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多重PCR检测抗草甘膦大豆GTS40-3-2 总被引：1，自引：0，他引：1
将大豆内源参照基因(Lectin)、外源基因P-CaMV 35S、CP4-EPSPS和T-NOS作为多重PCR检测参数,建立了一种多重PCR检测抗草甘膦大豆GTS40-3-2的方法.反应条件优化结果表明:多重PCR的适宜退火温度为58℃,适宜引物终浓度(μmol· L-)配比为:0.1∶0.2∶0.2∶0.2.采用已知样品对多重PCR体系进行验证,检测结果可靠,证明该多重PCR体系为一种有效的抗草甘膦大豆GTS40-3-2的检测方法. 相似文献
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对引进的美国大豆品种进行转基因成分的检测 总被引：7，自引：1，他引：6
利用田间表型鉴定和PCR检测相结合的方法,对从美国引进的46个大豆品种进行了转基因成分的检测.实验过程中,首先通过喷洒草甘膦对待测材料进行表型鉴定,然后通过PCR分子检测分析CaMV35S启动子和NOS终止子的有无,初步判定引进材料是否为转基因大豆,最后对目的基因CP4-EPSPS进行检测,进一步确定所测材料是否为抗草甘膦转基因大豆.大豆本身的凝集素基因作为PCR检测的内置检测标准,有效排除了由于DNA质量、实验操作等因素对检测结果的影响.结果表明,在所检测的46个引进品种中,5个是抗草甘膦转基因品种,41个是非转基因品种,它们可作为我国大豆新品种选育的亲本材料. 相似文献
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抗草甘膦转基因大豆及加工品LAMP检测研究 总被引：9，自引：0，他引：9
将环介导的等温核酸扩增技术应用于转基因大豆及加工品检测.针对抗草甘膦Roundup Ready转基因大豆及加工品外源基因EPSPS设计2对特异性引物进行扩增,成功建立起定性检测转基因大豆及加工品的LAMP检测方法.优化LAMP反应条件,反应温度为65℃,反应时间为1h.结果表明:该体系能快速、灵敏、有效地检测转基因大豆及加工品中整合的EPSPS基因,检测限为0.01%,低于国际现行最低检测量0.5%的要求.检测EPSPS基因操作简单,成本低、特异性强、灵敏度高.LAMP检测结果可信,稳定性好,可对目前批准的抗草甘膦RoundupReady转基因大豆及加工品进行定性检测. 相似文献
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野生大豆是大豆遗传改良的重要资源.转基因大豆可能对野生大豆资源存在潜在的农业和生态风险.外源基因从抗草甘膦转基因大豆向野生大豆材料的逃逸不仅需要成功的杂交,还要依赖于杂交后代的适合度.因此野生大豆和抗草甘膦转基因大豆杂交后代的适合度分析,对评价抗草甘膦转基因逃逸引起的生态风险非常必要.在网室条件下,4个野生大豆材料和抗草甘膦转基因大豆RR能够杂交结实,获得有抗草甘膦基因杂交后代群体F1和F2(江浦野生豆-5×RR).对杂交后代及其母本野生大豆材料的7个农艺性状进行调查,计算适合度并进行t测验分析.结果表明:在没有草甘膦的选择压力下,杂交后代在一些性状上的相对适合度高于母本野生大豆材料;江浦野生豆-5和RR杂交F2代敏感株与抗性株在7个农艺性状有相对适合度上均差异不显著. 相似文献
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改良Chelex-100法和CTAB法用于转基因抗草甘膦大豆检测效果的比较 总被引：2，自引：0，他引：2
以转基因抗草甘膦大豆为研究材料,分别采用改良Chelex-100法和常规CTAB法提取基因组DNA,以提取DNA的浓度和纯度,同时以PCR扩增大豆的内源基因(lectin)及外源特异性序列(CaMV35S,nos,Cp4-epsps)的效果对两种方法进行比较和评价.结果表明:虽然改良Chelex-100法DNA提取纯度不高,但是提取效率与常规CTAB法相当,而且改良Chelex-100法能够快速在1 h之内从大豆中提取DNA,所提取的DNA可以直接用于PCR扩增反应,PCR扩增产物电泳条带清晰.因此,改良Chelex-100法可以替代CTAB法提取DNA用于转基因检测,该方法具有经济、简便、快速的特点. 相似文献
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利用“微创刷”法获得抗草甘膦转基因大豆 总被引：2，自引：0，他引：2
以发芽3 d的大豆成熟种子胚尖生长点为作用点,利用"微创刷"法将抗草甘膦基因(EPSPS)转入绥农22中,对转化植株T1代进行草甘膦筛选,对筛选后的抗性植株进行PCR检测,得到抗草甘膦转基因大豆。同时研究了不同浓度草甘膦对野生型绥农22与抗草甘膦转基因绥农22大豆植株的影响。结果表明:绥农22 T0代成株率为97.38%,对T1代具有草甘膦抗性的植株进行PCR检测,初步证明EPSPS基因成功转入大豆中,T1代转化效率为6.20%;对野生型绥农22与"微创刷"法获得的转基因绥农22大豆在不同浓度草甘膦进行相关生理指标测定,抗草甘膦转基因绥农22大豆在不同浓度草甘膦作用下叶片叶绿素含量指数、光合速率高于野生型绥农22大豆,莽草酸含量低于野生型绥农22大豆,进一步证明了大豆抗性植株对草甘膦的抗性。 相似文献
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《大豆科学》2015,(2)
野生大豆是大豆遗传改良的重要资源。转基因大豆可能对野生大豆资源存在潜在的农业和生态风险。外源基因从抗草甘膦转基因大豆向野生大豆材料的逃逸不仅需要成功的杂交,还要依赖于杂交后代的适合度。因此野生大豆和抗草甘膦转基因大豆杂交后代的适合度分析,对评价抗草甘膦转基因逃逸引起的生态风险非常必要。在网室条件下,4个野生大豆材料和抗草甘膦转基因大豆RR能够杂交结实,获得有抗草甘膦基因杂交后代群体F1和F2(江浦野生豆-5×RR)。对杂交后代及其母本野生大豆材料的7个农艺性状进行调查,计算适合度并进行t测验分析。结果表明:在没有草甘膦的选择压力下,杂交后代在一些性状上的相对适合度高于母本野生大豆材料;江浦野生豆 －5 和 RR 杂交 F2代敏感株与抗性株在 7 个农艺性状的相对适合度上均差异不显著。 相似文献
