Event-specific quantitative detection of nine genetically modified maizes using one novel standard reference molecule   总被引：2，自引：0，他引：2  

Yang L  Guo J  Pan A  Zhang H  Zhang K  Wang Z  Zhang D 《Journal of agricultural and food chemistry》2007,55(1):15-24

With the development of genetically modified organism (GMO) detection techniques, the Polymerase Chain Reaction (PCR) technique has been the mainstay for GMO detection, and real-time PCR is the most effective and important method for GMO quantification. An event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity. This study establishes the event-specific detection methods for TC1507 and CBH351 maizes. In addition, the event-specific TaqMan real-time PCR detection methods for another seven GM maize events (Bt11, Bt176, GA21, MON810, MON863, NK603, and T25) were systematically optimized and developed. In these PCR assays, the fluorescent quencher, TAMRA, was dyed on the T-base of the probe at the internal position to improve the intensity of the fluorescent signal. To overcome the difficulties in obtaining the certified reference materials of these GM maizes, one novel standard reference molecule containing all nine specific integration junction sequences of these GM maizes and the maize endogenous reference gene, zSSIIb, was constructed and used for quantitative analysis. The limits of detection of these methods were 20 copies for these different GM maizes, the limits of quantitation were about 20 copies, and the dynamic ranges for quantification were from 0.05 to 100% in 100 ng of DNA template. Furthermore, nine groups of the mixed maize samples of these nine GM maize events were quantitatively analyzed to evaluate the accuracy and precision. The accuracy expressed as bias varied from 0.67 to 28.00% for the nine tested groups of GM maize samples, and the precision expressed as relative standard deviations was from 0.83 to 26.20%. All of these indicated that the established event-specific real-time PCR detection systems and the reference molecule in this study are suitable for the identification and quantification of these GM maizes.  相似文献   

Improvement of texture properties and flavor of frozen dough by carrot (Daucus carota) antifreeze protein supplementation     

Zhang C  Zhang H  Wang L  Gao H  Guo XN  Yao HY 《Journal of agricultural and food chemistry》2007,55(23):9620-9626

The effects of concentrated carrot protein (CCP) containing 15.4% (w/w) carrot (Daucus carota) antifreeze protein on texture properties of frozen dough and volatile compounds of crumb were studied. CCP supplementation lowered the freezable water content of the dough, resulting in some beneficial effects including holding loaf volume steadily and making the dough softer and steadier during frozen storage. Furthermore, SPME-GC-MS analysis showed CCP supplementation did not give any negative influences on volatile compounds of crumb and gave a pleasant aroma felt like Michelia alba DC from trans-caryophyllene simultaneously. Combining our previous results that CCP supplementation improves the fermentation capacity of the frozen dough, CCP could be used as a beneficial additive for frozen dough processing.  相似文献   

Flumorph Is a Novel Fungicide That Disrupts Microfilament Organization in Phytophthora melonis     

Sheng Zhu S  Li Liu X  Fei Liu P  Li Y  Qiang Li J  Min Wang H  Kui Yuan S  Guo Si N 《Phytopathology》2007,97(5):643-649

ABSTRACT The mechanism of the effects of flumorph (a novel fungicide) was investigated by analyzing alterations of hyphal morphology, cell wall deposition patterns, F-actin organization, and other organelles in Phytophthora melonis. Calcofluor white staining suggested that flumorph did not inhibit the synthesis of cell wall materials, but disturbed the polar deposition of newly synthesized cell wall materials during cystospore germination and hyphal growth. After exposure to flumorph, zoospores were able to switch into cystospores accompanied with the formation of a cell wall, whereas cystospores failed to induce the isotropic-polar switch and did not produce germ tubes but continued the isotropic growth phase. In flumorph-treated hyphae, the most characteristic change was the development of periodic swelling ("beaded" morphology) and the disruption of tip growth. Newly synthesized cell wall materials were deposited uniformly throughout the diffuse expanded region of hyphae, in contrast to their normal polarized patterns of deposition. These alterations were the result of F-actin disruption, identified with the fluorescein isothiocynate (FITC)-phalloidin staining. The disruption of F-actin also was accompanied by disorganized organelles: each swelling of subapical hyphae was associated with a nucleus. Vesicles did not undergo polarized secretion to the apical hyphae, but diffused around nuclei for the subapical growth; thus, the cell wall was thickened with periodic expansion along the hyphae. Upon removing flumorph, normal tip growth and organized F-actin were observed again. These data, as well as data published earlier, suggest that flumorph may be involved in the impairment of cell polar growth through directly or indirectly disrupting the organization of F-actin. The primary site of action by flumorph in the disruption of the F-actin organization is under investigation.  相似文献   

