共查询到18条相似文献，搜索用时 153 毫秒
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基于PCR技术的植物病原真菌检测技术研究进展 总被引：4，自引：0，他引：4
植物病原真菌是可以在植物上引起病害的一类重要病原物,对该类病原的快速检测是对其进行植物检疫、监测预报及病害防治必不可少的基础工作.近年来,以PCR为基础的分子检测技术的日益发展使得植物病原真菌的检测更加快速、灵敏和可靠.本文对近年来基于PCR技术的植物病原真菌检测方法进行了介绍与评述,这些方法包括ITS-PCR、巢式PCR、多重PCR、PCR ELISA和实时荧光定量PCR技术. 相似文献
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基于PCR技术的植物病原菌分子定量检测技术研究进展 总被引：2，自引：0，他引：2
植物病原菌的菌源量是病害发生和流行的重要因子之一,对其精准的定量测定或检测可大大提高植物病害预测的准确性,本文对实时荧光定量PCR (qPCR)与数字PCR在植物病原菌定量检测、以及基于RNA水平的real-time PCR和基于核酸染料(EMA/PMA)与qPCR相结合的技术在植物病原菌活体定量检测中的应用进行了综述,并展望其在植物病害流行和预测中的应用前景。 相似文献
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利用实时荧光定量PCR 技术检测油菜菌核病菌 总被引：2，自引：0，他引：2
采用高通量实时荧光定量PCR 技术建立检测和监测油菜菌核病菌群体数量的方法。利用油菜菌核病菌(Sclerotiniasclerotiorum)的茁-微管蛋白基因内含子序列的特异性,设计引物对SclSF (5'-CTCAAATCTCCGAAAGTT -3') / SclAF (5'-TGCAGACGGGTAATATG -3'),建立和优化了SYBR Green 玉实时荧光定量PCR 检测体系。结果表明,该引物对能够从8种所测试的十字花科植物常见病原真菌中特异性扩增出油菜菌核病菌;所建立的实时定量PCR 技术可应用于油菜病叶和病茎中菌核病菌的早期检测及菌核病的预测预报。 相似文献
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甘蔗宿根矮化病菌实时荧光定量PCR检测方法的建立 总被引：1，自引：0，他引：1
甘蔗宿根矮化病是由Leifsonia xyli subsp. xyli (Lxx)引起的一种世界性甘蔗细菌病害。根据Lxx的Pat1基因保守序列,设计并合成了一对特异性引物Pat1F (5′-GGTTCCATTGCTTACCGATT-3′)/Pat1R(5′-CAAGTTTCGACAGGAACAGC-3′),和一条TaqMan探针(FAM-5′-CCACGGCTACGTCAATTCGGG-3′-TAMRA),建立了一种特异性强、灵敏度高的甘蔗宿根矮化病菌实时荧光定量PCR检测方法。结果表明,本研究建立的实时荧光定量PCR方法,对Lxx的检测最低下限为102 copies·μL-1。应用实时荧光定量PCR与常规PCR方法对14个甘蔗品种进行Lxx检测,阳性检出率分别为86%和43%,表明实时荧光定量PCR比常规PCR检测方法具有更高的灵敏度。研究结果为甘蔗宿根矮化病的诊断、田间发生动态监测、脱毒健康种苗检测及品种/材料交换检疫检测提供了新技术支撑。 相似文献
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三种PCR方法检测柑橘黄龙病菌的效果比较 总被引：1，自引：0，他引：1
为了比较常规PCR、巢式PCR和实时荧光定量PCR方法在大田检测中对柑橘黄龙病(Huanglongbing, HLB)的检测效果, 首先比较了3种检测方法对柑橘黄龙病菌检测的灵敏度, 结果发现：3种检测方法的灵敏度依次为常规PCR<巢式PCR<实时荧光定量PCR。运用3种检测方法对广东5个柑橘品种上的189个黄龙病疑似病样进行检测, 结果发现：黄龙病检出率依次为常规PCR<巢式PCR<实时荧光定量PCR。研究表明：常规PCR适合以较低成本大规模检测黄龙病; 实时荧光定量PCR具有最大的检测灵敏度; 巢式PCR检测技术同时具有前两者的一些优点, 但操作较复杂, 适合技术熟练的研究者使用。 相似文献
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几种常用植物病原细菌分子检测方法 总被引：3，自引：0，他引：3
植物病原细菌(phytobacteria)是植物上一类重要的病原菌,这些细菌能引起多种农作物、经济作物、花卉、树木及牧草上的病害。它的快速检测是病害防治、预测预报及植物检疫必不可少的重要工作。其中,以PCR为基础的分子检测技术的进步使植物病原细菌的检测更快速、灵敏和可靠。本文对近年来植物病原细菌分子检测技术进行介绍,尤其是应用广泛的ITS-PCR(intergenictranscribedspace-PCR)、ARDRA(amplifiedribosomalDNArestric-tionanalysis-PCR)、rep-PCR(repetitiveDNA-PCR)和实时荧光定量PCR(real-timequantitativePCR)技术,旨在促进我国植物病原细菌研究的快速发展。 相似文献
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三种分子检测体系的比较及柑橘果园黄龙病监测 总被引：5，自引：5，他引：0
为了评价3种PCR分子检测体系对柑橘黄龙病(citrus huanglongbing,HLB)大田诊断效果,综合比较了常规PCR、巢式PCR和SYBR Green Ⅰ荧光定量PCR(SG Ⅰ-qPCR)方法对柑橘黄龙病菌检测的灵敏度、特异性和准确度等参数,并用SYBR Green Ⅰ荧光定量PCR和巢式PCR监测广西柑橘园疑似HLB样品425个,比较了2种检测体系的阳性检出率。基于CQULA04F/CQULA04R引物对的SYBR Green Ⅰ荧光定量PCR的灵敏度可达10 ag/μL;而巢式PCR灵敏度为100 ag/μL,巢式PCR较常规PCR检测灵敏度高104倍。疑似样品的HLB病原SYBR Green Ⅰ荧光定量PCR和巢式PCR检出率分别为46.6%、40.0%。各检测体系的灵敏性、特异性、符合度依次为SYBRGreen Ⅰ荧光定量PCR>巢式PCR>常规PCR。研究表明,SYBR Green Ⅰ荧光定量PCR可作为果园大规模HLB早期诊断和监测的首选,而在缺乏定量检测仪器时,巢式PCR也可用于HLB的检测,但需注意避免空气污染导致的假阳性。 相似文献