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转基因抗草甘膦除草剂大豆GTS 40-3-2作为目前在最多国家和地区批准种植的转基因大豆,除榨取大豆油外,剩余的豆粕等副产品被用作动物饲料。通过对山西省太谷周边11个动物养殖户的饲料进行转基因标识情况专项调查,采用定性和定量PCR技术对转基因抗草甘膦除草剂大豆GTS 40-3-2含有的Ca MV35S启动子、NOS终止子、Cp4-EPSPS及大豆内标Lectin基因进行检测。结果表明:11份饲料全部含有转基因抗草甘膦除草剂大豆GTS 40-3-2成分,且其含量各不相同,其中蛋鸡饲料GTS 40-3-2转化体的含量最低,但所有饲料均无转基因标识。试验为我国转基因饲料饲用安全性评价体系提供了试验数据支持,同时也对来源于转基因作物的原材料产品的合理应用以及通过转基因饲料喂养动物所产生的农畜产品的安全性评价具有重要的现实和理论意义。 相似文献
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PCR-ELISA法对大豆品种的转基因定性检测研究 总被引：4，自引：0，他引：4
以共价交联在PCR管壁上的寡核苷酸作为固相引物进行PCR扩增,对PCR扩增的固相和液相产物分别进行杂交和凝胶电泳检测的PCR-ELISA法,对黑龙江省常规选育品种合丰35和黑农37、外源DNA直接导入大豆的分子育种所获品种黑生101、及大连进口的美国2号等大豆,进行转基因定性检测.结果表明:该方法高效可靠,可作为一种快速定性检测转基因产品的方法;检测结果:美国2号为转基因大豆,黑生101、合丰35和黑农37为非转基因大豆.上述结果对分子育种、生态保护、安全监测及对建立黑龙江省非转基因大豆生产保护区等具有重要意义. 相似文献
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耐草甘膦和耐草铵膦是转基因作物育种重要的目标性状。将耐草甘膦基因MC1-EPSPS构建到含有bar基因的植物表达载体pTF101.1中，通过农杆菌介导法转入玉米材料Hi-II中，从而获得兼具耐受草甘膦和草铵膦性状的转基因玉米材料 CM8401。目的基因PCR检测显示，MC1-EPSPS和bar基因稳定整合到玉米基因组中。目的蛋白试纸条检测结果显示，MC1-EPSPS蛋白和PAT蛋白在转基因玉米世代间中表达稳定。田间除草剂耐受性鉴定试验表明，转基因玉米CM8401对草甘膦和草铵膦都具有良好耐受性，可耐受4倍推荐中剂量的草甘膦和草铵膦。 相似文献
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A field study was conducted in 2001 and 2002 at Blackville, SC, to compare the effectiveness of soil-applied herbicide programmes followed by glyphosate with sequential glyphosate applications in 19- and 97-cm row soybean. Treatments included pre-emergence applications of flumetsulam, chlorimuron plus sulfentrazone, metribuzin, or flumioxazin all applied with S-metolachlor with a subsequent application of glyphosate applied 5 wk after soybean planting (WAP). Additional treatments included sequential applications of glyphosate at 3 and 5 WAP and a non-treated control. Weed control was comparable in wide and narrow rows, and soil-applied herbicides were similar to sequential glyphosate applications in either row width. Amaranthus palmeri S. Wats., Digitaria sanguinalis (L.) Scop., and Cyperus rotundus L. control with all herbicides was 98–100%. All herbicide programmes provided at least 92% Ipomoea hederacea var. integriuscula Gray and I. lacunosa L. control. S-metolachlor plus flumetsulam followed by glyphosate provided 99% Senna obtusifolia (L.) Irwin and Barneby control, which was superior to the 88% and 90% obtained with chlorimuron plus sulfentrazone and metribuzin programmes, respectively. Flumioxazin and sulfentrazone caused up to 12% soybean injury. In 2001, soybean yields and gross profit margins were similar for soil-applied programmes followed by glyphosate and sequential glyphosate applications in wide and narrow rows. Conversely, soybean yields and profit margins in 2002 averaged across soil-applied programs were 380 kg/ha and $50/ha superior to sequential glyphosate applications in wide rows, but were similar in narrow rows. This research indicates a highly efficacious soil-applied herbicide program followed by glyphosate provides similar weed control to sequential glyphosate applications in wide- and narrow-row soybean; however, a soil-applied herbicide programme followed by glyphosate may improve yields and returns of wide-row soybean over sequential glyphosate applications alone. 相似文献