7种睡莲属植物叶绿体基因组密码子偏好性分析     

毛立彦  黄秋伟  龙凌云  檀小辉  谢红辉  唐毓玮  丁丽琼 《西北林学院学报》2022,37(2):98-107

以7种睡莲属植物叶绿体基因组为研究对象,分析其叶绿体基因组的密码子使用偏好性,探讨影响其密码子偏好性形成的主要因素。结果表明,7种睡莲叶绿体密码子使用偏好性相似,基因组序列富含A/T碱基,密码子第3位碱基以A/T结尾为主;有效密码子数（ENc）分别为40.14~61.00、40.14~61.00、40.55~57.61、39.77~57.61、39.55~57.30、39.56~61.00和40.10~61.00,均明显大于35,表明其密码子使用偏好性较弱。ENc-plot、PR2-plot和中性绘图分析表明,它们的叶绿体密码子偏好性形成受自然选择、碱基突变等多种因素共同影响,其中自然选择起主导作用。通过同义密码子RSCU值分析,发现以A/T结尾的高频密码子27个,以G/C结尾的低频密码子26个,进一步依据密码子RSCU和△RSCU值,最终确定出4个最优密码子。此外,基于7种睡莲叶绿体基因组密码子使用偏好特征RSCU值、CDS序列及其全基因组序列分别构建系统进化树,分析发现3种方法构建的物种间进化关系相似,均划分为2大分支,其中Nymphaea alba var.rubra、Nymphaea alba、Nymphaea ampla、Nymphaea capensis、Nymphaea jamesoniana和Nymphaea lotus划归第1分支,Nymphaea mexicana单独形成第2分支。研究结果进一步支持了物种内遗传关系与密码子使用偏好性之间存在一定关系。  相似文献   

Rapid evolution of Ophraella communa cold tolerance in new low-temperature environments     

Tian  Zhenqi  Chen  Guangmei  Zhang  Yan  Ma  Chao  Tian  Zhenya  Gao  Xuyuan  Chen  Hongsong  Guo  Jianying  Zhou  Zhongshi 《Journal of pest science》2022,95(3):1233-1244

Low winter temperatures severely stress newly arriving insect species. Adaptive evolutionary changes in cold tolerance can facilitate their establishment in new environments. Ambrosia artemisiifolia, a noxious invasive plant, occurs throughout China. Ophraella communa, a biological control agent of A. artemisiifolia, mainly occurs in southern China. However, in 2012, it established populations in Beijing (39.98°N, 115.97°E) following introduction from Laibin (23.62°N, 109.37°E), implying cold adaptation. The mechanisms underlying its rapid evolution of cold tolerance remain unknown. We investigated the levels of cryoprotectants and energy reserves in adult O. communa from two latitudes. In high-latitude insects, we found high trehalose, proline, glycerol, total sugar, and lipid levels; five potential genes (Tret1a, Tret1b, Tret1-2, P5CS, and GST), responsible for regulating cold tolerance and involved in trehalose transport, proline biosynthesis, and glutathione S-transferase activation, were highly expressed. These hybridisation changes could facilitate cold temperature adaptation. We demonstrate the genetic basis underlying rapid adaptation of cold tolerance in O. communa, explaining its extension to higher latitudes. Thus, specialist herbivores can follow host plants by adapting to new temperature environments via rapid genetic evolution.

