甜瓜黄斑病毒SYBR Green Ⅰ实时荧光定量PCR方法     

孙晓辉  潘睿婧  刘永光  王丹  石朝鹏  乔宁  孙作文  竺晓平 《植物病理学报》2023,53(1):119-125

为建立检测甜瓜黄斑病毒(melon yellow spot virus, MYSV)的SYBR Green Ⅰ实时荧光定量PCR(qPCR)方法。基于MYSV核衣壳蛋白基因保守序列设计qPCR特异性引物对,针对引物退火温度、引物浓度、特异性和敏感性进行系列优化。结果显示,优化后的qPCR方法最适退火温度为61.3℃,最适引物浓度为0.65μmol·L^-1,特异性强,灵敏度高,比PCR高100倍。以携带目的基因片段的重组质粒为标准品,构建的qPCR标准曲线循环阈值与模板浓度呈良好的线性关系,相关系数为0.999 7。实验样品验证表明建立的qPCR方法可用于MYSV的定量检测。  相似文献   

薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用     

赵玉强  卓可儿  郭云  朱灿灿  王行军  陆小美  田艳丽  胡白石 《植物保护》2023,49(3):193-198

疮痂病是薄壳山核桃上最具毁灭性的病害,带菌植物材料是传播疮痂病的重要来源。准确、灵敏、快速的检测方法可为该病害流行规律调查和防控提供有力的依据。本文通过比较薄壳山核桃疮痂病菌Venturia effusa及其近似种之间的ITS序列差异,设计了特异性引物和TaqMan探针,建立了薄壳山核桃疮痂病菌的荧光定量PCR检测方法。特异性检测结果表明,该方法可以检测不同地区的薄壳山核桃疮痂病菌菌株,而对其近似种以及薄壳山核桃上的其他真菌均没有信号。本研究建立的检测方法对薄壳山核桃疮痂病菌DNA的最低检测限可达0.5 pg/μL。该方法用于田间样品检测时,检测时间仅需1 h,远快于常规的分离培养法。本研究建立的基于TaqMan探针的荧光定量PCR检测方法为薄壳山核桃疮痂病菌的快速检测和监测提供了有力工具。  相似文献   

Assessment of microbial biocontrol agent (BCA) viability to mechanical and thermal stress by simulating spray application conditions     

Marco Grella  Fabrizio Gioelli  Paolo Marucco  Ingrid Zwertvaegher  Eric Mozzanini  Marco Pittarello  Paolo Balsari  Spyros Fountas  David Nuyttens  Monica Mezzalama  Massimo Pugliese 《Pest management science》2023,79(11):4423-4438

Background

In order to improve the biological control agent (BCA) efficacy, stress factors threatening the viability of microorganisms during spray application need to be determined. The effect of spray mixture temperature and exposure time on Trichoderma harzianum T 22 and Bacillus amyloliquefaciens QST713 viability were tested. Concurrently the combined effect of mechanical and thermal stress effect on BCA viability were tested at two initial spray mixture temperatures (14 and 25 °C) by simulating a spray application using airblast sprayers featured by different tank capacity and a spray liquid circuit (without and with hydraulic agitation system). To assess the BCA microorganism viability, spray mixture samples were collected at time intervals along trials and plated to count the colony forming units (CFU).

Results

The critical temperature threshold that inhibited BCA viability was 35 °C with 30 min of exposure. The sprayer type, the initial temperature of the spray mixture and the temperature increment during the trials significantly decreased the number of CFU recovered. When simulating a spray application, the spray mixture temperature increase rate was determined mainly by the residual amount of spray mixture in the tank. Even if the tank capacity does not substantially affect the final temperature reached by the spray mixture, the higher residual spray mixture in bigger tanks can expose the BCAs for a longer time to critical temperatures.

Conclusions

Experimental trials allowed us to identify the effect of factors affecting the viability of tested BCAs, providing information about the actual chance to guarantee the biological efficacy of BCA treatments. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.  相似文献   

高端牛奶的市场竞争环境分析     

王力纬  张宇  高颖智 《中国乳业》2023,(4):2-10

在消费升级和国民健康意识的普及趋势下,我国乳制品消费量也逐渐提高,高端牛奶在特仑苏和金典广泛的市场教育下,迎来了稳定增长。牛奶高端化大势所趋,地方品牌在高端市场、超高端市场为消费者提供了更多选择。这一点线下市场尤其明显,线下市场地方品牌的高端奶占据了2/3以上的市场份额,“特金”一统高端市场的时代正在成为过去式。同时,地方品牌的活跃和高端产品的多元化也充分说明统一标准化与天然概念不能兼得。  相似文献   

鼠源细胞TaqMan实时荧光定量PCR检测方法的建立     

苏佳  赵炜  秦义娴  邓永  高月异  白洪旭  薛青红  陈晓春  吴华伟 《中国兽药杂志》2023,57(3):10-16

为对鼠源细胞进行鉴别检测,以鼠线粒体16S rRNA基因序列为靶位点设计特异性引物及探针,建立实时荧光定量PCR检测方法,并评价该方法的特异性及敏感性。结果显示,所建立的检测方法特异性好,针对鼠源细胞基因组荧光定量PCR扩增曲线良好,其他物种来源细胞基因组及生物制品原辅材料未出现特异性扩增曲线;敏感性高,基因拷贝数检出限度为45.3拷贝。本试验建立的荧光定量PCR检测方法能够有效地对鼠源细胞进行快速检测,为细胞质量控制提供了有效方法。  相似文献   