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A field study was conducted in 2001 and 2002 at Blackville, SC, to evaluate broadleaved weed control and economics of conventional and glyphosate-containing herbicide programmes in glyphosate-resistant soybean planted in 97-cm-wide rows. Treatments included chlorimuron plus sulfentrazone or chlorimuron plus metribuzin applied pre-emergence followed by post-emergence applications of lactofen or glyphosate 4 wk after soybean emergence (WAE). Other treatments were glyphosate applied 4 WAE, sequential applications of low dose glyphosate applied 2 and 4 WAE, and a non-treated control. Pre-emergence herbicides followed by glyphosate, controlled Ipomoea lacunosa L. 8 WAE. I. hederacea var. integriuscula Gray control with pre-emergence herbicides followed by glyphosate was 100% with similar control from chlorimuron plus sulfentrazone followed by lactofen, whereas control following the single glyphosate application was 84%. Amaranthus palmeri S. Wats. control 9 WAE was 100% following single or sequential glyphosate applications, while control ranged from 76% to 96% with pre-emergence herbicides followed by lactofen. However, early season weed interference when a single application of glyphosate was delayed until 4 WAE reduced soybean yields an average of 389 kg/ha compared to pre-emergence herbicides followed by glyphosate. Based on mean contrasts, pre-emergence herbicides followed by glyphosate had greater gross profit margins than a single glyphosate application alone, but were similar to sequential glyphosate applications. Gross profit margins from pre-emergence herbicides followed by lactofen were not different from glyphosate alone. 相似文献
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From their introduction in 1996, glyphosate resistant (GR) soybean cultivars have been rapidly adopted by farmers in Argentina and in other countries in the world. The high rate of adoption of this technology seems to be based on the simplicity of use provided by a single herbicide (glyphosate), its high efficacy to control many weeds and, the low costs of the technology relative to that used in conventional crops. During 2001–2002, 2002–2003 and 2003–2004 soybean growing seasons, field surveys and field experiments were performed with the aim of studying the effect of different glyphosate management strategies on the diversity of soybean weed communities, weed control, individual survival, fecundity and crop yield. In