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武汉市农田林网白蚁生存环境与防治技术研究初探     

王菊香谢新国郭红艳  郑淑芳胡进 洪承昊 《湖北林业科技》2022,(1):28-31

白蚁是危害武汉市农田林网的主要害虫之一,对全市立木、农作物、建筑和木材谷物等造成很大损失。本研究调查了武汉市主要农田林网地区白蚁的种类、生物学特性及活动生存环境,并分析了种群与土壤之间的关系。结果显示项目研究地农田林网树木白蚁主要为黑翅土白蚁,喜欢生活在潮湿的土壤中。在粘壤土中,主巢入土深度在150 cm以内的占总数80.2%。通过毒饵诱杀法和药物阻隔法,市售的三种药剂均对黑翅土白蚁具有较好的防治效果,平均防治效果达到80.0%以上。  相似文献   

Long-term field evaluation and large-scale application of a Metarhizium anisopliae strain for controlling major rice pests     

Peng  Guoxiong  Xie  Jiaqin  Guo  Rong  Keyhani  Nemat O.  Zeng  Deyu  Yang  Puyun  Xia  Yuxian 《Journal of pest science》2021,94(3):969-980

Journal of Pest Science - There is increasing concern about the toxic effects of chemical pesticides on human health and the environment. Many alternatives, however, are viewed as impracticable or...  相似文献   

重庆地区叶用枸杞“宁杞9号”扦插育苗技术研究     

李果  李承承  湛小燕  王云堂 《绿色科技》2021,(3)

为了探究适宜重庆地区的叶用枸杞“宁杞9号”扦插育苗技术,进行了不同扦插基地配比处理下的叶用枸杞扦插育苗技术研究。结果表明:河沙∶椰棕土∶草炭土∶有机肥(3∶3∶3∶1)的基质配比扦插效果最好,“宁杞9号”扦插成活率达到90.25%,扦插苗生根数量4.05根,扦插苗新生根长4.26 cm,扦插苗株高28.39 cm,扦插苗新生枝条粗度0.43,而新生枝条数量无显著差异。  相似文献   

14-3-3蛋程中的互作靶蛋白鉴定及其对外源激素的响应     

张志兴  敏秀梅  宋果  陈花  许海龙  林文雄 《中国农业科学》2021,54(12):2523-2537

【目的】籽粒灌浆对水稻产量及品质的形成至关重要。14-3-3蛋白是一种信号转导调节因子,在植物生长发育中发挥着重要调控作用。本研究通过分析14-3-3蛋白家族在籽粒灌浆过程中的基因表达模式及其互作靶蛋白,从而揭示其在籽粒灌浆过程中的功能。【方法】利用实时荧光定量PCR（qRT-PCR）技术分析水稻14-3-3基因家族在籽粒灌浆过程中的表达变化模式,并从中选取GF14b及GF14e进行后续的蛋白功能分析。利用KEGG数据库对GF14b及GF14e蛋白功能motif位点进行分析;构建GST-GF14b及GST-GF14e表达载体,利用亲和层析技术分别钓取籽粒中与GF14b及GF14e互作的靶蛋白,并借助LC-MS/MS对靶蛋白进行鉴定。采用GST pull-down方法验证靶蛋白与GF14b及GF14e间的蛋白互作关系。利用Kinasephos在线程序对靶蛋白的Ser和Thr磷酸化位点进行预测;采用MapMan 3.6.0软件对靶蛋白的功能及参与的代谢过程进行分析。在籽粒灌浆期（花后15 d）,分别喷施25×10^-6mol·L^-1 ABA,10×10^-6mol·L^-1 IAA,100×10^-6mol·L^-1 GA,50×10^-6mol·L^-1 ZR和 2×10^-4mol·L^-1 BR,研究外源激素处理对籽粒灌浆过程中GF14b,GF14e及其互作靶基因表达的影响。【结果】14-3-3家族基因中,除GF14h外,其余7个家族成员在水稻籽粒中均有表达,其中GF14b及GF14e在籽粒灌浆过程中的表达水平较高且变化幅度较大。通过蛋白序列分析发现,GF14b与GF14e间具有3个相同,2个差异的motif功能位点。通过亲和层析试验,在籽粒中共鉴定到59个与GF14b和72个与GF14e互作的靶蛋白,其中有43个靶蛋白与2个成员均有互作,分别有16个和29个靶蛋白与GF14b和GF14e特异结合。随机选取2个靶蛋白进行体外蛋白互作验证,结果表明靶蛋白SUS3与GF14b和GF14e均存在互作关系,而靶蛋白PSA仅与GF14e有相互作用关系,验证了亲和层析结果的准确性。蛋白功能的分析表明,GF14b和GF14e通过与靶蛋白的结合,共同参与了籽粒灌浆过程中蔗糖转化、淀粉合成、糖酵解、TCA循环等碳代谢途径。同时,GF14b及GF14e还具有特异的调控功能,其中GF14b与核酸代谢及物质转运密切相关,而GF14e与C1代谢中的关键蛋白存在互作。此外,大部分靶蛋白均鉴定到具有潜在的Ser和Thr磷酸化位点。外源激素处理下,籽粒中GF14b和GF14e上调表达,而与淀粉合成代谢相关的靶基因（SUS2、 AGPS、AGPL、PPDK2、SBE）大部分呈下调表达的趋势。【结论】14-3-3基因家族成员GF14b和GF14e在水稻籽粒灌浆过程中的表达变化幅度较大,且会响应激素浓度的改变,并通过蛋白互作的形式负调控淀粉合成代谢相关基因的表达,从而对水稻籽粒淀粉的合成起到重要的调控作用。  相似文献   