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根据番茄褪绿病毒(Tomato chlorosis virus, ToCV)热激蛋白70(Hsp70)的基因序列,设计ToCV实时荧光定量PCR特异引物。利用重组质粒ToCV-1为标准品建立SYBR Green I实时荧光定量方法。针对引物浓度、退火温度、特异性、灵敏度、重复性和稳定性进行系列优化。结果表明,最适退火温度为63℃,最适引物浓度为0.3 μmol·L-1。熔解曲线为特异性单峰,表明其特异性良好。建立的SYBR Green I实时荧光定量PCR较常规PCR灵敏100倍,且具有良好的重复性和稳定性。基于SYBR Green I实时荧光定量PCR技术建立的ToCV检测方法,速度快、特异性强、灵敏度高、重复性好,可以用于ToCV的定量检测。 相似文献
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Kelly Scarlett Len Tesoriero Rosalie Daniel David Guest 《European journal of plant pathology / European Foundation for Plant Pathology》2013,137(2):315-324
The rapid and reliable identification and quantification of pathogens is essential for the management of economically important plant diseases. Fusarium oxysporum f. sp. cucumerinum is the soil borne fungus responsible for Fusarium vascular wilt of cucumber. In this study, we report the development of a specific and reliable real-time quantitative PCR assay and the development of an ultra-sensitive diagnostic pseudo-nested PCR assay. The capacity of the PCR assays to accurately identify and quantify Fusarium oxysporum f. sp. cucumerinum was experimentally tested by the development of standard curves from serial dilutions of copy numbers in a range of complex environmental DNA samples. The amplification efficiency, sensitivity and reproducibility of the qPCR assays were not significantly affected by the presence of any of the non-target background DNA tested. In quantitative real-time PCR, as few as 100 copies could be reliably quantified, and in simple and pseudo-nested PCR as little as 10 pg and 10 fg, respectively, could be detected. This rapid and sensitive qPCR method can be used to facilitate investigations into plant–pathogen interactions, epidemiology, and disease management practices. 相似文献
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植物病害分子流行学概述 总被引：5，自引：1，他引：4
本文概要地介绍了植物病理学新研究领域——植物病害分子流行学的基本概念、进展和在几个主要方面的研究实例。分子生物技术在病原鉴定方面得到了广泛应用;定量的分子生物技术在测定病菌初侵染源方面显示出特有的快速、准确的优势;分子流行学应用分子生物技术监测病害和病原菌群体的动态,克服了传统流行学方法的弱点;作为有力的补充,分子生物技术正在用于探讨和推测病原菌远距离传播的路径,并注重研究病原菌群体的时、空动态变化,病原菌的长期进化,以及与病害发展的关系;病原群体的竞争将得到更深入的研究以揭示其变化是如何导致植物病害大流行;应用分子流行学手段,植物抗病性的鉴定将大大加速和简化;病害防治策略的制定将具有更科学的依据。宏观与微观研究手段的结合将越来越显示其在植物病害流行学研究中的巨大潜力。 相似文献
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科学施用杀菌剂是植物病害综合治理的重要措施之一, 然而由于杀菌剂的长期使用, 病菌抗药性问题逐渐加重, 严重影响药剂的防治效果和使用寿命。近年来, 随着分子生物学技术的快速发展, 人们对杀菌剂抗性机制有了更深入的理解, 并开发出了病菌抗药基因型快速检测的方法。本文总结了植物病原真菌对苯并咪唑类杀菌剂(BZD)、肌球蛋白合成抑制剂、甾醇脱甲基抑制剂(DMI)、QoI类抑制剂、琥珀酸脱氢酶抑制剂（SDHI）和二甲酰亚胺类杀菌剂(DC)的抗药性现状与抗性机制。在此基础上, 介绍了聚合酶链反应(PCR)、限制性片段长度多态性(RFLP)、等位基因特异性PCR和环介导等温扩增(LAMP)技术在杀菌剂抗性快速检测方面的研究进展。此外, 对抗药性治理对策进行了讨论和展望。 相似文献