addition, the emergence pattern of three important weeds, Digitaria sanguinalis, Cyperus rotundus and Anoda cristata, was also studied. Both field surveys and field experiments were carried out on no-till soybean crops sown immediately after wheat or barley harvest (double cropped system). Experiments were set up in commercial soybean crops and consisted of different times of a single glyphosate application, two glyphosate applications and also the application of glyphosate plus a residual herbicide imazethapyr. A. cristata, D. sanguinalis, Stellaria media, Chenopodium album and Cyperus sp. were the most prevalent weeds recorded at pre-harvest of the soybean crops, showing regional constancy higher than 80% in both years. In three out of four field experiments, crop yield was not increased when glyphosate was applied twice compared with a single application of the herbicide. In addition there was a lower negative effect on weed species richness when glyphosate was applied once during the crop cycle than with two applications of glyphosate or glyphosate plus imazethapyr. D. sanguinalis escaped the glyphosate early treatment because of the long weed emergence period, while A. cristata and C. rotundus survived treatments due to their high individual tolerance. The results suggest that it is possible to manage glyphosate application to get high crop yield with a low impact on weed diversity, depending on the weed species and their abundance. 相似文献
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转基因抗草甘膦油菜籽中草甘膦氧化还原酶基因的检测方法研究 总被引：15，自引：1，他引：15
应用PCR技术，分析草某膦氧化还原酶基因（GOX）和修饰草甘膦氧化还原酶基因的核苷酸序列及表达蛋白质的氨基酸序列，建立了检测转基因抗草甘膦油菜籽中草甘膦氧化还原酶基因的技术和方法。 相似文献
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为方便HAL1转基因大豆的研究和检测,以转HAL1基因大豆T3代基因组DNA为模板,使用热不对称交错PCR(TAIL-PCR)方法分离其外源基因插入位点的侧翼序列,获得了插入片段DNA序列(T-DNA)的左翼序列和右翼序列。通过比对大豆基因组序列确定其整合位点,确定了HAL1基因的T-DNA在大豆基因组1号染色体非编码区49468395位点以单拷贝插入,转基因事件不影响大豆基因组功能基因的正常表达,而且在200 mmol·L~(-1)NaCl盐胁迫下转基因植株生长力明显强于对照材料。蛋白定量检测结果表明,在叶片中蛋白含量最高可达0.03 mg·g~(-1),在根茎花中也有表达,但是含量较低。根据整合位点处的左右侧翼序列设计两对特异性PCR检测引物,PCR检测结果显示只有转HAL1基因大豆阳性材料才能扩增出926和816 bp DNA片段。本研究建立的转HAL1基因大豆特异性定性检测方法,对转基因大豆的研究具有重要的意义,也为大豆转基因受体材料的管理与检测提供参考。 相似文献
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The effect of exclusive application of glyphosate and glyphosate in combination with residual herbicides on weed species density and composition in summer crops was studied over 2 years. Field experiments consisted of three rotations including soybean and maize glyphosate-resistant cultivars in two tillage systems. Regardless of the tillage system, both glyphosate application alone and in combination with residual herbicides had a pronounced impact on the reduction in density and richness of summer herbaceous annual weeds, but the effect was greater with glyphosate in combination with residual herbicides. 相似文献