不同提取方法对红松籽油提取效果及功能性质的影响     

潘晓丽  王凤娟  张娜  郭庆启 《北京林业大学学报》2021,43(1):127-135

  目的  立足于溶剂法,通过对比超声波、微波、光波单独处理较短时间的红松籽油的得率,以及处理后单独浸提红松籽油得率的增加情况,明确在红松籽油提取过程中处于主导作用的方法,同时确认4种提取方法对红松籽油的理化指标、脂肪酸和抗氧化能力的影响。  方法  按照GB/T 5009.229—2016《食品中酸价的测定》、GB/T 5532—2008《动植物油脂 碘值的测定》和GB/T 5009.227—2016《食品中过氧化值的测定》分别测定4种方法提取的红松籽油的酸值、碘值和过氧化物值,采用气相色谱–质谱联用法测定红松籽油中脂肪酸的种类和质量分数,采用分光光度法测定红松籽油对DPPH·和ABTS+·的清除能力。  结果  正己烷作为最佳提取溶剂,当提取温度25 ℃,液料比18 mL/g,浸提时间5 h红松籽油得率为65.52%。超声波法（300 W,10 min）、微波法（380 W,100 s）、光波法（400 W,11 min）单独处理时的红松籽油得率分别为67.63%、62.26%、58.25%。上述3种方法处理后再单独使用正己烷浸提,使总提取时间达到5 h后的红松籽油得率分别增加了2.18%、4.50%、6.10%。4种方法提取的红松籽油符合GB/T 2716—2018《食品安全国家标准 植物油》中酸值（≤ 4 mg/g）、过氧化值（≤ 0.25 g/100 g）的限量标准。溶剂法制备出的红松籽油的酸值和过氧化值均最低（P < 0.05）。脂肪酸质量分数由高到低分别是亚油酸（44.54% ~ 46.32%）、油酸（28.29 % ~ 28.83%）和皮诺敛酸（13.15% ~ 14.51%）。溶剂法、超声波法、微波法、光波法所提取的红松籽油清除DPPH·和ABTS+·的IC₅₀值分别是9.41、8.80、9.43、9.61 g/L和5.10、5.51、6.10、5.43 g/L。  结论  对超声波法、微波法和光波法单独作用及浸提后得率进行比较,发现超声波法、微波法和光波法在提取过程中起主导作用。与溶剂法、微波法和光波法相比,超声波法提取的红松籽油得率高,不饱和脂肪酸质量分数高,抗氧化能力强,能够达到较好的提取效果,此方法应用于红松籽制油工业中具有一定优势。   相似文献