耐受性差异,分析其抗性水平与遗传背景的相关性。结果表明,以1230 g a.i.hm^(–2)草甘膦喷施处理时,转基因亲本及其F_(2:3)子代的苗期生长受草甘膦抑制不显著,而当喷施浓度提高至3690 g a.i.hm^(–2)和4920 g a.i.hm^(–2)时则抑制作用显著。供试的6个杂交亲本中以ZLHJ06-1568、ZLHJ10-713和ZLHJ06-698对草甘膦的耐受性相对较强,而4个F_(2:3)组合中以ZLHJ10-713×ZLHJ06-698后代在草甘膦喷施后株高受抑制最小,对草甘膦耐受性最强。不同组合后代对草甘膦的耐受性普遍优于其双亲,呈现出杂种优势。各组合后代与亲本之间对草甘膦的耐受性均呈正相关,但由于亲本间互作效应的不同,导致后代抗性水平产生差异。本研究表明草甘膦抗性基因CP4-EPSPs在大豆中的表达水平与其遗传背景相关联,为利用转基因大豆新种质培育转基因大豆新品种过程中目标基因的定向选择提供了参考依据。
文摘为了评估转基因大豆向野大豆(Glycine soja Sieb.et Zucc.)基因漂移后可能产生的生态风险,选取抗草甘膦转基因大豆(携带外源基因cp4-epsps)与野大豆(来自内蒙古包头)杂交F_(2)代种皮颜色为黑色、褐绿色和褐黄色(与野大豆种皮颜色相近)的种子,对这些种子萌发产生的F_(3)代植株的19个定量性状和5个定性性状进行分析,在此基础上,将F_(3)代植株及其亲本进行聚类分析,并对不同组F_(3)代的定量和定性性状进行分析。结果显示:F_(3)代幼苗中抗性植株与非抗性植株的分离比符合“5∶1”的孟德尔遗传定律。定量性状中,结实率的变异系数最小(4.65%),其余18个性状的变异系数高于14%,其中,单株种子数的变异系数最大(55.68%)。不同种皮颜色F_(2)代种子萌发产生的F_(3)代植株均以强缠绕型占比最高;F_(3)代植株中,90.7%的果荚为弯镰形或弓形,且80.0%的果荚颜色为黑褐色;黑色种皮F_(2)代种子萌发产生的F_(3)代种皮颜色无性状分离,褐绿色和褐黄色种皮F_(2)代种子萌发产生的F_(3)代种皮颜色均出现性状分离;F_(3)代种子中,82.7%的种子有泥膜。聚类结果显示F_(3)代植株被分为3组。不同组F_(3)代中,Ⅰ组的多数性状最低,且株高等5个性状显著低于野大豆;Ⅱ组的9个性状最高,且单株结荚数等15个性状显著高于野大豆,仅株高显著低于野大豆;Ⅲ组的部分性状最高,且单株结荚数等12个性状显著高于野大豆,株高等3个性状显著低于野大豆。综合比较认为,Ⅱ组的竞争能力最强,Ⅰ组的竞争能力最弱。Ⅰ组的不缠绕型植株占比最高,而Ⅱ和Ⅲ组的强缠绕型植株占比最高。3组F_(3)代的种皮颜色均以黑色占比最高。综上所述,供试的不同组F_(3)代植株均能完成生活史并产生后代。若抗草甘膦转基因大豆的基因通过花粉漂移到野大豆上并成功杂交,则会产生不同适应性杂交后代,增加生态风险。