玉米转Bt基因自交系的抗玉米螟特性鉴定初报   总被引：7，自引：0，他引：7  

刘宗华  汤继华  李宝健  范云六  李桂玲  季良越  陈伟程 《作物学报》2003,29(4):621-625

通过田间接卵和室内PCR分子检测,对采用回交法育成的Mo17Bt、478Bt、综3Bt和87-1Bt等4个转Bt基因自交系的抗虫性以及Bt基因的存在,进行鉴定.结果表明,4个转基因自交系群体的抗虫性明显提高,虫孔隧道长度比阴性对照平均缩短49.70%,差异达显著水平,但抗虫性不如阳性对照;群体内,单株间抗虫性仍存在差异,有高抗到高感不同类型,其  相似文献   

植物基因启动子的克隆方法及其应用   总被引：1，自引：0，他引：1  

尹辉　李丹张毅  李秋莉 《分子植物育种》2006,4(Z1):85-91

植物基因的表达调控已成为分子生物学研究热点,启动子是基因表达调控的重要顺式元件,启动子的克隆对于研究基因表达调控、构建基因工程载体、表达目的蛋白有着重要的意义。启动子克隆的方法很多,从常用的启动子陷阱技术筛选启动子到PCR方法的应用,此后相继问世的一些基于PCR的克隆启动子技术,如载体锚定PCR、反向PCR、接头PCR、交错热不对称PCR等,为克隆启动子提供了更可靠,更合理的方法。本文着重介绍了几种植物基因启动子的克隆方法,分析了它们的优缺点,并展望了今后的研究前景。  相似文献   

基于势场蚁群算法的移动机器人全局路径规划方法   总被引：2，自引：0，他引：2  

刘建华  杨建国  刘华平  耿鹏  高蒙 《农业机械学报》2015,46(9):18-27

针对移动机器人路径规划蚁群算法收敛速度慢和人工势场法易陷入局部最优的问题,提出一种以栅格地图为环境模型,在蚁群算法搜索过程中加入针对具体问题的人工势场局部搜索寻优算法,将人工势场法中力因素转换为局部扩散信息素,使蚁群倾向于具有高适应值的子空间搜索,减少了蚁群算法在盲目搜索路径过程中产生的局部交叉路径及蚂蚁"迷失"数量,提高了蚁群对障碍物的预避障能力。对不同参数组合下2种算法及其它改进算法仿真结果做了比较,验证了基于势场蚁群算法的全局路径规划能够加快寻优过程且具有较强的搜索能力,收敛速度提高近1倍。  相似文献   

基于改进蚁群算法的3-PPR并联机构位置正解研究   总被引：1，自引：0，他引：1  

吴小勇  谢志江  宋代平  刘飞  罗久飞  毛冰滟 《农业机械学报》2015,46(7):339-344

针对3-PPR并联机构位置正解问题,运用螺旋理论计算其自由度,并分析其运动特性,同时通过解析法构建该并联机构的完整运动学逆解。将并联机构运动学正解问题的求解转化为对目标函数优化问题的求解,并基于此建立该并联机构正解的目标函数优化模型。通过对基本蚁群算法进行有效的改进,构建了可用于求解连续优化问题的改进蚁群算法。运用该算法求解3-PPR并联机构的运动学正解并进行Matlab仿真分析,对比传统数值方法和改进蚁群算法,证实该算法具有良好的全局寻优能力,并能避免初值和局部最优值对计算结果的影响,可有效应用于求解并联机构的位置正解问题。  相似文献   

Determination of vc and mating types of Cryphonectria parasitica isolates by multiplex PCR and their genetic diversity in 13 chestnut-growing provinces of Turkey     

Deniz Çakar  Göksel Özer  Seçil Akıllı Şimşek  Salih Maden 《Forest Pathology》2023,53(3):e12813

Vegetative compatibility (vc) and mating types and genetic diversity of Cryphonectria parasitica isolates were determined using 183 isolates obtained from 215 infected chestnut trees growing in 13 provinces of Turkey. Based on the cultural aspects, 143 of these isolates were evaluated as virulent whereas the remaining 40 isolates were hypovirulent. When vc types of 183 isolates were classically differentiated, 135 of them matched to EU-1 (82.3%), 29 of them to EU-12 (17.6%) vc type, whereas 19 of them did not match to the two. When molecular vic markers were used, all the isolates were assigned to two EU vc types; 149 to EU-1 (81.4%) and 34 (18.5%) to EU-12. Of the majority of the isolates, 134 (73.2%) had mating-type MAT-1, while 44 (24%) isolates had MAT-2 and 5 (2.8%) isolates had both mating types. The population analysis based on two DNA marker systems, Inter-Primer Binding Site and Start Codon Targeted Polymorphism, showed no intraspecific genetic variation among the C. parasitica isolates. The prevalence of two dominant vc types revealed by this study shows that biological control with hypovirulent EU-1 and EU-12 isolates will be significant for the country. The results might be helpful to chestnut breeders carrying out resistance breeding studies to manage this disease based on hypovirulence attributed to Cryphonectria hypovirus 1.  相似文献