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Quantitative plant resistance is supposed to be more durable than qualitative resistance for the control of plant diseases. However, it has been experimentally shown that erosion of quantitative resistance can occur. Cumulation of quantitative resistance loci (QRLs) in the same cultivar is considered to improve the efficiency and durability of quantitative resistance, but the choice of QRLs to be combined is of crucial importance. This study investigated whether the combination of QRLs acting on different stages of pathogen development could improve the efficiency of resistance in the apple scab pathosystem. The efficiencies of three QRLs were evaluated against 10 isolates of Venturia inaequalis and the stages of pathogen development that were affected by the QRLs were defined microscopically. A gain in the efficiency of resistance was observed when QRLs were pyramided compared to when they acted alone. Thanks to the combined effects of the individual QRLs, the pyramiding of the three QRLs hindered fungal development at different stages: before the penetration of the plant cuticule, after the penetration with hypersensitivity reaction, and during the colonization and asexual reproduction. These effects were dependent on the V. inaequalis isolates. These results suggest that the gain in efficiency of resistance by pyramiding may derive from the combination of different and complementary molecular mechanisms underlying QRLs. Thus, the resistance achieved from pyramiding such a combination of QRLs should be durable. 相似文献
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中国马铃薯晚疫病监测预警系统“China-blight”的组建及运行 总被引：4，自引：0，他引：4
马铃薯晚疫病是严重威胁世界马铃薯生产和粮食安全的重要病害之一,同时也是植物病害中流行速度最快的病害之一。由于品种多不抗病,目前国内外主要依靠化学防治控制该病害。为了提高用药的时效性,将信息技术与植物病害流行学原理相结合,设计并组建了中国马铃薯晚疫病监测预警系统"China-blight"(www.china-blight.net)。该系统由"中国晚疫病实时分布"、"未来48小时不同区域晚疫病菌侵染危险性预测"和"晚疫病化学防治决策支持系统"等子系统构成,此外还包括"晚疫病防治方法"、"品种抗病性"、"化学药剂库"、"其他病虫害"、"问题与经验交流"和"用户田间管理电子档案"等知识信息与服务功能。通过对2009年我国北方马铃薯一作区6-7月份病害侵染时段出现次数与晚疫病实际发生情况进行比较,预测信息与病害实际发生程度相符,该系统可以用于对马铃薯晚疫病田间防治的指导。 相似文献
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Grapevine leafroll‐associated virus 3 (GLRaV‐3) is associated with grapevine leafroll disease, one of the most economically important viral diseases of grapevines. This disease impacts on both vine health and grape quality; reduction in yield, brix and wine colour are among its detrimental effects. Many methods, including serological and molecular procedures, have been developed for the detection of GLRaV‐3; however, there is no PCR‐based assay available to quantify virus populations within plant tissues. A real‐time RT‐PCR assay with TaqMan probe was developed for specific and reliable quantitative detection of GLRaV‐3 in infected tissues. The designed primers and probes target the conserved sequence in the RNA‐dependent RNA polymerase (RdRp) domain of the viral genome to prevent amplification of most subgenomic and defective RNAs. This protocol was used to examine the seasonal dynamics and translocation of GLRaV‐3 in field‐grown grapevines. The results showed that the virus spread quickly from trunks to new growing shoots and leaves early in the growing season, and most samples still harboured detectable virus during late summer and autumn. The seasonal progress of one GLRaV‐3 isolate was compared in four grapevine cultivars (Chardonnay, Cabernet Sauvignon, Italia and Thompson Seedless). Within cultivars there was little variability in the distribution and translocation of GLRaV‐3, except for in Thompson Seedless. This quantitative detection assay will be a valuable tool for GLRaV‐3 diagnosis, disease monitoring and population ecology studies. 相似文献
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This paper presents the results of long‐term monitoring of insecticide resistance in populations of agricultural pests in Russia. Over the last 45 years, resistance developments were recorded for 36 arthropod pest species in 11 agricultural crops and pastures in relation to nearly all commonly used plant protection products. Development of group, cross and multiple resistance has been revealed in populations of many economically important pests. Toxicological and phenotypical (for Colorado potato beetle) methods have been devised to monitor the development of pesticide resistance. Based on experience over the last century, systems aimed at preventing the development of pest resistance to insecticides and acaricides are elaborated. These systems are based on resistance monitoring and using plant protection measures which minimize the toxic pressure on agroecosystems. 相似文献
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[目的]从黄龙病耐病寄主植物cDNA中筛选抗病基因相关序列并对其进行表达分析研究。[方法]根据已克隆的植物抗性基因表达产物NBS-LRR保守区域设计简并引物,以耐HLB的柑橘属柚cDNA为模板扩增RGAs,并进行实时荧光定量PCR。[结果]通过RFLP分析及克隆测序共得到5个NBS类抗病基因相似序列(RGAs)片段,在GenBank上登录号为HM777043～HM777047。通过Clustalx、DNAMAN等软件分析5个RGAs及其推导的氨基酸的相似性,结果显示它们均含有典型NBS-LRR类抗性基因所具有的保守区域:P-loop、Kinase-2a、GL-PLAL,其与已克隆的烟草N、亚麻L6、拟南芥RPS2、RPS5、RPP8、RPM1等抗病基因在保守区域氨基酸水平上的相似性为19.71%～42.86%。根据得到的序列设计特异性引物,对5个RGAs在HLB侵染过程中的表达进行定量PCR,结果显示嫁接病芽接穗后的8次连续采样中5个RGA的表达受到不同程度的调控。[结论]表明5个RGAs可能与黄龙病的侵染有关。 相似文献